ISSN 1110-6107 Ass. Univ. Bull. Environ. Res. Assiut, Egypt Vol. 14 No. 2, October 2011

An Environmental Journal Concerned With The Interaction Between Living Systems And Environment

EDITORIAL BOARD

Editor in Chief Prof. Dr. Ahmed A. Geies Vice President of Assiut University for Community Services and Environmental Affairs

Editorial Secretary Prof. Dr. Mohamed A-K. Mohamed Faculty of Engineering - Assiut University

Editorial Members

Director of AUCES Prof. Dr. Thabet A. Ibrahim Faculty of Vet. Med. - Assiut University

Prof. Dr. Fawzy M. Salama Prof. Dr. Ali H. Zarzour Faculty of Science - Assiut University Faculty of Medicine - Assiut University

Prof. Dr. Ahmed Ghallab Prof. Dr. Taha A. El-mestekawy Faculty of Agriculture - Assiut Faculty of Arts - Assiut University University

Prof. Dr. Abdel-aziz A. Fayed Prof. Dr. Hosam El-Din M. Omar

Faculty of Science - Assiut University Faculty of Science - Assiut University

Dr. - Ing. Adel Abdou H. Ahmed Faculty of Engineering - Assiut University ETCA Coordinator

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

EFFECT OF THE COLOUR OF FEEDERS AND DRINKERS ON DEVELOPMENT OF SOME MAINTENANCE BEHAVIORS IN NEWLY HATCHED CHICKEN Motamed Elsayed Mahmoud**, Reem Mahmoud Dosoky* and Moustafa Mohamed Ahmed* *Department of Animal Hygiene, Faculty of Veterinary Medicine, Assiut University, Egypt **Department of Animal Husbandry, Faculty of Veterinary Medicine, Sohag University, Egypt

ABSTRACT: Background: Colors of long wave lengths and color contrast can increase the responsiveness of chicken to stimuli. The purpose: This study was designated to assess the effect of feeders and drinkers colors and their contrast on ingestion rate and development of some maintenance behaviors Methods: This study includes 2 experiments. Experiment I: Effect of different colors of feeders and drinkers on rates feeding rates and water intake during 6 weeks rearing period was examined in Ross broiler chicken. Experiment II: Assessment of the effect of color contrast of feeders and drinkers with the floor on development of maintenance behaviors in newly hatched chicken. Results: The results indicated that red colored feeders and drinkers increased the feed and water intake rates during the first week post hatching (P<0.05). But no such effect was observed in older ages. In addition, the presence of color contrast feeders and drinkers with the floor enhanced the appearance of certain behaviors necessary for survival in newly hatched chicks. In conclusion red colored feeders and drinkers attracted chicks, increased ingestion rate and color contrast accelerated the development of some maintenance behaviors and decreased distress calls. Thus using red colors and presence of contrast environment could enhance the development of maintenance behaviors in newly hatched chicken.

INTRODUCTION: lights and yellow colors (Thomast & Lyons, 1968; Schaefer & Hess, 1969; Smith & Bird Chicken relied mostly on its sense of vision 1971 and Miklo´si et al. (2002). Also chicken for eating, drinking or even survival. Chicken preferred high color contrast or differing colors preferred colors of long wave lengths over that over familiar colors (Osorio et al., 1999, 2009). of short wave lengths. e.g. the responses of When chicks exposed to a choice situation and chicks to distant flashing lights were poor to the order of preference to colours recorded, the white and blue colors in comparison to red values from highest to lowest were as follows:

-1- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

blue, red, green and white respectively (Hess, maintenance behaviors in newly hatched chicks. 1959). In a preference test if the bird was given The present study was performed to know the a choice of certain aspects of its environment, it best desirable colors to suffice welfare and would choose according to how it felt. i.e. in the enhance productivity in chicken. best interest of its wellbeing or welfare (Duncan, MATERIAL AND METHODS: 1991). Subjects and housing: Chicken behavior, as the way in which it The study was performed in an acts, covers many activities or many patterns. experimental farm run by Animal and Poultry Feeding, drinking, body care and comfort are Production Department, Faculty of Agriculture, examples of maintenance behavioral patterns Assiut University, Egypt. All chicks were fed the (Broom, 1992). Other patterns of behavior same total mixed ration. Ambient temperatures concerned with conveying information to other were scheduled in a descending gradient (37 to birds of the same species are called displays 24°C) and lighting artificially provided by (Duncan & Wood-Gush, 1972 and Appleby et al. incandescent bulbs (1-5 watt/m2) with (hours; 1992). The process of food ingestion in birds was 23.5 L/0.5 D) daily light program (Rose, 1997). divided into 5 stages: arousal and food Vaccinations and medications were scheduled as recognition, orientation, grasping, recommended by manufacturer companies. mandibulation and swallowing (Kuenzel, 1983). Chicks were kept under veterinary supervision, Birds did not take more time when drink than selectively chosen and clinically healthy. did as they feed. In addition, there were no strain differences in drinking behavior. Experiment I: However, feeding and picking behaviors were Measuring effect of feeders and drinkers directly facilitated by social stimulation e.g. the colors on ingestive behavior in broiler presence of an active companion especially chicken during 6 weeks rearing period: during feeding. Birds comfort behaviors or It was performed on 40-Ross broiler chicks activities have to do with their body care. eg. from one day to 42-days old. Feeders and preening, dust bathing, scratching, head drinkers colors were red, green, white and blue. shaking, leg and wing stretching…etc. were an Locations of feeder and drinkers were oriented important part of the skin hygiene and self- from time to time to preclude the location maintaining behaviors (Fraser & Broom, 1990 preference (Duncan, 1991). In addition, the size and Appleby et al. 1992). of feeders and drinkers were increased In this study 2 separate experiments were gradually to adapt chicken ingestion rate. The performed to assess the effect of colors of following 2 rules of Duncan (1991) were adapted drinkers and feeders on feeding and drinking to avoid preference testing shortcomings: 1-A behaviors in chicken and development of wide rang of choices were made to avoid the

-2- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

only two percentage values results (eg: If the bird's neck was fully recumbent and the 70%+30%). 2-Limiting the test time to one hour eyes permanently closed, it was recorded as to assure the resulted preference value indicated sleeping (Vestegaard, 1982 and Broom, 1992). the current feeling of birds and allowed a Also any of walking, drinking, floor picking, sufficient time for birds to perform their full picking other birds, wing stretching, leg repertoire of behavior. Ingestion rate was stretching, preening…etc. or any other activities measured according to Murphy & Preston were recorded. In addition the frequencies of (1988) protocol with slight modifications; the such behaviors during the observation were consumption rate/chick/feeder or drinker/hour tabulated (Prayitno et al., 1997). were calculated by computing weighing Statistical analysis: differences among feeders/drinker full of All statistical analysis of data was ration/water (A), weighing of feeders/drinker performed using SPSS (2007) Software. All after test (B) and weighing of ration/water on values were presented as means±standard error the sheet underneath (C). (SEM). Groups at different ages were analyzed Experiment II: by two-way analysis of variance (ANOVA). Effect of colors contrast on development Tukey’s HSD was used for multiple of maintenance behaviors in newly comparisons among mean values in different groups. hatched chicks: The number of birds was 30 newly hatched chicks which are divided into 5 Total average feed and water intake were groups (6 chicks each). Each group was stocked calculated. Data are presented as mean±SE. in a plastic hatching basket (95×37×23 cm3) for Values with asterisks mean significant versus # one week. Each group of chicks were daily (P<0.05). observed for 10-min sessions and sequentially Data in Fig. 1 indicated that red colored repeated for 3- hrs (9:00-12:00) in the morning feeders and drinkers increased total feed and and 3-hrs (3:00-6:00) afternoon. Each chick water intake rate when compared with blue displayed a certain behavior was described and colored feeders and drinkers during first week. recorded as first time record to that behavior. Murphy and Preston (1988) found that mean e g.: feeding from dish, drinking, preening, dust values for feeders visit times were generally bathing, and perching, picking, vocalization, short; almost less than a minute. On contrast, resting, sleeping and dozing. Dozing was the drinker visit times were shorter than feeder recorded when the chicken was sitting with neck visit times. About 93% of all drinker visit times withdrawn, the head motionless and the eyes were less than 1 minute in length. The number either half closed or slowly opening and closing. of beak dips/time ranged from 1 to 27.

-3- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 1: Groups and types of feeders, drinker and floor contrast in plastic hatching baskets

Groups Name Feeder color Drinker color Floor color

No contrast (NC) Transparent Transparent White RF- RD Red Red White RF-NCD Red Transparent White NCF-RD Transparent Red White WF-WD White White Red

RESULTS & DISCUSSION: A B

14 * 14

g/hr)g/hr)g/hr)

( ( (

ml/hr)ml/hr)ml/hr)

ml/hr) 12

( ( ( 12 # ( * 10 10 #

8 8 6 6 4 4

2 2

Feed rate rate intake intake Feed Feed

Feed rate intake Feed

Water intake rateraterate intake intake intake Water Water Water Water intake rate intake Water 0 0 Red Green White Blue Red Green White Blue

Feeder color Drinker color

Fig. 1 : Effect of feeders and drinkers color on total feed (A) and water intake (B) rates (chick/hr) during 6 weeks period in Ross broiler chicken

Table 2: Effect of feeders and drinkers colours on ingestive behavior in Hubbard broiler chicken Age Feeder Color Drinker Color (day) Red Green White Blue Red Green White Blue 1st 5.6 ±0.2a 3.1 ±0.1 4.7 ±0.1 3.2 ±0.1 6.7 ±0.2 a 5.5 ±0.4 5.0 ±0.3 5.1 ±0.4 2nd 6.5 ±0.3 a 3.3 ±0.1 5.6 ±0.1 4.3 ±0.4 6.6 ±0.2 6.0 ±0.1 5.5 ±0.2 6.0 ±0.3 3rd 6.5 ±0.4 a 4.8 ±0.3 4.5 ±0.2 5.1 ± 0.2 8.8 ±0.4 a 6.5 ±0.3 5.4 ±0.4 5.6 ±0.1 4th 6.7 ±0.3 a 5.9 ±0.2 4.2 ±0.3 5.4 ±0.3 9.1 ±0.3 a 7.0 ±0.3 b 5.1 ±0.5 5.5 ±0.2 5th 7.1 ±0.3 a 4.5 ±0.3 4.9 ±0.4 5.1 ±0.4 8.6 ±0.5 a 7.6 ±0.1 b 5.6 ±0.2 6.1 ±0.4 6th 6.9 ±0.4 a 4.8 ±0.2 4.9 ±0.1 5.8 ±0.3 8.7 ±0.3 a 6.1 ±0.3 5.1 ±0.3 6.4 ±0.5 7th 7.6 ±0.5 7.2 ±0.3 6.9 ±0.5 6.9 ±0.5 8.0 ±0.2 5.0 ±0.2 5.7 ±0.5 5.2 ±0.3 8th 7.3 ±0.2 a 6.5 ±0.4 b 5.8 ±0.3 5.9 ±0.3 7.7 ±0.3 7.1 ±0.4 8.5 ±0.3 7.9 ±0.2 9th 8.1 ±0.3 7.5 ±0.3 7.8 ±0.4 7.6 ±0.4 7.8 ±0.2 7.8 ±0.5 8.4 ±0.4 7.9 ±0.5 10th 8.0 ±0.4 7.7 ±0.5 7.9 ±0.3 7.5 ±0.3 8.6 ±0.3 7.6 ±0.6 8.3 ±0.3 8.2 ±0.2

Feed intake rate (chick/gram/hr) from advancing age. Tukey’s HSD tests indicated that feeders and water intake (ml/hr) from drinkers red feeder and drinker increased the feed intake during first 10 days post hatching were rates in chicks up to 6 days post hatching. Data measured. Two ways ANOVA indicated an age are presented as mean±SE. Values with and color effect on feed and water intake. There superscripted letters mean significant (P<0.05) was a general increase in ingestion rate with differences.

-4- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Results in Table 2 could be interpreted by (preening and stretching), resting and sleeping finding of Gray (1961) who mentioned that the and vocalization.

responses of broiler chicks to colours were high Since the time of the day had no significant to red color when compared with white, blue effect on the following behaviors; wing/leg and green colors during first 5 days post- stretch, wing flapping, head scratching and hatching. Regarding the effect of age on preening (Nicol, 1987 a & b) this finding could ingestion Richard et al. (1997) and Savory & validate time limited observation schedule in the Mann (1997) observed an age-related increase present study. in ingestive behavior because the frequency of With respect to feeding behavior, NCF- its corresponding exploratory behavior group and also WF-WD group only one chick decreased with advancing age in chicken. identified and reached feeder at first day. While However Newberry et al. (1988) and Blokhuis & all chicks developed feeding within the first 3 Haar (1992) found that frequency of feeding hours after provision o feeders and drinkers in and drinking declined as the age of chicks other groups. RF-WD- group developed feeding increased. pattern during the first 90-min of observation Effect of color contrast of feeders and all chicks. This may be attributed to the good drinkers on appearance and frequency of contrast made by red feeder that could be very behaviors in newly hatched chicks during 3 attractive for chicks to begin picking. All chicks hours daily observation. Behaviors recorded of WF-RD group reached the feed dishes and were counted/ group/3 hours observation. Key: showed picking and feeding during the first 45- NC; No contrast by colorless feeders and minutes. WF-WD-group with red colored floor drinker, RF- RD; red feeder and red drinker, possibly made the best contrast to the white RF-NCD red feeder and colorless drinker, NCF- colored feeders and yellowish colored ration. On RD; colorless feeder and red drinker, WF-WD; other hand, all chicks in NC-group appeared white feeder and white drinker. Values scale: lazy and drowsy and remained without (+=1-5, ++= 6-10, +++= 11-15 and ++++ = over noticeable movement during the first day 15). observation hours. Only one chick reached the First of all, behaviors observed during first feeder. Colourless feeder had the worst contrast week post hatching were nutritional (feeding, with the white floor and probably was not drinking, and elimination), body care behaviors attractive to newly hatched chicks.

-5- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 3: Qualitative assessment of the effect of color contrast of feeders, drinkers and floor on appearance and frequency of behaviors in newly hatched chicks Group Behavioral pattern recorded first NC RF-RD RF-NCD NCF- RD WF-WD First Day Feeding bouts ++ ++++ +++ + + Drinking bouts in form of pick drinking ++ +++ ++ ++++ ++++ Elimination times + + + + + Picking of grains from beaks of other birds + + - + + Ground picking - - - - + Sleeping with head on ground + + + + + Sleeping with beak on ground + + + + + Loud vocalization calls ++ + + +++ + None loud vocalization calls + + + + + AutoPreening + ++ + + ++ AlloPreening + + - - + Resting + - + - ++ Dozing + ++ + - ++ Second Day Toe picking + - - + - Ground Picking + - - - - Leg stretching + + + - + Wing stretching - - + - + Third Day Drinking bouts in form of scope drinking + +++ +++ ++ ++ Drinking bouts in form of pick drinking + - + - + Autopreening of back while standing ++ ++ ++ ++ +++ Autopreening of outside wing while standing + + + + ++ Autopreening of inside wing while standing - - - - + Autopreening of wing while sitting + + + ++ + Autopreening of back while sitting + + + + + Picking of environmental conspecifics - + + + + Wing flapping + - - + - Fourth Day Drinking bouts in form of pick drinking + + + + + Drinking bouts in form of scope drinking + + + + + Autopreening of head by leg while standing + + + + + Floor scratching + + + + - Perching on feeder + + + + - Fifth Day Scoop drinking + + + - + picking drinking + + + + + Dust bathing in feed trough + ++ ++ - + Six Day Scope drinking + + + + + Autopreening of back + - - + + Stretching of wing and leg + + + + ++ Seventh Day Autopreening of wing while sitting + - + + - + = 1-5, ++ = 6-10, +++ = 11-15 ++++ = over 15.

-6- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Regarding drinking Behavior, initially, it highest picking to environmental conspecifics. was observed that all chicks performed a less With respect to the NC-group the absence of developed form of drinking that was not like the any environmental contrast or any color ordinary scope form of drinking. This form of attractive in their neighborhoods perhaps was drinking continued up to the 3rd day post the causal factor for increased a searching for hatching. Our finding in present study was that food component of exploratory behavior or as head movement during drinking was the only redirected ground picking (Broom, 1992). The component similar to developed scope form of latter probably indirectly reflected in increased drinking. In WF-RD-group was the first group conspecifics' picking. Picking other birds or showed drinking. All chicks attracted to the cannibalism was not observed in the present drinker tried to investigate water and began to study. This may be related to elimination of this drink within the first 5 min after placing the trait by selective breeding because nervousness drinker dish in the plastic hatching basket. This was found to be negatively correlated to meat may be due to the red colored drinker was the production in chicken (Jones, 1996). only attractive thing inside chicks environment. According to Fraser & Broom (1990) On contrast, chicks of the NC-group were the preening can divided into 2 categories: auto- last group reached drinkers. It was after about preening or itself body care and allo-preening 15-20- minutes. The remaining 3 groups, within or their selves body care. Preening can also 5-to 10-minutes began drinking. During the divided according to the part of the body third day all chicks from all groups showed engaged in preening activity into wing, body developed form of scope drinking. and head body care or preening behaviors. In Considering picking behavior, day ground present study body care behaviors, preening, or picking was firstly appeared in the WF-WD combing the feather covering the body appeared group during the first. That could be related to firstly by combing the back region. Back attraction by red floor. During the second day, preening was observed in all groups within the picking of environmental conspecifics observed first three hours post hatching. In one day old in all groups of birds with varied frequencies. chicks, head preening activity was only Hence all chicks at second day showed feeding observed in the NCF-RD group. activity picking of that could be initiated by The highest allopreening activity was feeding. This result agreed with Fischer, et al. observed in the WF-WD group that probably (1975) presumed that, newly hatched chicks did correlated to the highest action of ground not pick much more before their second day of picking that may be considered as redirected life because of the compensated utilization of the ground picking behavior. On other hand NC- yolk sac during such period. During the third group did not show any allo-preening activity. day, NCF-RD and WF-WD-groups showed the These results could be attributed to hunger

-7- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

associated with inactivity. However, Webster on the floor. Ground scratching considered as a (2000) and Na¨tt, et al. (2007) found that component of dust-bathing behavior (Fraser stimulations of preening by feather growth and Broom, 1990). But Blokhuis & Harr (1992) might the most important causal factor for the observed that ground picking and scratching development of that behavior. were more common after feeding period (s) than Regarding sleep, huddling and sleeping was before. In present study chicks 5th day old observed in two groups; NC-group and WF-RD showed dust bathing in feeders this result group during the first day. Chicks of these agreed with Murphy & Preston (1988) and groups were apparently inactive remained Broom (1992) who observed that chickens motionless and walked in lazy manner during would dust-bathe if sand or other alternative first day. To elucidate the appearance of materials were available. If there were no huddling and sleeping in these 2 groups could be alternatives chickens would dust-bathe in dry due to lowered feed visits and consequent food. Vestegaard (1982) recorded the decreased feed consumption and consequent development or appearance of dust-bathing diminished heat increment. After that birds felt behavior in chicks when they were about few cold and tried to warm that could be found by days post hatching. On other hand Murphy & huddling together. Allam (1994) mentioned that Preston (1988) and Savory & Mann (1997) huddling provided warmth function by contact interpreted the lower frequency of dust bathing, and decreased heat loss. preening and non-damaging picking in broiler In the present study loud and non loud chicken due to the intensive selection against vocalizations were emitted by chicks from all these behavioral traits in broilers as these traits groups. During the first day the variations was were undesirable from economic point of view. as following: Chicks of NC-RD group produced Vestegaard. (1982) considered allopreening or the highest frequency of vocalization of loud allopicking to be the result of an abnormal type (17- calls) followed by NC-group. Chicks of development of the perceptual mechanisms this group showed the lowest feeding an4d responsible for detection of some substrates, drinking bouts during first day (Table 3). These these substrate materials were needed for calls could be attributed to thirst, hunger or performance of dust-bathing behavior. In other distress calls. In agreement with that Andrew words, they explained allopreening as redirected (1973) who named loud vocalization calls by behaviors of another behavioral pattern hunger or distress calls. (ground picking). Regarding ground scratching, chicks of In conclusion, to conclude red colored WF-WD-group were the only group showed feeders and drinkers attracted chicks, increased scratching ground. Chicks scratched the floor ingestion rate and color contrast accelerated the with their legs and sprayed the loose materials development of some maintenance behaviors

-8- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

and decreased distress calls. Thus using red Fraser, A.F. and Broom, D.M. (1990): Farm colors and presence of contrast environment Animal Behavior & Welfare. Third could enhance the development of maintenance edition, chapter: 10 pp: 90-110. behaviors in newly hatched chicken. Gray, P.H. (1961): The releasers of imprinting: differential reactions to colours as a REFERENCES: function of maturation. Journal of

Allam, S. (1994): Poultry management and comparative physiology and psychology. breeding. Chapter 7. PP.169-180. 5: 597-601. Andrew, R.J. (1973): Vocalization in chicks, Hess, F.H. (1959): Imprinting in broiler chicks. And the concept of “stimulus contrast” Science, 130 (3368): 133-141. Animal Behavior. 23: 64-76. Jones, R.B. (1996): Fear and adaptability in Appleby, M.C.; Hghes, B.O. and Elson, H.A. poultry: insights, implications and (1992): Poultry production systems. imperatives. World’s Poult. Sci. J. 52, Behavior, Management and Welfare. 131-174. C.A.B. International. Kuenzel, W.J. (1983): Behavioral sequences of Blokhuis, H.J. and Haar, V. (1992): Effect of food and water intake: its significance for picking incentives during rearing on elucidating central neural mechanisms feather picking of laying hens. Br. Poult. controlling feeding in birds. Bird Sci. 33 (1): 17-24. Behavior 5:2-15. Broom, D.M. (1992): The needs of laying hens Miklo´si, A ´, Gonda, Zs., Osorio, D. and Farzin and some indicators of poor welfare in A. (2002): The effects of the visual the laying hen. British veterinary environment on responses to colour by Journal, 142: 524-526. domestic chicks J Comp Physiol. A 188: Duncan, J.H. (1991): Measuring preferences 135–140. and strength of preferences. Poultry Murphy, L.B. and Preston, A.P. (1988): Food Science. 71: 658-663. availability and the feeding and drinking Duncan, I.J. and Wood-Gush, D.G. (1972): An behavior of broiler chickens grown analysis of side placement preening in commercially. British poultry science. 29: domestic fowl. Animal Behavior. 273-283. 20: 68-71. Na¨tt, A., Kerje, E., Andersson, L. and Jensen, Fischer, A.E.; Morris, G.L. and Ruhson, D.E. P.(2007): Plumage color and feather (1975): Colour picking preference in pecking-behavioral differences associated white leghorn chicks. J. of comparative with PMEL17 Genotypes in Chicken physiology and psychology. 88: 402-406. (Gallus gallus). Behav Genet 37:399-407.

-9- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Newberry, R.C.; Hunt, J.R. and Cordiner, EE. Revate d.Elevage et de Medecine (1988): Influence of light intensity on vétérinaire des pay tropicaux 50: (1) 65- behavior and performance of broiler 74. chickens. Poult. Sci., 67: 1020-1025. Rose, S.P. (1997): Principles of poultry science. Nicol, C.J. (1987a): The effect of cage height on Chapter g: PP. 117-128. C.A.B. Interna- the behavior of hens housed in battery tional Wallingford. Oxon. U.K. cages. Poultry Science 28: 327-335. Savory, C.J & Mann, J.S. (1997): Behavioral Nicol, C.J. (1987b): Behavioral responses of development in groups of pen houses laying hens following a period of spatial pullets in relation to genetic strain, age restriction. Animal Behavior 35: 1709- and food form. Br. Poult. Sci. 38: 38-47. 1719. Schaefer, H.H. and Hess, E.H. (1969): Colour Osorio, D., Vorobyev, M., and Jones, C.D. preferences in imprinting objects. Z. (1999): Color vision of domestic chicks. Tierpsychol., 16, 2, 161-172. The Journal of Experimental Biology, Smith, F.V. and Bird, M.W. (1971): The 202, 2951–2959. sustained approach of the domestic chick Osorio, D., Ham, A.D., Gonda, Z. and Andrew, to coloured stimuli. Animal Behavior. 20: R.J. (2009): Sensory generalization and 60-63. learning about novel colours by poultry SPSS (2007): Statistical Package for Social chicks. Q J Exp Psychol (Colchester). Science, version 16 (2007) for Window. 62(7):1249-56. Thomast, D.R. and Lyons, J. (1968): Visual field Prayitno, D.S.; Phillips, C.J.; and Stokes, D.K. dependency in pigean. Animal behavior (1997): The effect of colour and intensity 16, 213-218. of light on Behavior and leg disorders in Vestegaard, K.J. (1982): Dust bathing in the broiler chickens. Poultry science. 76: domestic fowl. Diurnal rhythm and dust 1674-1681. deprivation. Applied Animal Ethology. 8: Richard, P.; Vilarino, M.; Faure.; Leon J. and 487-496. Picard, M. (1997): Study of broiler Webster, A.B. (2000): Behavior of white leghorn chicken behavior in tropical intensive laying hens after withdrawal of feed. breeding poultry house in venezuela. Poultry Science 79: 192-199.

-10- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

تأثير لون الغذايات والمشربيات على تطور بعض سلوكيات البقاء في كتاكيت الدجاج حديثة الفقس معتمد السيد محمود، ريم محمود دسوقي، مصطفى محمد أحمد

الخلفية البحثية: التباين في األلوان داخل نظام التربية المكثفة حيث تووافر اضاوا ا المنخفاوة واألدوداد الكبيورا ثود يو ثر دلووا اجووت ابة الوود اث للموو ثراي الخار يووة الغوورم موون الدراجووة: هووو دراجووة تووعثير بوووم األلوووان المختلفووة للغوو اياي والمشووربياي مثوول األلوووان اي األلوووال المو يووة ال وويرا )األزرق واألخاوور( واللويلووة )األحموور( واللووون األبوويم دلووا الجلوك الغ ائي باضاافة إلا دراجوة توعثير تبواين األلووان اي ااجوت ابة الواليوة دلوا تلوور بووم جولوكياي الب وا فوي كتاكيي الد اث حديثة الف س. لرق البحث والنتائج: تم إ را ه ه الدراجة دلا ت ربتين. الت ربوة األولوا: ل يواس توعثير األلوان المختل ة )األحمر، األخاور، األبويم واألزرق( لكول مون الغو اياي والمشوربياي دلوا موودل تنواول الغو ا والشور لكتاكيي د اث الروس كمثال لبدارى التجمين الت ارية، أواحي ه ه الدراجوة أن الغو اياي والمشوربياي الحمورا أدي الوا زيادا مودل تنواول الغو ا والشور فوي األجوبول األول مون الومور ولكون لوم يححوظ هو ا التوعثير فوي الومور األكبور. الت ربوة الثانية ل ياس تعثير تبواين األلووان اي ااجوت ابة المرتفووة )األحمور( للغو اياي والمشوربياي موأل األراوية كخلفيوة للتبواين دلا ظهور بوم جلوكياي الب ا في الكتاكيي حديثة الف س. وتبين من النتائج أن و ود تباين األلوان في بيئة كتاكيوي الد اث أدى الا اضجرال من تناول الغ ا ، وك لك ظهور مبكر لبوم جلوكياي الب ا دند م ارنتها بالبيئة التي ا يو د بها تباين. ااجتنتاث: اجت ابة الكتاكيي للون األحمر والتباين في األلووان يكوون أدلوا فوي الكتاكيوي حديثوة الف وس دنو في الومر األكبور مموا ي ودر بال وائمين دلوا ردايوة الود اث ااهتموام بوعلوان الغو اياي والمشوربياي تواي واد التبواين لهوا فوي المووزارل خ و ووا الكتاكيووي حديثووة الف ووس األموور الوو ع يوونوكس بوودوره دلووا ااجووت ابة للغوودا والشوور وبالتووالي دلووا اضنتا ية.

-11- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

ROLES OF MICROALGAE AND BACTERIA IN HYDROGEN PRODUCTION AS ONE OF THE RENEWABLE ENERGY RESOURCES Ahmed M.A. Abdel-Monem* and Farag M. A. Shaieb

Botany Department, Faculty of science, Omar Al-Mukhtar University, Al-Beida, Libya, P.O. 919, Fax. (+218) 84637052. Tel. (+218) 84 632946 – 84 632233 *Correspondence author: E. mail: [email protected] , mob. (+218) 927884159 Currant address: 2 Nadi Algomhourea St., Shbeen El-Koom, El-Menoufia, Mob: 010 3066695

REVIEW ARTICLE:

ABSTRACT: Hydrogen gas is considered to be one of the most desired alternate sources of the limited fossil energy resources of today. It shows great promise as a non-polluting fuel, but to reduce carbon dioxide releases hydrogen gas will need to be produced from renewable sources. The limited fossil fuel prompts the prospecting of various unconventional energy sources to take over the traditional fossil fuel energy source. Photosynthetic microbes can produce hydrogen using the nature plentiful resources, sunlight, the included greens, and blue-green algae (Cyanobacteria), either via direct or indirect biophotolysis. In addition, Cyanobacteria produced hydrogen through decomposing the organic compounds (Photodecomposition). The hydrogen production by green algae could be considered as an

economical and sustainable method, water utilization as a renewable resource and recycling CO2, a greenhouse gas. Rates of hydrogen production by photoheterotrophic bacteria are higher in the case of immobilized cells than that of the suspended cells. Cyanobacteria are highly promising microorganism for hydrogen production. Cyanobacterial hydrogen production is commercially viable, in comparison to the traditional ways of hydrogen production (chemical, photoelectrical). The present review shows the basic biology of microalgae and bacterial hydrogen production and its future prospects. While integrating the existing knowledge and technology, much future improvement and progress is to be done before hydrogen is accepted as a commercial primary energy source.

INTRODUCTION: green house gases (i.e., methane and carbon dioxide), developing clean and new energy will Hydrogen gas shows great promise as a non- be one of the important researches in the polluting fuel, but to reduce carbon dioxide further. Hydrogen, high energy yield (about releases hydrogen gas will need to be produced 2.75 times greater than that of hydrocarbon from renewable sources. Due to the fuels), is considered a promising candidate as an consumption of fossil fuel and production of

-153- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

ideal and clean source of energy. Biohydrogen Hydrogen has the highest energy content production process not only can solve per unit weight of any known fuel and can be environmental pollution, but also achieve transported for domestic/industrial resource recycling (Chang, 2001). consumption through conventional means. H2

Hydrogen (H2) offers tremendous potential gas is safer to handle than domestic natural gas. as a clean, renewable energy currency. H2 is now universally accepted as an Hydrogen has the highest gravimetric energy environmentally safe, renewable energy density of any known fuel and is compatible resource and an ideal alternative to fossil fuels with electrochemical and combustion processes that doesn’t contribute to the greenhouse effect. for energy conversion without producing The only carbon-free fuel, H2 upon oxidation carbon-based emissions that contribute to produces water alone. H2 can be used either as environmental pollution and climate change. the fuel for direct combustion in an internal Hydrogen fuel cells and related hydrogen combustion engine or as the fuel for a fuel cell. technologies provide the essential link between The largest users of H2, however, are the renewable energy sources and sustainable fertilizer and petroleum industries with, energy services. The transition from a fossil respectively, 50% and 37% (Momirlan and fuel-based economy to a hydrogen energy-based Veziroglu, 2002). Presently, hydrogen is economy, however, is fraught (Dunn, 2002). produced 40% from natural gas, 30% from It can fill an important role in the heavy oils and Naphtha, 18% from coal, and "greening" of the global energy and industrial 4% from electrolysis (Nath and Das, 2003). base. H2 is not a greenhouse gas, it has 2.4 times Hydrogen intensive research work has the energy content of methane (mass basis) and already been carried out on the advancement of its reaction with oxygen in fuel cells produces these processes, such as the development of only harmless water. Not only can pollutants genetically modified microorganism, metabolic from fuels used in high-temperature combustion engineering, improvement of the reactor engines be avoided using hydrogen-based fuel designs, use of different solid matrices for the cells, but the elimination of combustion also immobilization of whole cells, biochemical avoids the generation of NO2. As a result of assisted bioreactor, development of two-stage these advantages of hydrogen-based fuel cells, processes, etc. for higher H2-production rates there is a global transition occurring to (Debabrata and Veziroglu, 2008). Hydrogen hydrogen-based technologies. However, may be produced by a number of processes, hydrogen is currently produced mostly from including electrolysis of water, thermocatalytic fossil fuels, an inherently non-sustainable reformation of hydrogen-rich organic technology. compounds, and biological processes. Currently, hydrogen is produced, almost exclusively, by

-154- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

electrolysis of water or by steam reformation of wastes are available, thus avoiding energy methane. Biological production of hydrogen expenditure and costs for transport. In addition, (biohydrogen), using (micro) organisms, is an these are becoming important mainly due to exciting new area of technology development utilize renewable energy resources. These that offers the potential production of usable processes are usually carried out by different hydrogen from a variety of renewable resources. anaerobic bacteria and/or algae. The characteristics of these microorganisms widely However, major bottlenecks for the differ from each other with respect to substrates commercialization of these processes are lower and process conditions. The merits and demerits H2 yield and rate of H2 production. Suitable microbial cultures are required to handle waste of the processes have already been discussed (Das and Veziroglu, 2001, Nandi and Sengupta, materials efficiently, which are usually complex 1998) in nature. This will serve dual purposes: clean Biological hydrogen production processes energy generation and bioremediation. Scale-up offer a technique through which renewable studies on fermentative H2-production processes energy sources like biomass can be utilized for have been done successfully. Pilot plant trials of the generation of the cleanest energy carrier for the photo-fermentation processes require more attention. Use of cheaper raw materials and the use of mankind (Debabrata and Veziroglu, efficient biological hydrogen production 2008). Hydrogen has been produced through thermal cracking or water electrolysis, which processes will surely make them more requires much energy and emits global warming competitive with the conventional H2 generation processes in near future (Debabrata and gases such as carbon dioxide. One of the most promising methods is that the gas is produced Veziroglu, 2008). from annually renewed biomass using Biohydrogen has gained attention due to its microorganisms. potential as a sustainable alternative to the conventional methods for H2 production. It Microorganisms producing hydrogen: gives unassailable flexibility for a sustainable The types of microorganism-producing energy system, considering the present energy hydrogen are divided into four groups: crisis and environmental tribulations. Biological cyanobacteria, anaerobic bacteria and processes, unlike their chemical or photosynthetic bacteria. Cyanobacteria split electrochemical counterparts, are catalyzed by water into hydrogen and oxygen gas by microorganisms in an aqueous environment at photosynthesis. Anaerobic bacteria use organic ambient temperature and atmospheric pressure. substrates as the sole source of electrons and Furthermore, these techniques are well suited energy and convert them into hydrogen. The for decentralized energy production in small- reaction is rapid and the process does not scale installations in locations where biomass or require solar radiation, which makes it useful

-155- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

for treating large quantities of wastewater. using one of nature, most plentiful resources, Finally, the photosynthetic bacteria lie sunlight (Das and Veziroglu, 2001). somewhere between cyanobacteria and Hydrogen production rates of various anaerobic bacteria. Although photosynthetic biohydrogen systems are compared by first bacteria also convert organic substances to standardizing the units of hydrogen production hydrogen at fairly high rates, they also require and then by calculating the size of biohydrogen light energy to assist or promote the reactions systems that would be required to power proton involved in the hydrogen production. Some exchange membrane (PEM) fuel cells of various nonphotosynthetic bacteria can produce sizes. (Levin et al., 2004). hydrogen from different organic substrates, Generally; Biological hydrogen production such as Enterobacter Aerogenes from glucose or processes can be done by: a Clostridium beijerinckii from glucose and  Biophotolysis; green algae and blue-green starch. Non-photosynthetic bacteria, like algae (Cyanobacteria) split water molecules Clostridium butyricum, produce hydrogen from into hydrogen ion and oxygen. carbohydrates at a high rate, but the yield is low  Photodecomposition photosynthetic bacteria because they also produce organic acids. decompose the organic compounds by; Dark Photosynthetic bacteria such as Rhodobacter fermentation; and Hybrid systems. sphaeroides RV are powerful hydrogen producers, showing 7% energy conversion Direct biophotolysis: efficiency in the presence of lactate and Green algae, under anaerobic conditions, glutamate. The high rate of hydrogen can either use H2 as an electron donor in the production makes them suitable for CO2-fixation process or evolve H2. Hydrogen photobioreactor applications (Jeong et al., production by green microalgae requires 2008). several minutes to a few hours of anaerobic Hydrogen Bioproduction pathways: incubation in the dark to induce the synthesis and/or activation of enzymes involved in H A major route for hydrogen production is 2 metabolism, including a reversible hydrogenase biological nitrogen fixation (Prince and enzyme. Kheshgi, 2005). This is catalyzed by the enzyme Conversion of water to hydrogen by green nitrogenase, and hydrogen is an obligatory, but algae may be represented by the following not advantageous, product of a reaction that general reaction: evolved to enable cells to synthesize ammonia from nitrogen gas (Simpson and Burris, 1984). 2 H2O + light energy  2H2 + O2

So, Photosynthetic microbes can produce the The well-known H2-producing green algae, clean-burning fuel hydrogen gas (hydrogen) Chlamydomonas reinhardtii, under anaerobic

conditions, can either generate H2 or use H2 as

-156- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

an electron donor (Winkler, et al; 2002). The as a renewable resource and recycling CO2, a generated hydrogen ions are converted into greenhouse gas. However, strong inhibition hydrogen gas in the medium with electrons effect of generated oxygen on hydrogenase is the (donated by reduced ferredoxin) by major bottleneck for the process. It has been hydrogenase enzyme present in the cells. Light reported that inhibition of the hydrogenase by energy absorbed by photosystem II (PSII) oxygen can be partially overcome by cultivation generates electrons which are transferred to of algae under sulfur deprivation for 2–3 days to ferredoxin using light energy absorbed by provide anaerobic conditions under the light photosystem I (PSI). A reversible hydrogenase (Pinto et al., 2000). Major drawbacks of this accepts electrons directly from the reduced process are low hydrogen production potential ferredoxin to generate H2 in presence of and inability to use organic wastes. The hydrogenase. This enzyme is very sensitive to hydrogenase activity of C. reinhardtii [200

O2. Hydrogenase activity has also been observed nmol/(g chl a h)] is higher than Scenedesmus sp. in other green algae like Scenedesmus obliquus, [150 nmol/(g chl a h) (Winkler et al., 2002). Chlorella fusca (Winkler et al., 2002), Rates of hydrogen production by Chlorococcum littorale and Platymonas photoheterotrophic bacteria are higher in the subcordiformis (Nandi and Sengupta, 1998). On case of immobilized cells than that of the the other hand, there are several green algae suspended cells. types that do not have hydrogenase activity such as Dunaliella salina and Chlorella vulgaris Indirect biophotolysis: (Nandi and Sengupta, 1998). General reaction for hydrogen formation The hydrogen production by green algae from water by cyanobacteria (blue-green algae) could be considered as an economical and can be represented by following reactions: sustainable method, in terms of water utilization

12 H2O + 6CO2 + light energy  C6H12O6 + 6 O2 and

C6H12O6 + 12 H2O + light energy 12H2 + 6 CO2

Cyanobacteria are large and diverse group use is in the versatile metabolic capabilities of of photoautotrophic microorganism, contain these organisms and the lack of Photosystem II photosynthetic pigments such as Chl a, (PSII), which automatically eliminates the carotenoids and phycobiliproteins, and can difficulties associated with O2 inhibition of H2 perform oxygenic photosynthesis. Photosyn- production. thetic bacteria have long been studied for their Morphologically these organisms include capacity to produce significant amounts of unicellular, filamentous and colonial species. hydrogen (Bolton, 1996). The advantage of their Hydrogen is produced both by hydrogenase and

-157- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

nitrogenase enzymes. Within the filamentous The growth conditions for Anabaena are Cyanobacteria, vegetative cells may develop into simple which include nitrogen free media, structurally modified and functionally illumination, CO2 and N2. Nitrogenase plays specialized cells. The nutritional requirements important role for the hydrogen generation. of Cyanobacteria are simple: air (N2 and O2), Activity of the nitrogenase is inhibited by water, mineral salts and light. Hydrogen oxygen. Hydrogen production takes place under producing cyanobacteria may be either nitrogen anaerobic conditions. Some cultures require fixing or non-nitrogen fixing. The examples of CO2 during hydrogen evolution phase, although nitrogen fixing organisms are non-marine CO2 is reported to give some inhibition effects

Anabaena sp., marine Cyanobacteria Calothrix on photo-production of H2. Lower CO2 sp., Oscillatoria sp. Non-nitrogen fixing concentrations (4–18% w/v) have been reported organisms are Synechococcus sp., Gloebacter sp. to increase cell density during growth phase, and Anabaena sp. They are found suitable for resulting in higher hydrogen evolution in the higher hydrogen evolution as compared to other later stage (Liu et al., 2006). Simple sugars have cyanobacteria species (Pinto et al., 2002). been found suitable for hydrogen production. Heterocystous filamentous Anabaena cylindrica Recently more emphasis has been given to is a well-known hydrogen producing increase hydrogenase activity and bidirectional cyanobacterium. But, Anabaena variabilis has hydrogenase deficient mutants of Anabaena sp. received more attention in recent years, because to increase the rate of hydrogen production. of higher hydrogen yield (Liu et al., 2006). However, at the present time the rate of Hydrogen production by vegetative cells can hydrogen production by Anabaena sp. is take two routes: considerably lower than that obtained by dark A- Heterocystous nitrogen fixing bacteria. or photo-fermentations (Levin et al.; 2004). B-Nonheterocystous nitrogen fixing bacteria: With dinitrogen:

+ N2 + 8 H + 8 e_ + 16 ATP  2 NH3 + H2 + 16 ADP + 16 Pi Or, without dinitrogen + 8 H + 8 e_ + 16 ATP  4 H2 + 16 ADP + 16 Pi

A very wide variety of Cyanobacterial producing bacteria. That were concluded that species and strains has been studied for the aerobic bacteria will be used in the actual Hydrogen production. It occurs within at least system for hydrogen production from 14 Cyanobacteria genera, under a vast range of carbohydrate, while the anaerobic bacteria may culture conditions )Lopes et al., 2002(. be a good choice for the production of hydrogen Quantitatively, studies were investigated from wastewater containing innumerable between aerobic and anaerobic hydrogen- compound (Jeong et al., 2008). Some strains

-158- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

produced up to five times more hydrogen than bacteria have been found suitable to convert did wild-type cells growing under nitrogen- light energy into H2 using organic wastes as fixing conditions (Rey et al., 2007). They substrate (Liu et al., 2006), in batch processes revealed that in addition to the nitrogenase (Zurrer and Bachofen, 1979), continuous genes, 18 other genes are potentially required to cultures (Fascetti and Todini, 1995), or produce hydrogen. immobilized whole cell system using different Different microorganisms participate in the solid matrices like carrageenan (Francou and biological hydrogen generation system, green Vignais, 1984), agar gel (Vincenzini et al.,1986), algae, Cyanobacteria (or blue-green algae), porous glass (Tsygankov et al; 1994), and photosynthetic bacteria and fermentative polyurethane foam (Fedorov et al., 1998). bacteria were collected and illustrated in Certain photoheterotrophic bacteria within the Table (1). superfamily Rhodospirillaceae can grow in the

dark using CO as the sole carbon source to Photodecomposition of organic generate ATP with the simultaneous release of compounds by photosynthetic bacteria: H2 and CO2 (Winkler et al.,2002). The oxidation Phototrophic bacteria require organic or of CO to CO2 with the release of H2 occurs via a inorganic electron source to drive their water gas shift reaction as shown below: photosynthesis. They can utilize a wide range of CO + H2O → CO2 + H2 cheap compounds. These photoheterotrophic

Table 1: Reported hydrogen yield by different microorganisms (cited from; Debabrata and Veziroglu 2008)

Maximum rate of Maximum H2 yield Microorganisms Raw material used H2 production (mol H2/mol (ml/l.h) substrate) Green algae Scenedesmus obliquus TAP-S medium 3.6a – Chlamydomonas reinhardii TAP-S medium 4.5a – Chlamydomonas moewusii TAP-S medium 10.0a – Cyanobacteria Heterocystous Anabaena variabilis BG-11 medium 20 - Anabaena cylindrical Nutrient medium 8 - Nonheterocystous Oscillotoria Miami BG7 Medium-A except NH4Cl 90 0.3 Photosynthetic bacteria Rhodobacter sphaeroides Minimal medium 5 - Rhodobacter capsulatus Minimal medium 25 - Rhodobacter palustris Rhodospirillaceae medium 24.9 - Rhodospirillum rubnum CO & H2O 358b - Fermentative bacteria Enterobacter aerogenes Glucose 390 390 - Enterobacter cloacae IIT-BT 08 Sucrose 660 6.0 Clostridium butyricum Glucose 205 - Citrobacter spY19 Glucose - 2.49 Bacillus coagulans Glucose - 2.28

-159- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Clostridium acetobutylicum ATCC 824 Glucose 220 - a ml/mg Chl a.l. b ml/g cell.h

Combined photosynthetic and anaerobic The morphology of thermophilic anaerobic bacterial hydrogen production: hydrogen producing bacteria was rod and Anaerobic bacteria metabolize sugars to enodspore-formation. From DGGE fingerprint, produce hydrogen gas and organic acids, but the GC content of thermophilic anaerobic are incapable of further breaking down the hydrogen producing bacteria was higher than organic acids formed. The combined use of mesophilic anaerobic hydrogen producing photosynthetic and anaerobic bacteria for the bacteria (Chang, 2001). In addition to several conversion of organic acids to hydrogen. known species of the genera Bacteroides, Theoretically, one mole of glucose can be Clostridium, Enterococcus, Escherichia, converted to 12 moles of hydrogen (Fig. 1) Eubacterium and Klebsiella, two strains which through the use of photosynthetic bacteria classified based on phenotypic and phylogenetic capable of capturing light energy in such a considerations in a new genus Dorea as Dorea combined system. From a practical point of longicatena sp. nov.. Experiments with a specific view, organic wastes frequently contain sugar or 16S rRNA directed oligonucleotide indicate that sugar polymers. It is not however easy to obtain D. longicatena sp. nov. is present in all human organic wastes containing organic acids as the volunteers studied so far at average cell counts main components. The combined use of of 1,55 x 109/g of dry weight faces. They form photosynthetic and anaerobic bacteria should with 0,58% of the total cell count a considerable potentially increase the likelihood of their proportion of the total flora. (Taras, 2001). application in photobiological hydrogen Thermophilic bacteria that are able to production. utilize CO under strictly anaerobic conditions,

producing H2 and CO2 are of potential interest in astrobiology. Many volcanic exhalations

contain high levels of CO2, and anaerobic, CO-utilizing, hydrogen-producing thermophiles can be isolated on the Kuril and Kermadec Islands, south of the Kamchatka Peninsula, in terrestrial hydrothermal springs (Bonch- Osmolovskaya et al., 1999). Carboxydothermus. Figure (1): Free energy changes in hydrogen-producing reactions by anaerobic an photosynthetic bacteria hydrogenoformans, the prototype for these strains, was characterized as a Gram positive Ecology of Hydrogen-producing thermophile with the potential for H Bacteria: 2 production (Svetlichny et al., 1991, 1994). Since

-160- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

then such isolates have been obtained from deep organisms, however, the reaction is mediated by sea hydrothermal vents (Sokolova et al., 2001) proteins coordinated in an enzymatic pathway. and hot springs in Yellowstone National Park, The reaction takes place at low (ambient) USA. C. hydrogenoformans was the first strictly temperature and pressure. Thermodynamics of carboxydotrophic strain to be described, with the reaction are very favorable to CO-oxidation an optimal growth temperature of 72°C and H2 synthesis since the equilibrium is (Svetlichny et al., 1991). Later, another species, strongly to the right of this reaction.

Carboxydothermus restrictus was isolated from Stoichiometric amounts of CO2 and H2 are hydrothermal outlets on Raoul Island on the produced during CO-oxidation. The enzyme Kermadec Archipelago (Svetlichny et al., 1994). that binds and oxidizes CO, carbon monoxide: These strains grow optimally at 75°C, using CO acceptor oxidoreductase (carbon monoxide as an energy and carbon source, with H2 and dehydrogenase = CODH) is part of a membrane

CO2 as the only end detectable products. bound enzyme complex (Wakim and Uffen, Chemolithotrophic growth by means of CO 1982). Rubrivivax gelatinosus CBS is a purple oxidation is coupled to hydrogen and carbon non-sulfur bacterium that not only performs dioxide formation according to the following CO–water–gas shift reaction in darkness, equation: converting 100% CO in the atmosphere into

near stoichiometric amounts of H2, it also CO + H2O —> CO2 + H2 ∆G = -20 kJ/mole assimilates CO into new cell mass I, the light The sequence of Carboxydothermus (via CO2 6xation) when CO is the sole source of hydrogenoformans confirmed. The earlier carbon (Maness and Weaver, 1997). Even when observation that the strain was capable of spore an organic substrate is available with CO, R. formation, for it has a set of genes encoding for gelatinosus CBS will utilize both substrates each of the major stages of endospore formation simultaneously, indicating that the CO- (DiRuggiero et al., 2002). oxidation pathway is fully functional even when Hydrogen synthesis via the water–gas a more favorable substrate is included. R. shift reaction of photoheterotrophic gelatinosus CBS exhibits a doubling time of 7 h bacteria: in light when CO serves as the only carbon Certain photoheterotrophic bacteria within source (Maness and Weaver, 2002). The mass the superfamily Rhodospirillaceae can grow in transfer of CO may be enhanced by a high ratio the dark using CO as the sole carbon source to of gas phase to liquid bacterial suspension, and generate ATP with the concomitant release of by stirring the culture vigorously. The

H2 and CO2 (Champine and Uffen, 1987). The hydrogenase from this organism is tolerant to oxidation of CO to CO2 with the release of H2 O2, exhibiting a half-life of 21 h when whole occurs via water–gas shift reaction. In these cells were stirred in full air (Maness et al., 2002).

-161- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

A specific rate of CO oxidation to H2 production industry produces highly concentrated, of 0:8 mmol/min/g of cells, dry weight (cdw) was carbohydrate-rich wastewaters. It can use for measured using a low-density culture (final hydrogen production. Biohydrogen was OD660 ،0:2), stirred at a high rate (250 rpm), produced by a domestic wastewater that had and supplemented with 20% CO in the gas apple, potato processing and confectioners with phase. Because the conversion of CO to H2 is both. Biogas produced consistently contained stoichiometric, this corresponds to a rate of CO 60% hydrogen, with the balance as carbon uptake and conversion of approximately 1:34 g dioxide. (Van Ginkel et al., 2005). CO/h/g cdw, or 48 mmol CO/h/g dcw. An Starch based wastewater has great

OD660 of 2.0 yields 2:0 g R. gelatinosus CBS potentiality for the H2 production (Yu et al., cdw/l. This corresponds to a H2 production rate 2002). The major problem of using industrial of 96 mmol H2/2 g cdw/h or 96 mmol H2/l/h). wastewater is the presence of components in the Advantages and disadvantages of different reaction mixture. Ruminococcus albus has been hydrogen production processes are shown in found suitable for the production of H2 from Table (2). energy crop such as sweet sorghum by utilizing its free sugar, cellulose and hemicelluloses Biological hydrogen production (Ntaikou et al., 2008). H yield is varied from resources: 2 0.47 to 2.52 mol/mol glucose in continuous and Cost of the raw materials play a very batch experiments, respectively. important role for the overall economy of the Microcrystalline cellulose (Wang et al., hydrogen generation process. There are various 2008) and corn stover biomass pretreated with a applications where the process of biological steam-explosion process are found suitable hydrogen production by cyanobacteria can be substrates for the hydrogen production (Datar well utilized. The examples can be included et al., 2007). The H2 yields of 2.84 and 3.0 are from food and chemical industries, which obtained using the mixed sugar present in the employ the process of hydrogenation to produce hydrolysates derived from neutral and acidic derivatives that are used as food additives, steam explosion, respectively. Delignified wood commodities, and fine chemicals (Dutta et al., fiber is found to produce an average yield of 1.6 2005). Biological hydrogen production from the mol H2/mol glucose by Clostridium fermentation of renewable substrates is one thermocellum 27405 in a batch system (Levin et promising alternative although the use of al., 2007). Comparison between the activity of commercially produced food products, such as some organisms applied with different wastes corn and sugar, is not yet economical are shown in Table (3).

(Benemann, 1996). Also hydrogen produced from sweet sorghum by thermophilic bacteria (Claassen et al., 2004). The food processing

-162- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

دول Table 2: Advantages and disadvantages of different hydrogen production processes from biomass Process Advantages Disadvantages Microbial conversion Can be operated at ambient temperature and atmospheric pressure. . Lower rate of hydrogen production and yield. . Can produce H2 directly from water and sunlight Solar conversion . Requires high intensity of light. Direct biophotolysis energy increased by ten folds as compared. . O2 can be dangerous for the system. . o trees, crops. . Lower photochemical efficiency. . Uptake hydrogenase enzymes are to be removed to stop . Cyanobacteria can produce H2 from water. Indirect biophotolysis degradation of H2. . Has the ability to fix N2 from atmosphere. . About 30% O2 present in gas mixture. . A wide spectral light energy can be used by these bacteria. O has an inhibitory effect on nitrogenase. Photo-fermentation 2 . Can use different organic wastes. Light conversion efficiency is very low, only 1–5% . O is a strong inhibitor of hydrogenase. . It can produce H all day long without light. 2 2 . Relatively lower achievable yields of H . . A variety of carbon sources can be used as substrates. 2 . As yields increase H fermentation becomes Dark fermentation . It produces valuable metabolites such as butyric, lactic and acetic 2 thermodynamically unfavorable. acids as by products. . Product gas mixture contains CO which has to be . It is anaerobic process, so there is no O limitation problem. 2 2 separated.

Table 3: Use of different waste materials for the biohydrogen production processes (cited from; Debabrata and Veziroglu 2008) Maximum rate of Maximum H2 yield Name of the wastes Organism used Process H2 production (mol H2/mol substrate) (ml H2/l h) Dairy wastewater Anaerobic mixed consortia UASB - 0.122a Closdridium butyricum and Enterobacter Starch manufacturing wastes Repeated batch culture - 7.2 aerogenes HO-39 and Rhodobacter sp. M-19 Sewage biosolids Mixed culture Batch - 0.7b Rice winery wastewater Sludge from wastewater treatment plant Up-flow reactor - 2.14 Potato processing wastewater Sludge from wastewater treatment plant Batch 6a MSW Mixed culture PBR 99c Food wastes Sewage sludge Batch 112.2d 5.48a Food wastes Mesophilic and thermophilic cultures Batch 1.8 Jackfruit peel Cow dung Anaerobic contact filter 400e Activated sludge and Rhodobacter Olive mill wastewater Two-stage process 11 - sphaeroides O.U. 001 a mmol H2/g COD. b mol H2/g COD. c mlH2/g VS removed.d mlH2/g VSS h. e ml/g VS destroyed -163- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Wastewaters show great potential for a cyanobacterium, Synechococcus PCC7942, economical production of hydrogen because by developing a genetic engineering system producing a product from a waste could reduce for cyanobacteria. These workers also waste treatment and disposal costs (WERF, demonstrated that clostridial hydrogenase 1999). Different waste materials have been protein, when electro-induced into successfully used in different processes for the cyanobacterial cells is active in producing hydrogen generation (Table 3). Some author hydrogen by receiving electrons produced by observed that sewage sludge in combination photosystems. with molasses improves the hydrogen yield of Another strategy being examined is the the process to a great extend (Yokoi et al., 2002). enhancement of hydrogen-producing capabi- lities of photosynthetic bacteria. In nitrogenase- Enhancement of hydrogen-producing capabilities through genetic engineering: mediated hydrogen-producing reactions, a considerable amount of light energy which is Although genetic studies on photosynthetic converted to biochemical energy by the microorganisms have markedly increased in photosystem, is lost through various recent times, relatively few genetic engineering biochemical processes. Control of the studies have focused on altering the photosystem at an appropriate level for characteristics of these microorganisms, nitrogenase activity, would result in reduced particularly with respect to enhancing the energy losses, and thus improved light energy hydrogen-producing capabilities of conversion. To this end, with the objective of photosynthetic bacteria and cyanobacteria. utilizing genetic engineering techniques in Some nitrogen-fixing cyanobacteria are controlling the photosystem level in the potent potential candidates for practical hydrogen hydrogen-producing photosynthetic bacteria production. Hydrogen production by Rhodobacter sphaeroides RV, the puf operon nitrogenase is, however, an energy-consuming encoding photoreaction center and light- process due to hydrolysis of many ATP harvesting proteins was isolated and molecules. On the other hand, hydrogenase- characterized (Nagamine et al., 1996). dependent hydrogen production by cyanobacteria and green algae is "economic" in As most organic substrates undergo that there are no ATP requirements. This combustion with the evolution of energy, the mechanism of hydrogen production is not biocatalyzed oxidation of organic substances by however sustainable under light conditions. oxygen or other oxidizers at two-electrode Water-splitting by hydrogenase is potentially an interfaces provides a means for the conversion ideal hydrogen-producing system. Asada et al., of chemical to electrical energy. Abundant (1986a; 1986b) attempted to overexpress organic raw materials such as methanol, hydrogenase from Clostridium pasteurianum in organic acids or glucose can be used as

-164- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

substrates for the oxidation process, and temperature and corrosive nature of the molecular oxygen or H2O2 can act as the electrolyte makes them unsuitable for use in substrate being reduced. Intermediate mobile and transportation applications. Molten formation of hydrogen as a potential fuel is carbonate fuel cells (MCFC) utilize carbonate -3 possible as well. ions (CO2 ) as the mobile ion, operate at o Escherichia coli, Enterobacter aerogenes, approximately 650 C, and can take H2, CO2,

Clostridium butyricum, Clostridium CO, and/or CH4 as fuel, which means they can acetobutylicum, and Clostridium perfringens are use natural gas, coal gas, or biogas as fuel examples of bacteria used in microbial biofuel sources. Like PAFC, MCFC are used as cells. On the other hand, Alcaligenes eutrophus stationary power sources, generating electricity Escherichia coli, Anacystis nidulans, Proteus in the MW range. Solid oxide fuel cells (SOFC) - vulgaris, Bacillus subtilis, Pseudomonas putida, utilize oxygen radicals (O2 ) as the mobile ion Pseudomonas aeruginosa and Streptococcus and operate between 500oC and 1000oC. Like lactis are examples of bacteria used in biofuel MCFC, SOFC can utilize H2, CO, and/or CH4 as cells upon application of membrane-penetrating fuel, which means they can use methane, coal electron transfer mediators. gas, or biogas as fuel sources. Carbon dioxide is Fuel cells are electrochemical devices that not utilized as a fuel and is discharged as a create an electron flow using charged ions. A waste gas. Like other high-temperature fuel cell variety of different fuel cells systems have been systems (PAFC and MCFC) SOFC systems are developed. They differ in the type of electrolyte used as stationary power sources, generating used, in the operating conditions, in their power electricity from the low kW to the MW range. density range, in their application, and each has PEMFC utilize hydrogen protons (H+) as the its advantages and disadvantages (reviewed by mobile ion, operate in the 50–100oC range,

Larminie and Dicks, 2000). Alkaline fuel cells require pure H2 and are extremely sensitive to (AFC) utilize hydroxyl ions (OH-) as the mobile the presence of CO. Of all the fuel cell systems ion (derived from potassium hydroxide, KOH), that are available, PEMFC systems are operate in the 50 to 200oC range, and are especially suitable for mobile and extremely sensitive to the presence of CO2. transportation applications, and PEMFC Phosphoric acid fuel cells (PAFC) utilize engines have been demonstrated successfully in protons (H+) as the mobile ion and operate at both cars and buses. Small PEMFCs, in the approximately 200oC. PAFC systems were the 1–10 kW range, are also under commercial first fuel cells produced commercially and are development as small stationary power units to used as stationary power sources, generating up provide electricity to homes and small to 200 kW of electricity. Many are in use in the businesses. Because of their imminent USA and in Europe. The high operating commercial applications, PEMFC technologies

-165- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

are under intense research and development. while a bioreactor of approximately 1250 l (1247

The rate of H2 consumption by PEMFCs is l,) would provide sufficient H2 to power a 5:0 usually expressed in kg of H2/s or in mol of H2/s. kW PEMFC. By way of comparison, the current state-of-the-art for distributed, on-site

Biohydrogen: prospects for practical production of hydrogen is via stationary application: electrolyzers which can generate H2 from H2O The analyses of Levin et al., (2004) indicate at a rate of 1000 l/h, or 40:3 mol/h. Equivalent that photosynthesis-based systems do not rates of H2 synthesis could be achieved by a produce H2 at rates that are sufficient to meet bioreactor of approximately 334 l using a dark- the goal of providing enough H2 to power even a fermentation system, or by a bioreactor of 1 kW PEMFC on a continuous basis. This does approximately 420 l containing R. gelatinosus not mean that these systems should be CBS. The CO-water shift reaction of R. abandoned. There may be applications other gelatinosus CBS offersan additional advantage. than our hypothetical objective to which they The CO–water shift reaction also yields equi- may b more suited. Moreover, continued molar amounts of CO2, which R. gelatinosus research will no doubt result in significant CBS can assimilate into new cell biomass using improvements in their respective technologies, organic compounds, such as butyrate or and thus in the rates of H2 production. propionate, as a carbon source and ATP as an Thermophilic and extreme thermophilic energy source (Madigan et al., 1996). Thus, in biohydrogen systems would require bioreactors addition to reformation of CO and H2 synthesis, in the range of approximately 2900–14; 600 l to this process could be incorporated into an provide sufficient H2 to power PEMFCs of 1.5– integrated energy system to sequester CO2 in

5:0 kW, and a bioreactor of approximately 5700 light, after H2 is recovered from the system. l would be required to power the 5:0 kW fuel Bioreactors of 1000–1500 l in the basement of a cell using the in particular appears most home are not unthinkable. Before electrical and promising. A bioreactor of approximately 500 l natural gas heating, it was normal for North

(495 l) would provide enough H2 to power a 2:5 American homes located in northern latitudes to kW PEMFC, while a bioreactor of have, in their basements, tanks of heating oil approximately 1000 l (989 l) would provide that were approximately this size, while dark- sufficient H2 to power a 5:0 kW PEMFC. The fermentation systems and the CO-water were CO–water shift reaction of R. gelatinosus CBS shift. is intriguing as it offers the potential to capture Reaction may have practical applications, and reform CO, and produce H2. A bioreactor there are a number of technical challenges that of approximately 624 l would be required must be considered and overcome before these provide enough H2 to power the 2:5 kW EMFC, systems can be used to produce H2 to power a

-166- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

PEMFC. The most significant of these problems Fermentative bacteria usually produces is whether the systems can be scaled up to hydrogen under acidic conditions (pH 4.5–6.5), volumes large enough to generate the required but photosynthetic bacteria works well at pH flow rate (22:1 mol H2/h for the 5:0 kW fuel above 7 (Younesi et al., 2008). Partial pressure cell). The working volume of the bioreactor of H2 effects the efficiency of the process to a described that produced 121 mmol H2/l/h) was great extends (Mandal et al., 2006). The energy 3l, and used sucrose (at 20 g/l of culture) as the conversion efficiency by photosynthetic bacteria carbon source for bacterial growth (Chang et is inversely proportional to the intensity of light al., 2002). The biogas produced was 25–35% H2 (Nath and Das, 2005). and 65–75% CO2. Further research is required Chemically; addition of oxygen to an H2- to determine if the rate of H2 production will evolving culture, as well as the addition of remain at high levels if these systems are scaled nitrate to cells (which had formed the up to much larger volumes (183 l or more), and dissimilatory nitrate reductase system during if carbon sources other than pure sucrose can be the preceding growth), caused immediate used. The major challenge for the CO–water cessation of hydrogen evolution (Kuhn et al., shift reaction is the problem of mass transfer: 1984). Fertilizer helped gas production because the gas must be available to the bacteria, in of nitrates and phosphates, iron filings, and heat solution, at a sufficient concentration that the in the form of sunlight all, while lye (pH approx. bacteria can absorb and metabolize efficiently. 9) and lemon juice (pH approx. 5) prevented gas This may require radically new technologies production (Dreszer, 2005). Comparing the and bioreactor designs. addition of sodium and potassium ion for thermophilic hydrogen producing bacteria, the Effect of physico-chemical parameters: hydrogen producing bacteria were more The effect of several physicochemical sensitivity with potassium ion. parameters on the photo-biological processes Substrate concentration affects the has already been reported (Das and Veziroglu, fermentative H2-production processes to a great 2001). Physically; Temperature, pH and extent. Malate and glutamate play important medium composition play very important roles roles in this fermentation process. The initial for the H2 production. The lower temperature acetic acid concentration present in the spent and lower pH value would make the activity of medium of dark fermentation process has a hydrogen producing bacteria decrease (Chang, profound effect on hydrogen production.

2001). The fermentative H2 reduction carried Acetate concentration up to 55 mM is found out at different temperatures: 20, 30, 35 and non-toxic to the photofermentation of hydrogen

55°C indicated that higher temperatures tend to (Nath and Das, 2005). H2 production is found to improve the H2 yield. (Yu et al., 2002). be increased through redirection of metabolic

-167- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

pathways by blocking formation of alcohol and power fuel cells of sufficient size to do practical some organic acids in E. cloacae (Kumar et al., work. 2001). Future prospects: Hydrogen production from cellulose by -The future of biological hydrogen production microflora is performed by a consortium of depends on research advances, and several species of microorganisms. (Ueno et al., improvement in efficiency through genetically 2001). engineering microorganisms and/or the development of bioreactors. Concluding remarks: -Progress on biological hydrogen production Biohydrogen technologies are, still in their processes economically is still not attractive as infancy. Thus, further research and compared to the conventional H2-production development aimed at increasing rates of processes. synthesis and final yields of H are essential. 2 -Hydrogen production depends also on There are many technical challenges, from the economic considerations (the cost of fossil production of sufficient quantities of hydrogen fuels), social acceptance, and the development to its storage, transmission, and distribution. of hydrogen energy systems. So that the One of the major limitations to the practical following points require immediate attention: application of biohydrogen systems is that  Improvement of H2 yield of the processes scientists who study biohydrogen systems do not using cheaper raw materials, talk to engineers who develop hydrogen fuel cell  Development of mixed microbial consortia or technologies (and vice versa). Thus, the rates of metagenomic approaches may be used to hydrogen reduced by biological systems are develop efficient microbial strains for the unknown to fuel cell engineers and the amounts better utilization of industrial wastewater, of H2 required for practical applications, such which has different carbon content. as fuel cells, are unknown to biohydrogen

 Optimization of bioreactor designs, rapid researchers. The rates of hydrogen produced by removal and purification of gases, and genetic the various biohydrogen systems are expressed modification of enzyme pathways that in different units, making it difficult to assess compete with hydrogen producing enzyme and compare the rates and mounts of hydrogen systems offer exciting prospects for synthesized by different biohydrogen biohydrogen systems. technologies. Existing technologies offer  In two-stage processes, the major bottleneck potential for practical application, but if lies on the photo-fermentation process. biohydrogen systems are to become Improvement of these processes surely will commercially competitive they must be able to improve overall hydrogen yield as well as synthesize H2 at rates that are sufficient to economy of the process.

-168- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

REFERENCES: Claassen P.A.M., de Vrije T. and Budde M.A.W. (2004): Biological hydrogen Asada Y. and Kawamura S.J. (1986a); production from sweet sorghum by Screening for cyanobacteria that evolve thermophilic bacteria. 2nd. World molecular hydrogen under dark and Conference on Biomass for Energy, anaerobic conditions. J. Ferment. Tech., Industry and Climate Protection. Rome, 64: 553-556. Italy. Asada Y. and Kawamura S. J. (1986b): Aerobic Das D., and Veziroglu T.N. (2001): Hydrogen Hydrogen Accumulation by a Nitrogen- production by biological processes: a Fixing Cyanobacterium, Anabaena sp. survey of literature. Int. J. Hydrogen App. Environ. Microbiol., 51:1063-1066. Energy. 26:13–28. Benemann J. (1996): Hydrogen biotechnology: Datar R., Huang J, Maness PC, Mahagheghi A, progress and prospects. Nature Czernik S and Chounet E. (2007): Biotechnol 14:1101-1114 Hydrogen production from the Bolton J.R. (1996): Solar photoproduction of fermentation of corn stover biomass hydrogen. Sol Energy. 57:37–50. pretreated with a steam explosion Bonch-Osmolovskaya E.A, Miroshnichenko process. Int. J. Hydrogen Energy. M.L., Slobodkin A.I., Sokolova T.G., 32:932–9. Karpov G.A., Kostrikina N.A., Zavarzina Debabrata D. and Veziroglu T.N. (2008): D.G., Prokofieva M.I., Rusanov I.I. and Advances in biological hydrogen Pimenov N.V. (1999): Distribution and production processes. Int. J. Hydrogen biodiversity of anaerobic lithotrophic Energy. 33:6046–6057. prokaryotes in terrestrial hydrotherms of Di Ruggiero J., Nandakumar R., Eisen J.A., Kamchatka. Mikrobiologia. 68: 680-686. Schwartz M., Thomas R., Davila J., Champine J.E, and Uffen R,L. (1987): Memb- Ofman L. and Robb F.T. (2002): rane topography of anaerobic carbon Genomic and physiological studies on monoxide oxidation in Rhodocyclus extremophiles: model systems for gelatinosus. J. Bacteriol. 169: 4784–9. exobiology. Workshop «Astrobiology in Chang J.S., Lee K.S. and Lin P.J. (2002): Russia». Petersburg, Russia. Biohydrogen Production with fixed-bed Dreszer T.B. (2005): The Art of Brewing bioreactors. Int. J. Hydrogen Energy. Hydrogen: Improving Gas Yield of 27:1167–74. Hydrogen-Producing Bacteria. California Chang S.M. (2001): Research On Membrane State Science Fair. Project No. 25032. Bioreactor Applied For Thermophilic Dunn S. (2002): Hydrogen futures: toward a Anaerobic Hydrogen Production (MSc sustainable energy system. Int. J. thesis, English abstract). Hydrogen Energy. 27:235–64.

-169- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Dutta D., Debojyoti D., Surabhi C. and Levin D.B., Islam R., Cicek N. and Sparkling R. Bhattacharya S.K. (2005): Hydrogen (2007): Sparkling hydrogen production production by Cyanobacteria, Review. by Clostridium thermocellum 27405 from Microbial Cell Factories. 4:36 cellulosic biomass substrate. Int. J. Fascetti E. and Todini O. (1995): Rhodobacter Hydrogen Energy. 31:1496–503 sphaeroides RV cultivation and hydrogen Levin D.B., Pitt L. and Love M. (2004): production in a one- and two-stage Biohydrogen production: prospects and chemostat. Appl Microbiol Biotechnol. limitations to practical application. Int. J. 22:300–5. Hydrogen Energy. 29:173–85. Fedorov A. S., Tsygankov A. A., Rao K. K. Liu J., Bukutin V.E. and Tsygankov A.A.

and Hall D.O. (1998): Hydrogen (2006): Light energy conversion into H2 photoproduction by Rhodobacter by Anabaena variables mutant PK84 sphaeroides immobilised on polyurethane dense culture exposed in nitrogen foam. Biotechnol Lett. 20:1007–9. limitations. Int. J. Hydrogen Energy. Francou N. and Vignais P.M. (1984): Hydrogen 31:1591–6. production by Rhodopseudomonas Lopes P.F.A., Troshina O. and Lindblad P. capsulata cells entrapped in carrageenan (2002): A brief look at three decades of beads. Biotechnol Lett. 6:639–44. research on cyanobacterial hydrogen Jeong T.Y., Cheol C.G., Yeom S.H. and Suk evolution. Internat. J.Hydrogen Energy. S.C. (2008): Comparison of hydrogen 27:1209-1215. production by four representative Madigan M.T., Martinko J.M. and Parker J. hydrogen-producing bacteria. J. Indus. (1996): Biology of microorganisms. Eng. Chem. 14:333–337. Upper Saddle River, NJ: Prentice-Hall. Kuhn M., Steinbochel A. and Schlege H.G. Mandal B., Nath K. and Das. D. (2006): (1984): Hydrogen evolution by strictly Improvement of biohydrogen production

aerobic hydrogen bacteria under under decreased partial pressure of H2 by anaerobic conditions. J. Bacteriol. 159(2): Enterobacter cloacae. Biotechnol. Lett. 633–639. 28:831–5. Kumar N., Ghosh A. and Das D. (2001): Maness P.C. and Weaver P.F. (1997): A Redirection of biochemical pathways for potential bioremediation role for

the enhancement of H2 production by photosynthetic bacteria. In: Sikdal SK, Enterobacter cloacae. Biotechnol. Lett. Irvine RL, editors. Bioremediation: 23:537–41. principlesand practice, Vol. II. Larminie J. and Dicks A. (2000): Fuel cell Technomic Publishing Co, Inc., systems explained. New York: Wiley. Lancaster PA.

-170- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Maness P.C. and Weaver P.F. (2002): Hydrogen Pinto F.A.L., Troshina O. and Lindblad P. production from a carbon-monoxide (2002): A brief look at three decades of oxidation pathway in Rubrivivax research on cyanobacterial hydrogen gelatinosus. Int J. Hydrogen Energy. evolution. Int. J. Hydrogen Energy. 27: 27:1407–11. 1209–15. Maness P.C., Smolinski S., Dillon A.C., Heben Prince R.C. and Kheshgi H.S. (2005): The M.J. and Weaver P.F. (2002): photobiological production of hydrogen: Characterization of the oxygen tolerance potential efficiency and effectiveness as a of a hydrogenase linked to a carbon renewable fuel. Crit. Rev. Microbiol. monoxide oxidation pathway in 31:19–31. Rubrivivax gelatinosus. Appl. Environ. Rey E.F., Heiniger E.K. and Harwood C.S. Microbiol. 68: 2633–6. (2007): Redirection of Metabolism for Momirlan M. and Veziroglu T.N. (2002): Biological Hydrogen Production. Appl. Current status of hydrogen energy. Environ. Microbiol. 73(5):1665–1671 Renew Sustain Energy Rev. 6:141–79. Simpson F.B. and Burris R.H. (1984): A Nagamine, Y. et al., (1996): J. Marine Biotech. nitrogen pressure of 50 atmospheres does 4:34-37. not prevent evolution of hydrogen by http://www.fao.org/docrep/w7241e/w724 nitrogenase. Science. 224:1095–1097. 1e0g.htm Sokolova T.G., Gonzalez J.M., Kostrikina N.A., Nandi R. and Sengupta S. (1998): Microbial Chernyh N.A., Tourova T.P., Kato C., production of hydrogen: an overview. Bonch-Osmolovskaya E.A. and Robb Crit. Rev. Microbiol. 24:61–84. F.T. (2001): Carboxydobrachium Nath K. and Das D. (2003): Hydrogen from pacificum gen. nov., sp. nov., a novel biomass. Curr. Sci. 85:265–71. anaerobic, thermophilic, CO-utilizing Nath K. and Das D. (2005): Hydrogen marine bacterium from Okinawa production by Rhodobacter sphaeroides Trough. Int. J. Syst. Appl. Bact. 51:141- strain O.U. 001 using spent media of 149. Enterobacter cloacae strain DM11. Appl. Svetlichny V.A., Sokolova T. G., Kostrikina Microbiol. Biotechnol. 68:533–41. N.A. and Lysenko A.M. (1994): Ntaikou I., Gavala H.N., Komaros M. and Carboxydothermus restrictus sp. nov. a Lyberatos G. (2008): Hydrogen new thermophilic anaerobic carboxydotr- production from sugars and sweet ophic bacterium. Mikrobilogiya, 63: 523- sorghum biomass using Ruminococcus 528. albus. Int. J. Hydrogen. Energy. 33: Svetlichny V.A., Sokolova T.G., Kostrikina 1154–63. N.A., and Zavarzin G.A. (1991): Anaerobic extremely thermophilic

-171- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

carboxydotrophic bacteria in microcrystalline cellulose using coculture hydrotherms of Kuril Islands. Microbial –Clostridium acetobutylicum X9 and Ecology. 21: 1-10. Ethanoigenens harbinense B48. Int. J. Taras D. (2001): Intestinal hydrogen Hydrogen. Energy. 33:912–7. metabolism: In vitro experiments and Water Environment Research Foundation establishment of agnotobiotic rat model- (WERF) (1999): Research needs to Datum der Promotion: (English optimize wastewater resource utilization. Abstract). Project 98- CTS-1. Tsygankov A. A., Hirata Y., Miyake M., Asada Winkler M., Hemsehemeier A., Gotor C., Melis Y. and Miyake J. (1994): Photobioreactor A. and Happe T. (2002): [Fe]- with photosynthetic bacteria immobilized hydrogenase in green algae: photo- on porous glass for hydrogen photop- fermentation and hydrogen evolution roduction. J. Ferment. Bioeng. 77:575–8. under sulfur deprivation. Int. J. Ueno Y., Haruta S., Ishii M. and Igarashi Y. Hydrogen Energy. 27:1431–9. (2001): Microbial community in Yokoi H., Maki R., Hirose J. and Hayashi S. anaerobic hydrogen-producing micro- (2002): Microbial production of hydrogen flora enriched from sludge compost. Appl from starch-manufactering wastes. Microbiol Biotechnol 57, 555–562. Biomass Bioenergy. 22:389–95. Van Ginkel S.W., Oha S.E. and Logana B.E. Younesi H., Najafpour G., Ismail K.S.K., (2005): Biohydrogen gas production from Mohamed A.R. and Kamaruddin A.H. food processing and domestic (2008): Biohydrogen productionin a wastewaters. Internat. J. Hydrogen continuous stirred tank bioreactor from Energy 30:1535 – 1542. synthesis gas by anaerobic photosynthetic Vincenzini M., Materassi R., Sili C. and bacterium: Rhodospirillum rubnum. Florenzano G. (1986): Hydrogen produ- Bioresour. Technol. 99:2612–9. ction by immobilized cells III. Prolonged Yu H., Zhu Z., Hu W. and Zhang H. (2002):

and stable H2 photoevolution by Hydrogen production rice winery Rhodopseudomonas palaustris in light- wastewater in an upflow anaerobic dark-cycles. Int. J. Hydrogen Energy. reactor by using mixed anaerobic 11:623. cultures. Int. J. Hydrogen Energy. 27: Wakim B.T. and Uffen R.L. (1982): Membrane 1359-65. association of the carbon monoxide Zurrer H. and Bachofen R. (1979): Hydrogen oxidation system in Rhodopseudomonas production by the photosynthetic gelatinosa. J. Bacteriol. 153:571–3. bacterium, Rhodospirillum rubrum. Appl. Wang A., Ren N., Shi Y. and Lee D.J. (2008): Environ. Microbiol. 37:789–93. Bioaugmented hydrogen production from

-172- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

دور الطحالب الدقيقة و البكتريا في إنتاج الهيدروجيـن كإحدى مصادر الطاقة المتجددة

أحمد محمد على عبد المنعم، فرج محمد على شعيب

قسم النبات – كلية العلوم – جامعة عمر المختار – البيضاء - ليبيا

غاز الهيدروجين يعد حاليا واحدا من أهم مصادر الطاقة البديلة للبترول. ولقد أتضح أنه ليس فقط من الوقود الغير ملوث، بل أنه يحد من انبعاثات ثانى أكسيد الكربون ومنتجا لغاز الهيدروجين كواحد من مصادر الطاقة المتجددة. إن التناقص المستمر في الوقود الحفري يلزمه إيجاد مصادر جديدة وغير تقليدية للطاقة عوضا عن تلك التي تتمثل في الوقود الحفري. ولقد ثبت أن الكائنات النباتية الدقيقة لها القدرة على إنتاج غاز الهيدروجين من خالل عملية البناء الضوئي معتمدة على ضوء الشمس كمصدر طبيعي ودائم للطاقة. وتشمل تلك الكائنات الطحالب الخضراء والخضراء المزرقة )السيانوبكتريا(، من خالل عملية االختزال الضوئي المباشر والغير مباشر. باإلضافة إلى أن تلك الطحالب الخضراء المزرقة تنتج الهيدروجين خالل تحلل المركبات العضوية )التحلل الضوئي(. ويعتبر إنتاج الهيدروجين من الطحالب الخضراء من الوسائل االقتصادية والدائمة، وكذلك استخدام المياه واعادة استخدام ثاى أكسيد الكربون من المصادر المستدامة للطاقات النظيفة. كما أن معدالت إنتاج الهيدروجين باستخدام بكتريا التحلل الضوئي من الخاليا الثابتة أكثر منه في العالقة. وتعد الطحالب الخضراء المزرقة من أفضل الكائنات الدقيقة إلنتاج الهيدروجين، كما انه باال مكان تطبيقاتها على المستوى التجاري. وهي أفضل اقتصاديا مقارنة بالطرق التقليدية مثل الكيميائية والكهروضوئية. هذا البحث المرجعي يوضح األسس البيولوجية إلنتاج الهيدروجين من الطحالب الدقيقة والبكتريا والقاء الضوء على بعض التوقعات المرئية لتطبيق هذه السبل. ويتوقف استخدامها على الجهد المعرفي والتكنولوجيات ومدى التقدم والتطور المستقبلي الواجب قبيل اعتمادها كمصدر أساسي للطاقة.

-173- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

INVESTIGATION OF RESIDUAL PESTICIDES IN SOME VEGETABLE PLANTS IN SAUDI ARABIA Abdullah Saati Assistant Professor of Environmental Health & Toxicology Consaltant of Occupational Hygiene Dept. of Community Medicine and Pilgrims Healthcare, Faculty of Medicine, Umm Al-Qura University

ABSTRACT: Pesticides are used for pest management and vector control in agricultural areas, but many farming communities are not adequately informed about the hazards associated with the chemicals. As a result, farmers use pesticides without full understanding of their impact on human health and the environment. Human contacts with pesticides, whether in the field, during pesticide application, weeding, pruning, harvesting, re-entry to collect fire wood, vegetables or in the houses for killing mosquitoes, cockroaches, fleas and flies. Storing pesticides may lead into acute and/or chronic exposures, with adverse health consequences. Although the inhalation, dermal and oral routes of exposure are the most common, pesticide residues in food and water may add to indirect exposures common in the general population. This study aimed to develop and evaluate a rapid automated screening method for determining pesticides residues in seven green vegetables (rocket, mint, parsley, leek, dill, lettuce and spinach) collected from different sources in Saudi Arabia markets. Thirty nine samples from green vegetables were used in this study. Six types of green vegetables were taken from the open markets (16 samples) and seven types were taken from supermarket (21 samples). These samples were screened for pesticides residues by using Turbo Matrix Headspace Trap system in conjunction with a Perkin Elmer Clarus soo GC/MS. The classes of pesticides which are detected in this study are herbicides, rodenticides and fungicides. The obtained results revealed that the Dinitrotoluidine [1- hydroxy-4-(p-toluidine) anthraquinone] was detected in rocket and parsley, which are presents in supermarket but not found in mint, dill, lettuce and spinach. The Oxadiazolinon (N, O- dimethyloxos tephene) herbicide was found only in rocket purchased from open market. It was found that the dill obtained from the open market was the only plant which contains all kinds of rodenticides like cyano-phenolic (3-cyano-6-triflorom ethylphenantherine), sodium flouroacetate (3-methyl-1-phenyl-2- azafluorenone), hydrogen cyanide (1-cyano-6-trifloromethylphenantherine) and arsenical (arsine,1,2-phenylenebis dimethyl). Hydrogen cyanides were detected in dill only. Identified types of pesticides have shown more occurrences in open market samples than the supermarket samples. The contamination level of pesticide residues considered a possible public health problem. Regular monitoring of a greater number of samples for pesticide residues, especially the imported is needed.

-13- INTRODUCTION: (Glotfelty and Schomburg, 1989). Drift can account for a loss of 2 to 25% of the chemical I has been observed that a diet containing being applied, which can spread over a distance fresh fruit and vegetables far outweigh potential of a few yards to several hundred miles. As risks from eating very low residues of pesticides much as 80–90% of an applied pesticide can be in crops (Brown, 2004). Increasing evidence volatilised within a few days of application (Dietary Guidelines, 2005) shows that eating (Majewski, 1995). fruit and vegetables regularly reduces the risk Commonly used pesticides include of many cancers, high blood pressure, heart herbicides insecticides rodenticide and disease, diabetes, stroke, and other chronic fungicides. All pesticides are toxic by their diseases. nature and they cause health hazards to human If the credits of pesticides include enhanced and animals. There are concerns that pesticides economic potential in terms of increased used to control pests on food crops are production of food and fibre, and amelioration dangerous to people who consume those foods. of vector-borne diseases, then their debits have These concerns are one of the reasons for the resulted in serious health implications to man organic food movement. Many food crops and his environment. There is now including fruits and vegetables contain pesticide overwhelming evidence that some of these residues after being washed or peeled. chemicals do pose a potential risk to humans Chemicals that are no longer used are resistant and other life forms and unwanted side effects to breakdown for long periods may remain in to the environment (Forget, 1993; Igbedioh, 1991; Jeyaratnam, 1985). No segment of the soil and water and so in food (NAS, 1987). population is completely protected against Selective herbicides kill specific targets exposure to pesticides and the potentially while leaving the desired crop relatively serious health effects, though a disproportionate unharmed. Some of these act by interfering with burden, is shouldered by the people of growth of the weed and are often synthetic developing countries and by high risk groups in "imitation" of plant hormones. Herbicides used each country (WHO, 1990). The world-wide to clear waste ground, industrial sites, railways deaths and chronic diseases due to pesticide and railway embankments are non-selective and poisoning number about 1 million per year kill all plant material with which they come into (Environews Forum, 1999). contact. Herbicides represent about 70% of all Pesticide sprays can directly hit non-target agricultural pesicides used in USA (Kellogg et vegetation, or can drift or volatilize from the al., 1999). They include dinitrotoluidine, treated area and contaminate air, soil, and non- oxadiazolinon, quinoline derivatives, cyano- target plants. Some pesticide drift occurs during phenoxypentenoic acid, nitrophenolic and every application, even from ground equipment phenolic compounds. Fungicides are used both in agriculture and to fight fungal infections in

-14- animals. Fungicide residues have been found in samples were shown in Table 1. Each sample food for human consumption, mostly from post- washed with 5 ml distilled water. Sample harvest treatment (Brooks and Roberts, 1999). preparation was done according to the methods Oral exposure is the most severe hazardous of EPA (2007). route for pesticide poisoning in human occur by 2-Head Space GC/MS: accident of ingestion of contaminated food, Headspace sampling provides easy through accidental splashing of pesticides in the extraction of volatile compounds from liquid face and mouth, or by rubbing the face with and solid samples, while eliminating the time contaminated hands or gloves. The degree of consuming and error-producing steps required toxicity and hazard from ingestion of pesticides in other GC sample-preparation techniques is related to the toxicity of the materials, their (March, 2000; Matz et al., 2002 and Erasmus et solubility and the affected portion of the al., 2005). gastrointestinal tract (Jiguo et al., 2007). Inhalation of pesticide dusts, vapors, mists 3-Calibration and Standardization: and gases may represent a significant The MS was set up for electron impact (EI) occupational hazard. Dust hazards may involve ionization at the 8260B-specified 70eV and a full the loading, mixing, and application of Auto Tune generated was performed and saved. insecticides in powder or granular form. 25% of The Auto Tune generated parameters are the inhaled pesticide is exhaled, 25% is shown. After Auto Tune, the multiplier was deposited on the upper respiratory passages and decreased to 2350 V and the emission current subsequently swallowed and 25% is deposited decreased to 75 μA because the headspace trap on the lung (Anwar, 1997). provided so much sensitivity that the MS

sensitivity could be reduced to prolong filament MATERIALS AND METHODS: life, increase stability and extend linearity 1- Samples: concentrations. Using the standard Auto Tune Thirty nine samples from green vegetables parameters without modification had produce were used in this study. Six types of green acceptable results (EPA, 2007). vegetables were taken from the open markets (18 samples) and seven types were taken from supermarket (21 samples). Types and sources of

Table 1: Types and sources of samples Type of samples Open market Super market Total Rocket 3 3 6 Parsley 3 3 6 Mint 3 3 6

-15- Leek 3 3 6 Dill 3 3 6 Lettuce 3 3 6 Spinach 0 3 3 Total 18 21 39

-16- Table 2: Gas Chromatography Settings Gas Chromatograph PerkinElmer Clarus 500 GC Headspace Connector Universal connectror Oven Program Initial Temperature 40˚C Holt time 1 2 min Ramp 1 10˚C/min to 100˚C Hold time 2 0 min Ramp 2 30 ˚C/min to 240˚C Hold time 3 5 min Equilibration time 0.5 min Vacuum compensation On Headspace control On Column Elite volatiles-30 m X 250 μm X 1.4 μ film Carrier gas Helium

Table 3: Mass Spectrometry Settings Mass Spectrometer PerkinElmer Clarus 500 MS Mass range 35-300 u Solvent Delay Time 0 min Scan Time 0.1 sec Interscan Delay Time 0.1 sec Transfer Line Temperature 200 ˚C Source Temperature 200 ˚C Multiplier 350 V Trap Emission 75 μm Threshold 0 Software Turbo Mass 5 with reporting

Table 4: Headspace Settings Sample introduction PerkinElmer Turbo Matrix HS-110 Trap Needle Temperature 90˚C Transfer Line Temperature 120˚C Oven Temperature 80˚C Trap Low Temperature 40˚C Trap High Temperature 280˚C Dry Purge (Helium) 5 min Trap Hold Time 6 min Desorb Time 0.5 min Thermostatting_ Time 10 min Pressurization Time 1 min Decay Time 2 min Outlet Split 20 ml/min Column Pressure 25 psi Vial Pressure 35 psi Desorb Pressure 10 psi Transfer Line Deactivated Fused silica 20 m x 320 μm

RESULTS: not found in mint, dill, lettuce and spinach. The Oxadiazolinon (N, O-dimethyloxos tephene) 1-Herbicides: herbicide was found only in rocket purchased The obtained results revealed that from open market. The other herbicide the Dinitrotoluidine [1-hydroxy-4-(p-toluidine) Quinoline Deriveative (2-hydroxynaphthy1- anthraquinone] are positive in rocket and 2-quinoline), Cyanophenolic (3-cyano-6- parsley, which are presents in supermarket but trifloromethylphenan therine), Nitrophenolic

-17- (2-methoxy-1-nitrophenazine-5-oxide and 3- all kinds of rodenticides like cyano-phenolic acridinol-9-phenyl) and phenolic [phosphine, (3-cyano-6-triflorom ethylphenantherine), methyl bis 4-N-N-dimethyl amino) phenyl, sodium flouroacetate (3-methyl-1-phenyl- 1-methyl-3-phenyl-4-azafluorenone, oxazole, 2- azafluorenone), hydrogen cyanide (1-cyano-6- 2-1-naphthalenyl-5-phenyl, 2,2-bis 4-methoxy- trifloromethylphenantherine) and arsenical phenyl-2-ethoxy-ethane and silane, dimethyl-2- (arsine,1,2-phenylenebis dimethyl) (Table 6). propenyl tetradecyloxy) were not found in any leaves except the dill which are present in open 3-Fungicides: market (Table 5). Hydrogen cyanides were detected in dill

only (Table 7).

2-Rodenticides: It was found that the dill obtained from the open market was the only plant which contains

Table 5: Types of herbicides detected in green leaves Pesticide scientific Types of green leaves Organic name name R P M L D L S Dinitrotoluidine 1-hydroxy-4-(p-toluidine) anthraquinone +ve +ve -ve -ve -ve -ve -ve Oxadiazolinon N,O-dimethyloxos tephene +ve -ve -ve -ve -ve -ve -ve Quinoline derivatives 2-hydroxynaphthy1-2-quinoline -ve -ve -ve -ve +ve -ve -ve Cyano-phenilic 3-cyano-6-trifloromethylphenantherine -ve -ve -ve -ve +ve -ve -ve 2-methoxy-1-nitrophenazine-5-oxide -ve -ve -ve -ve +ve -ve -ve Nitrophenolic 3-acridinol-9-phenyl -ve -ve -ve -ve +ve -ve -ve Phosphine [methyl bis 4-N-N-dimethyl amino) -ve -ve -ve -ve +ve -ve -ve phenyl] 1-methyl-3-phenyl-4-azafluorenone -ve -ve -ve -ve +ve -ve -ve Phenolic oxazole [2-1-naphthalenyl-5-phenyl] -ve -ve -ve -ve +ve -ve -ve 2,2-bis 4-methoxy-phenyl-2-ethoxy-ethane -ve -ve -ve -ve +ve -ve -ve silane [dimethyl-2-propenyl tetradecyloxy)] -ve -ve -ve -ve +ve -ve -ve

Table 6: Types of rodenticides detected in green leaves Pesticide Scientific Types of green leaves Organic name name R P M L D L S Cyano-phenilic 3-cyano-6-trifloromethylphenantherine -ve -ve -ve -ve +ve -ve -ve Sodium flouroacetate 3-methyl-1-phenyl-2- azafluorenone -ve -ve -ve -ve +ve -ve -ve Hydrogen cyanide 1-cyano-6-trifloromethylphenantherine -ve -ve -ve -ve +ve -ve -ve Arsenical arsine,1,2-phenylenebis dimethyl -ve -ve -ve -ve +ve -ve -ve

Table 7: Types of fungicides detected in green leaves Pesticide Scientific Types of green leaves Organic name name R P M L D L S Hydrogen cyanide 1-cyano-6-trifloromethylphenantherine -ve -ve -ve -ve +ve -ve -ve Arsenical arsine,1,2-phenylenebis dimethyl -ve -ve -ve -ve +ve -ve -ve Rocket (R), Parsley (P), Mint (M), Leek (L), Dill (D), Lettuce (L), Spinach (S).

-18- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

DISCUSSION: pesticides remaining on food product samples. Maximum residual level is maximum quantity Food quality and safety are receiving of pesticide that may still be present on the crop increasing attention in the food sector nationally at point of safe that is legally permitted in, or on and internationally. Emerging private sector crops or a food commodity, allowed by national regulations are increasingly perceived as governments which are set to determine legal market entrance barriers within this trading limit that are not an indicator of risk to development all over the world (Morland et al., health. It ensures the safety of food for 2002). The growing use of pesticides accounts consumers and regulates international trade. for a significant portion of the increase in food Multi-residue methods are required to monitore productivity over the past several decades. pesticide residues. The analytical performance Pesticides clearly have assisted in controlling was demonstrated by the analysis of extracts pests and maintaining the availability of low from lettuce, orange, apple, cabbage, grape and cost and high quality food. Risk of dietary wheat flour, spiked at three concentration levels exposure to pesticides residues also has been of ranging from 0.01 to 0.10 mg/Kg pesticide or its public concern while solid scientific evidence metabolite (Maurice and Andre, 2007). Gas has not been completely established. Pesticides chromatography (GC) with selective detection in food were considered to be the most coupled to mass spectrometry (GC-MS) are the hazardous to human health (EPA, 1986). analytical methods which more frequently Crops (vegetables and fruits) are an applied to the analysis of pesticide residues in important part of a healthy diet as they are a foods, fruit, vegetables and cereals (Ana et al., significant source of vitamins and minerals. 2004). Quality of these crops can be affected by pest as In this study identification of the type of part of nature profile. However, these pests can pesticides present in vegetables in Saudi Arabia be controlled by (Dayan et al., 1999). Pesticides is addressed. GC/MS technique was used in this constitute a very important group of highly study for this reason. In addition, one of the toxic compounds, therefore usage of pesticides most useful techniques was used in this study is in agriculture practice and public should be headspace trap. Headspace trap can omit the controlled. The levels of pesticide residues in step for sample preparation which makes the foodstuffs are harmful to human based on a techniques used in this investigation is more long range exposure (intake). Previous studies faster and reliable. Headspace technique can found that there was an association between increase the sensitivity of the analysis as there is pesticide in crops and cancer (NAS, 1987). no addition of extraction solvents that can dilute Pesticides residues on crops are monitored the target compound concentration in the with reference to Maximum Residue limits and sample. are based on analysis of quantity of a given

-19- Results of the present study revealed that 2003) it was revealed that farmers were not dinitrotolidine is positive in supermarket rocket receiving agricultural extension service hence and parsley, while oxadiazolinon is positive only have attempted various means especially in in open market rocket. The other herbicides pesticides use when dealing with pest problems (quinoline derivatives, nitrophenolic and but were constrained by the lack of appropriate phenolic) are positive in open market dill. These knowledge. However, pesticide usage in the results are consistent with previous findings in study area seems to be highly influenced by which herbicides are found in crops (Conde et manufacturers and pesticides vendors who were al., 2006 and Zheng-Min et al., 2006). carrying out their business right in the farming Fungicides and rodenticides were not communities and very interested in achieving detected in all types of investigated green leaves large sales of their pesticides. This is a typical of vegetables except dill obtained from open situation in many developing countries where market which showed positive results for the choice of pesticides to be used by farmers is fungicides and rodenticides. Phosphine influenced by the suppliers (Snoo et al., 1997; poisoning increases levels of superoxide Epstein and Bassein, 2003). In African dismutase and malondialdehyde in non- countries, many government extension survivors, while catalase was inhibited (Chugh programs encourage the use of pesticides (Abate et al., 1996). Oxidation of phosphine can lead to et al., 2000), but do not consider their effects in formation of reactive phosphorylating species the environment and health risks. As a result (Lam et al., 1991), thus suggesting that effects and coupled with lack of basic knowledge of on cholinesterase may be possible (Garry and et pesticides, farmers’ decisions on what pesticides al., 2001). Studies of grain fumigant applicators and how to use do not have a bearing on health (Potter et al., 1993) and in vitro studies of or safety of the environment. Epstein and human red blood cells (Potter et al., 1991) have Bassein (2003) observed that farmers used more shown that significant phosphine-induced pesticides because they based the applications inhibition of red blood cell cholinesterase occurs on calendar spray pesticides program without at concentrations of phosphine exceeding 10 necessarily giving much priority to health and μg/ml. environmental considerations. The use of pesticide was observed to be The high dependence on pesticides by high, with over 40 different formulations, vegetable farmers is an indication that they are probably because farmers assume that the only not aware of other pest management strategies solution to pest problems is to spray more that are effective, inexpensive and yet friendly frequently and using different types of to the environment. Pest management strategies pesticides (Dinham, 2003). In previous studies including intercropping (Legutowska et al., conducted in some of the study areas (Ngowi, 2002) and tillage type and crop rotation

-20- (Hummel et al., 2002) have been shown to Pendimethalin [N-(1-ethylpropyl)-3,4- dimethyl significantly reduce insect pests. There is a need -2,6-dinitrobenzenamine], in the form-ulation of to bring to the attention of those farmers Prowl (a commercial herbicide), was applied to existing alternative pest management strategies various crops. Analysis of pendimethalin and its that are cost effective and environmentally metabolite [4(1-ethylpropyl)amino-2-methyl- friendly. 3,5-dinitrobenzyl alcohol] was accomplished by The obtained results are in agreement utilizing liquid-liquid partitioning, gel with those obtained by Hall et al. (2004). Who permeation chromatography (GPC) for nuts found that methoxyfenozide [3-methoxy-2- and mint, solid-phase extraction (SPE) cleanup, methylbenzoic acid 2-(3,5-dimethylbenzoyl)-2- and gas chromatography (GC) with a nitrogen-- (1,1-dimethylethyl) hydrazide; RH-2485], in the phosphorus detector (NPD). Method validation formulation of INTREPID, was applied to recoveries for fruits, nuts, vegetables, grass, and various crops. Analysis of methoxyfenozide was mint are given for both compounds. accomplished by utilizing liquid-liquid Pendimethalin average recoveries ranged from extraction and partitioning, followed by liquid 71% to 126% over two levels of fortification. chromatography-tandem mass spectrometry Pendimethalin metabolite average recoveries (LC-MS/MS). Method validations for fruits, ranged from 69% to 123% over two levels of vegetables, and mint are reported. fortification. The quantitation limit for all crops Methoxyfenozide mean recoveries ranged from except mint was 0.050 ppm. The quantitation 72 to 129% over three levels of fortification. The limit for mint and mint oil was 0.10 ppm. overall average of mean recoveries is 97+10%. Residues greater than the limit of quantitation The limit of quantitation for fruits, artichoke, were found for pendimethalin in apple pomace, cucumber, squash, and refined sugar was 0.010 fresh and dry fig, grass screenings, mint oil, ppm, with a detection limit of 0.005 ppm. For all almond hulls, green onion, and tomato pomace other crops, the limit of quantitation was 0.050 (wet and dry). Residues greater than the limit of ppm, with a detection limit of 0.025 ppm. No quantitation were found for pendimethalin residues were found greater than the limit of metabolite in grass screenings, grass straw, and quantitation in control samples. Residues above almond hulls. All other crop analysis for the limit of quantitation were found in all pendimethalin and its metabolite were below the matrices except refined sugar. Foliage (bean, limit of quantitation (Engebretson et al., 2001). beet, pea, and radish) had greater residue levels Dissipation rates of boscalid [2-chloro of methoxyfenozide residue than their -N-(4'-chlorobiphenyl-2-yl) nicotinamide], corresponding roots or pods. Other crop pyraclostrobin [methyl 2-[1-(4-chlorophenyl) matrices contained<1.0 ppm of methoxyfenozide pyrazol-3-yloxymethyl]-N-methoxycarbanilate], except artichoke, which had a mean of 1.1 ppm. lufenuron [(RS)-1-[2,5-dichloro-4-(1,1,2,3,3,3-

-21- hexafluoropropoxy) phenyl]-3-(2,6- and urban pesticide uses contribute to the difluorobenzoyl)urea] and lambda-cyhalothrin problem. Pesticide contamination poses [(R)-cyano (3-phenoxyphenyl) methyl (1S,3S)- significant risks to the environment and non- rel-3-[(1Z)-2-chloro-3,3,3-trifluoro-1-propenyl]- target organisms ranging from beneficial soil 2,2-dimethylcyclopropanecarboxylate] in green microorganisms, to insects, plants, fish, and beans and spring onions under Egyptian field birds. Contrary to common misconceptions, conditions were studied. Field trials were even herbicides can cause harm to the carried out in 2008 in a Blue Nile farm, located environment. In fact, weed killers can be at 70 kilometer (km) from Cairo (Egypt). The especially problematic because they are used in pesticides were sprayed at the recommended relatively large volumes. The best way to reduce rate and samples were collected at pre- pesticide contamination (and the harm it causes) determined intervals. After treatment (T(0)) the in our environment is for all of us to do our part pesticide residues in green beans were 7 times to use safer, non-chemical pest control methods. lower than in spring onions. This is due to a different structure of vegetable plant in the two CONCLUSION: crops. In spring onions, half-life (t(1/2)) of Identified types of pesticides have shown pyraclostrobin and lufenuron was 3.1 days and more occurrence in open market samples than 9.8 days, respectively. At day 14th (T(14)) after the supermarket samples. treatment boscalid residues were below the The contamination level of pesticide Maximum Residue Limit (MRL) (0.34 versus residues could be considered a possible public 0.5 mg/kg), pyraclostrobin and lambda- health problem. The results also emphasize the cyhalothrin residues were not detectable (ND), need for regular monitoring of a greater while lufenuron residues were above the MRL number of samples for pesticide residues, (0.06 versus 0.02 mg/kg). In green beans, at especially the imported ones. T(0), levels of boscalid, lufenuron and lambda- GC/MS is an accurate, fast, precise and cyhalothrin were below the MRL (0.28 versus 2 easy-to-maintain technique which is used to mg/kg; ND versus 0.02 mg/kg; 0.06 versus 0.2 identify drugs, pesticides, environmental mg/kg, respectively) while, after 7 days analysis, unknown samples of food, beverages treatment (T(7)) pyraclostrobin residues were and perfumes. Headspace is a unique technique above the MRL (0.03 versus 0.02 mg/kg). in which sample preparation is easy and However, after 14 days the residue level could convenient. There is no need to empty the lines go below the MRL (0.02 mg/kg), as observed in to remove residual contamination and there is spring onions (Hanafi et al., 2010). Pesticide no risk of sample foam contamination of purge- residues are found in soil and air, and on and –trap device. surface and ground water across the countries,

-22- Public awareness about pesticides can be compounds released by hydrothermal increased by more appropriate education treatments of olive tree pruning. Food programs based on pesticide control should be Chemistry, 114 (3): 806-812. initiated at national level. Dayan, F.E.; Romagni, J.G.; Tellez, M.R.; Rimandi, A.M. and Duke, S.O. (1999): REFERENCES: Pesticides. Outlook 5. p. 185. Abate T, van Huis A, Ampofo JKO. (2000): Pest Dietary guidelines for Americans (2005): U.S. management strategies in traditional Department of Health and Human agriculture: An African perspective. Services U.S. Department of Agriculture. Annu. Rev. Entomol., 45:631–659. Dinham B. (2003): Growing vegetables in Ana, A; Susana, L.; Amedeo, R.; Fernandez developing countries for local urban Alba; Mariano, C.; Juan, C. and Luis, P. populations and export markets: proble- (2004): One-year routine application of a ms confronting small-scale producers. new method based on liquid Pest Manag. Sci., 59(5): 575–582. chromatography-tandem mass spectro- Engebretson, J.; Hall, G.; Hengel, M. and metry to the analysis of 16 multiclass Shibamoto, T. (2001): Analysis of pesticides in vegetable samples. J. of pendimethalin residues in fruit, nuts, Chromatography A, 1045 (1-2): 125-135. vegetables, grass, and mint by gas Anwar, W.A. (1997): Biomarkers of human chromatography. J. Agric Food Chem. 49 exposure to pesticides. Environmental (5): 2198-206. Health Perspective Supplements, 105 (4): Environews Forum (1999): Killer environment. 801-806. Environ Health Perspect. 107:A62.

Brooks, G.T. and Roberts, T.R. (1999): EPA (Environmental Protection Agency) (1986): Pesticides Chemistry and Bioscience Office of Pesticides and Toxic Substances. edited. Published by the Royal Society of Guidance for the registration of pesticide Chemistry. products containing glyphosate as the Brown Ian (2004): UK Pesticides Residue active ingredient. Washington DC.

Committee Report. EPA (Environmental Protection Agency) (2007): Chugh, S.N.; Arora, V.; Sharma, A. and Chugh Measuring environmental volatile K. (1996): Free radical scavengers & organic compounds by U.S. EPA method lipid peroxidation in acute aluminium 8260B with headspace trap GC/MS. phosphide poisoning. Indian J Med Res. Perkin Elmer life and Analytical 104: 190–193. Sciences. Conde, E.; Cara, C.; Moure, A.; Ruiz, E., Epstein, L. and Bassein, S. (2003): Patterns of Castro, E. and Domingeuz, H. (2006): pesticide use in California and the Antioxidant activity of the phenolic implications for strategies for reduction

-23- of pesticides [Review] Annu. Rev. cyhalothrin residue levels on green beans Phytopathol. 41: 351–375 and spring onions in Egypt. J. Environ Erasmus, C.; Tara, B. and Brindle, D. (2005): Sci. Health B. 45 (6): 493-500. Headspace gas chromatography-mass Hummel RL, Walgenbach JF, Hoyt GD, spectry: a fast approach to the Kennedy GG. (2002): Effects of identification and determination of production system on vegetable 2-alkyl-3-methoxypyrazine pheromones. arthropods and their natural enemies. The Royal Society of Chemistry, 130: Agric. Ecosyst. Environ. 93(1-3):165-176. 152-155. Igbedioh, S.O. (1991): Effects of agricultural Forget G. (1993): Balancing the need for pesticides on humans, animals and higher pesticides with the risk to human health. plants in developing countries. Arch. In: Forget G, Goodman T, de Villiers A, Environ. Health.;46:218 editors. Impact of Pesticide Use on Jeyaratnam J. Health problems of pesticide Health in Developing Countries. IDRC, usage in the third world. B M J. (1985); Ottawa: 2. 42: 505. Garry VF, Lyubimov AV. Phosphine (2001): In: Jiguo, W.; Li, L.; Tiangang, L.; Yuk, S.; Chan, Krieger RI, editor. Handbook of G. and Chongyu, L. (2007): Effects of Pesticide Toxicology. 2. Vol. 2. San Diego: organophosphorus pesticides and their Academic Press; pp. 1861–1866. ozonation byproducts on gap junctional Glotfelty, J. and Schomburg, J. (1989): intercellular communication in rat liver Volatilization of pesticides from soil in cell line. Food and Chemistry Toxicology, Reactions and Movements of organic 45 (10): 2057-2063. chemicals in soil. In: Sawhney BL, Brown Kellogg, R.L.; Nehring, R.; Grube, A.; plotkin, K, editors. Madison, WI: Soil Science S.; Goss, D.W. and Wallace, S. (1999): Society of America Special Pub. Trends in the potential for environmental Hall, G. L.; Engebretson, J.; Hengel, M. J. and risk from pesticide loss from farm fields. Shibamoto, T. (2004): Analysis of US Department of Agriculture, Natural methoxyfenozide residues in fruits, Resources Conservation Service. vegetables, and mint by liquid Lam, W.W.; Toia, R.F. and Casida, J.E. (1991): chromatography-tandem mass spectro- Oxidatively initiated phosphorylation metry (LC-MS/MS). J Agric Food Chem. reactions of phosphine. J. Agric Food 52(4):672-6. Chem., 39:2274-78. Hanafi, A.; Garau, V.L.; Caboni. P.; Sarais, G. Legutowska H, Kucharczyk H, Surowiec J. and Cabras, P. (2010): Minor crops for (2002): Control of thrips infestation on export: a case study of boscalid, leek by intercropping with clover, carrot pyraclostrobin, lufenuron and lambda- or bean. In: Paroussi G, Voyiatzis D,

-24- Paroussis E, editors. Proceedings of the Delaney paradox. Washington DC, Second Balkan Symposium on Vegetables Nationa. Academy Press. and Potatoes (579) 3001 Leuven 1, Ngowi, A.V.F. (2003): A study of farmers’ Belgium: International Society Horticul- knowledge, attitude and experience in the tural Science; pp. 571–574. use of pesticides in coffee farming. Afr Majewski, M. (1995): Pesticides in the Newslett on Occup Health and Safety., atmosphere: distribution, trends, and 13:62. governing factors. In: Capel P, (editor). Snoo, G.R.; Jong, F.M.W.; de van der Poll, R.J.; Volume one, Pesticides in the Hydrologic Jansen, S.E.; van der Veen L.J, and System. Ann Arbor Press Inc; p. 118. Schuemie, M.P. (1997): Variation of March, R. (2000): Quadruple ion trap mass pesticides use among farmers in Drenthe: spectrometry: A view at the turn of the A starting point for environmental century. International J. of Mass protection. Med. Fac. Landbouww. Univ, Spectrometry, 1-3: 285-312. Gent. 62/2a:199–212. Matz, L.; Asbury, G. and Hill, H. (2002): Two- Potter, W.T.; Garry, V.F.; Kelly, J.T.; Tarone, dimensional separation with electrospray R.; Griffith, J. and Nelson, R. L. (1993): ionization ambient pressure high- Radiometric assay of red cell and plasma resolution ion mobility spectrometry/ cholinesterase in pesticide appliers from quadrupole mass spectrometry. Rapid Minnesota. Toxicol Appl Pharmacol., Communications in Mass Spectrometry, 119: 150–155. 16 (7): 670-675. Potter, W.T.; Rong, S.; Griffith, J.; White, J. Maurice, H. and Andre, de. K. (2007): and Garry, V.F. (1991): Phosphine- Comprehensive multi-residue method for mediated Heinz body formation and the target analysis of pesticides in crops hemoglobin oxidation in human using liquid chromatography-tendem erythrocytes. Toxicol Lett., 57:37–45. mass spectrometry, J. of Chomatography WHO Geneva (World Health Organization A, 1154 (1-2): 3-25. (1990): Public Health Impact of Morland, K.; Steve, W.; Ana, D.R. and Charles, Pesticides Used in Agriculture. P. 88. P. (2002): Neighborhood characteristic Zheng-Min, Y.; Wei, W.; Ai-Liang, H.; Qing- associated with the location of food stores Fu, Y. and Long, L. (2006): Determina- and food service places, American J. of tion of herbicide ZJ0273 residue in Preventive Medicine, 22 (1): 23-29. rapeseed by radioisotopic tracing NAS (National Academy of Sciences)(1987): method. Food Chemistry, 114 (1): 300- Regulating pesticides in food: the 305.

-25- فحص متبقيات المبيدات في بعض نباتات الخضر في المملكة العربية السعودية عبد هللا ساعاتي

أستاذ صحة البيئة والسموم المساعد قسم طب المجتمع والرعاية الصحية للحجيج – كلية الطب – جامعة أم القرى - المملكة العربية السعودية

مبيدات اآلفات يمكن أن تلوث التربة والهواء والماء وغيرها من النباتات كونها تستخدم على نطاق واسع لمكافحةة اآلفةات والنواقل في المنةاطق الرراعيةة ولكةن ال توجةد معلومةات كافيةة حةول المخةاطر المرتبطةة بهةذ المةواد الكيميائيةة. ونتيجةة لةذل يسةةتخدم المرارعةةون المبيةةدات دون فهةةم كامةةل لتأثيرهةةا علةةى صةةحة اننسةةان والبيئةةة. ويةةؤدى االتصةةال المبا ةةر بةةين اننسةةان والمبيةةدات الح ةةرية سةةواء فةةي المةةرار خةة ل اسةةتخدام المبيةةدات والتع ةةيب والتقلةةيم والحصةةاد واعةاد دخةةول لجمةةع الحطةةب والخضروات أو قتل البعوض والصراصير والبراغيث والذباب في المنارل وكذا تخةرين المبيةدات إلةى التعةرض الحةاد أو المةرمن لهذ المواد السامة مما ينجم عنه عواقب ضار بالصحة. وعلى الرغم مةن أن التعةرض للمبيةدات يمكةن أن يحةدث عةن طريةق االستن اق والجلد والفم والتى تعتبةر أكثةر طةرق التعةرض ةيوعا إال أن التعةرض لمتبقيةات المبيةدات فةي األغذيةة والميةا يةتم بطريقة غير مبا ر في عموم السكان. تهدف هذ الدراسة إلى تطوير وتقيةيم طريقةة لتحديةد بقايةا المبيةدات فةي سةبع أنةوا مةن الخضةروات )الجرجيةر والنعنةا والبقدونس والكراث وال بت والسبانخ والخس( تم جمعها من مصادر مختلفة في أسواق المملكة العربية السعودية. اسةةةتخدمت فةةةي هةةةذ الدراسةةةة تسةةةع وث ثةةةون عينةةةة مةةةن الخضةةةروات. منهةةةا سةةةتة أنةةةوا مةةةن الخضةةةروات مةةةن األسةةةواق المفتوحة )81 عينة( وسبعة أنوا من السوبر ماركت )18 عينة(. تم فحص هذ العينات لمتبقيات المبيدات باستخدام جهةار A Perkin Elmer Clarus 600 MS/GC . وأصناف المبيدات التي تم الك ف عنها في هذ الدراسة هي مبيدات األع ةاب ومبيدات القوارض ومبيدات الفطريات. وجةةةد أن المبيةةةد الع ةةةبى Dinitrotoluidine ]8 - 4- هيدروكسةةةي )ف طولويةةةدين( غليكوسةةةيد[ يوجةةةد فةةةي الجرجيةةةر والبقدونس التي تعرض في مح ت السوبر ماركت ولكن ال يوجد في النعنا وال بت والخس والسبانخ. كمةةا تةةم العثةةور علةةى المبيةةد الع ةةبي )Oxadiazolinon )tephene dimethyloxos - O ، N فقةةط فةةي جرجيةةر التةةي تةةم ةةراؤها مةةن السةةوق المفتوحةةة. أمةةا بخصةةوص مبيةةدات القةةوارض فقةةد وجةةد أن ال ةةبت الةةذ يةةتم الحصةةول عليةةه مةةن السةةوق المفتوحةةة. هةةو النبةةات الوحيةةد الةةذ يحتةةو علةةى جميةةع أنةةوا مبيةةدات القةةوارض مثةةل الفينةةول cyano -3( - cyano - -6 flouroacetate )ethylphenantherine triflorom الصةةةةةةوديوم )3 - 8- الميثيةةةةةةل فينيةةةةةةل - azafluorenone - 1( وسةةةةةةةةةةيانيد الهيةةةةةةةةةةدروجين )trifloromethylphenantherine -6 - cyano - 8( ومركبةةةةةةةةةةات الةةةةةةةةةةررنيخ )أرسةةةةةةةةةةين 1 8 phenylenebis - ثنائي ميثيل(. كما تم الك ف عن سيانيد الهيدروجين في ال بت فقط من المبيدات الفطرية. ولقد أظهرت نتائج الدراسة وجود أنوا محدد من المبيدات في عينات السةوق المفتوحةة أكثةرمن عينةات السةوبر ماركةت. ويمكن النظر في مستوى التلوث من مخلفات المبيدات مما تسببه من م اكل صحية عامة. والتأكيد أيضا على ضرور الرصةد المنتظم لعدد أكبر من العينات لمتبقيات المبيدات وخصوصا المستورد منها.

-26-

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

SYNTHESIS, SPECTROSCOPIC AND POTENTIOMETRIC STUDIES ON A SILVER(I) COMPLEX DERIVED FROM A NEW BIS(THIOSEMICARBAZONE) Mahmoud Hassan Moustafa*, Ahmed Abd-Elnaeem and Osama Ahmed Abbas Department of Chemistry, Faculty of Science, Al-Azhar University, Assiut Branch, Assiut, Egypt *Author to whom corresponence should be addressed: E- mail: [email protected]

ABSTRACT: A new reagent, (2E,2'Z)-2,2'-(3-((E)-(2-hydroxyphenyl)diazenyl)pentane-2,4- diylidene)bis (hydrazinecarbothioamide) (OPTS) was synthesized and studied. Acid-base, spectrophotometric properties of OPTS were studied in 50% ethanol-water mixture solutions at pH 3–11. Ionization constants of the reagent OPTS have been determined by a spectrophotometric and potentiometric

titrations: pK1=9.60±0.02; pK2=10.45±0.05 in its computational and graphical versions at an ionic

strength 0.1 M NaClO4. The stability constants of the Ag-OPTS are logK1=4.55±0.04 and

logK2=3.92±0.05. The reaction of Ag(I) with OPTS gives a mononuclear complex in 50% ethanol -

water mixture solution with λmax=462 nm at pH=8.1. The effects of foreign ions and masking agents on the determination of Ag(I) with the new reagent are studied. The mono complex obeys the Beer`s law in the Ag(I) concentration range 1.1–17 mg/25ml. Molar absorption coefficients were determined. In this paper, we report the synthesis of the novel Ag(I) complexes with OPTS. The complexes were characterized by UV-Vis and IR spectroscopy, elemental analysis, molar conductivity and thermal decomposition.

INTRODUCTION: Domagk[8]. This paper describes the synthesis of Thiosemicarbazones and their metal one of the newer bis(thiosemicarbazone) complexes have become the subjects of reagents (2E,2'Z)-2,2'-(3-((E)-(2-hydroxy important studies because of their wide ranging phenyl)diazenyl)pentane-2,4-diylidene) bis biological activities (antibacterial [1], (hydrazinecarbothioamide) (OPTS). The acid- antimalarial[2,3], antiviral[4], analytical base equilibria of the ligand (OPTS) have been applications and interesting chemical and investigated. We also describe here the synthesis structural properties[5–7]. Antitumor activity of and some physico-chemical and structural thiosemicarbazone were reported first by characteristics of its first prepared complex

-27- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

with Ag(I). The bis-Schiff base ligand, H2L, buffer solutions, at pH 7.0 followed by a pH 4.0 synthesized by condensing o-hydroxyphenyl azo at 25ºC by coupling the titration cell with a acetylacetone with thiosemicarbazide, by thermostatic bath set at this temperature. refluxing in a stoichiometry of 1:2. The first The electronic spectra of solutions of the Ag(I) complexes with thiosemicarbazide-based ligand OPTS and its silver complex were derivatives have raised considerable interest recorded on a Perkin-Elmer (Lambda 35) due to their analytical properties. On the basis computerized spectrophotometer equipped with of elemental analysis, absorption spectra and 1 cm matched quartz cells. IR-spectra were potentiometry, the metal to ligand stoichiometry recorded using a Jasko 480 Fourier transform of 1:1 has been proposed for these complexes. infrared spectrometer (FT-IR) analysis in the The proposed structure for the complex has region 400-4000 cm-1 using the potassium the support by IR and UV-visible spectra. bromide disk technique. Elemental analysis were carried out by the unit of micro analyses Experimental : on a Perkin–Elmer 240 C instrument. Molar 1-Reagents: conductivities of freshly-prepared 1x10–3M All chemicals were Analar chemically pure solutions were measured on a Jenway 4320 grade. O-aminophenol, acetylacetone and conductivity meter. NMR spectra were recorded thiosemicarbazide used to prepare the ligand in DMSO on Varian EM-360 L, 60 MZ NMR OPTS were purchased from Sigma-Aldrich spectrometer and JEOL FX 90 Q Fourier Chemicals Co., USA and were used as received. transform NMR. Mass spectra were performed The metal salt silver nitrate (AgNO3) was by Shimadzu- GCMS-OP 1000 Ex using direct purchased from Merck (Germany) and was inlet system. Thermogravimetric analysis TGA used without purification. Perchloric acid were carried out with Schimadzu DTG-60 H (HClO4), sodium perchlorate (NaClO4), Sodium Simultaneous DTA-TG apparatus at heating hydroxide (NaOH), potassium hydrogen rate of 10oC/min in nitrogen atmosphere. phthalate (KHC8H4O4), Borax 3-Preparations of the ligand: (Na2B4O7.10H2O), DMF and pure ethanol of

Analar products were obtained from Merck o-Hydroxyphenyl azo acetyl acetone (o- (Germany). HPAA) was synthesized by the known

procedure[9], adopting a reported earlier 2-Physical measurements: procedure[10]. The reaction of two molecules of pH measurements were carried out using a diamines and ketone gives a compound Corning 215 pH meter with a combined glass containing two imino groups ―CH=N― in each electrode. The glass electrode was calibrated unit. Preparation of the ligand (2E,2'Z)-2,2'-(3- before each titration with two Merck standard ((E)-(2-hydroxyphenyl)diazenyl)pentane-2,4-

-28- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

diylidene) bis(hydrazinecarbothioamide) d6 solvent of the (a) Schiff base, H2L, ligand (b)

(OPTS) is depicted in Scheme 1. A solution of o- Schiff base, H2L, ligand after addition of D2O hydroxyphenyl azo acetyl acetone (0.02 M) in 20 (*)-NH, NH2 and OH bands which disappeared ml of ethanol was continuously stirred with an after the addition of D2O solvent. Where (a) ethanolic solution of thiosemicarbazide (0.04 NMR for ligand, D2O NMR for ligand,) b(notes M). The reaction mixture was heated under that the disappearance for (OH) and (NH) in reflux for 6 h on a water bath at 60ºC. The Fig. 2(b). resulting solution mixture was kept for one day. After this time, the formed yellowish powder of 4-Preparation of silver (I) complexes: the bis(thiosemicarbazone) ligand was collected. A general procedure was followed: An The solid formed was filtered and washed with equimolar amounts of a warm aqueous (20 mL) cold EtOH and Et2O and dried in vacuo. H2L: solution of corresponding metal salt (0.01 mol.) Anal. Found: C, 42.82; H, 4.30; N, 29.97; S, was added to a warm ethanolic solution (20 mL)

17.47. C13H18N8OS2 calculate; H, 4.31; N, 29.98; of the ligand OPTS (0.01 mol). The mixture was S, 17.48%; mass spectrum: m/z 365.77 (Fig. 1), heated under reflux with stirring at an adjusted yields 76% m.p 290ºC. 1H NMR spectra of the pH value of 8.1 for 6–7 h. On cooling a black Schiff base ligand (Fig. 2) were performed in complex precipitated out, which was filtered DMSO and the chemical shifts in ppm were washed with cold ethanol and dried in vacuo recorded as follows: δ=2.4 ppm (s, 3H, CH3– over P4O10 (yield 71%). AgL: Anal. Found: C,

C=N), δ=11.35 ppm (s, 1H, (s, 1H, OH), δ=10.45 32.75; H, 3.58; N, 23.64; S, 6.77, C13H17ON8AgS2 ppm (s, 2H, NH–C=S), δ=7.66 ppm (s, 2H, NH2– calculate: C, 32.99; H, 3.59; N, 23.68; S, 6.76.

C=S), δ=7.27 , 7.25, 7.01 ,6.95 ppm (m, 4H, Ar– H). Fig. 2. 1H NMR spectra (δ, ppm) in DMSO-

OH OH

S H N N NH2 N N N N O H2N S + 2 N NH2 + 2H2O H N O HN S

NH2 Scheme 1: Preparation of the OPTS (H2L) ligand

-29- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Fig. 1: Mass spectrum of the bis(thiosemicarbazone)(OPTS)

Fig. 2: 1H NMR spectra of the bis(thiosemicarbazone)(OPTS)

Fig. 3: FT-IR spectrum of the bis(thiosemicarbazone)(OPTS)

-30- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

RESULTS AND DISCUSSION: thiosemicarbazides anion in which the sulfur [12,13] 1- pH metric studies: atom is anionic . The dianionic form results from deprotonation of the two mercapto groups. The acid-base properties of the bis-Schiff base The potentiometric titration curve for OPTS ligand (2E,2'Z)-2,2'-(3-((E)-(2-hydroxyphenyl) (Fig. 4) in the form H L shows two inflections at diazenyl)pentane-2,4-diylidene) bis 2 m=1 followed by a moderate inflection at m=2 (hydrazinecarbothioamide) OPTS (H2L) in 50% (m=moles of base added per mole of ligand) ethanol-water mixture solution indicated that corresponding the stepwise dissociation the predominant form of the ligand is the constants of the ligand. The titration curve for a neutral species, which undergoes stepwise system containing 1:1 molar ratio of Ag(I) and ionization on increasing the pH of solution. The OPTS exhibits one inflections at m=1 (m= moles protonation equilibria of ligand (H2L) can be of base added per mole of metal ion) indicating represented by equations (1) and (2): the formation of mono-binary complex. - + H2L HL + H ……….. (1)

HL- L-- + H+ ………… (2)

n The A, and pL values were calculated at different pH values using the known equations[11]. The proton dissociation constants were determined by plotting A against the pH values giving the proton ligand formation curve.

The ionization constants of the ligand (logK1 and or logK2) are calculated at A values of 0.5 and 1.5. The values of dissociation constants of

OPTS were found by potentiometry: Fig. 4. Potentiometric titration curves of: A) pK1=9.60±0.02 and pK2=10.45±0.05 in NaClO4 deprotonated ligand OPTS, and B) Ag(I)- OPTS as supporting electrolyte [I=0.1M]. The reagent binary systems. The abscissa represents the moles of alkali added per mole of ligand OPTS is considered as H2L ligand and the following species are involved in the acid-base - - - forms H2L, HL , L (Scheme 2). The central NH The formation and stability of binary group can deprotonate rapidly and delocalise complex with OPTS in 1:1 molar ratio were with the terminal C=S groups, forming an studied by potentiometric pH titration in 50% enamic form in the conjugate anionic ligand. As (v/v) ethanol-water medium, ionic strength a consequence there are several resonance I=0.1M (NaClO4), temp.=25°C. The metal- structures for the N-substituted ligand stability constants were obtained from

-31- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

the curves drawn between n and pL using half in which the metal to ligand ratio was 1:2 and -5 method. The and pL values were calculated the concentration of each metal ion was 8X10 at different points in the pH range of study. M. From the formation curves the stepwise Potentiometric determination of stoichiometry stability constants of binary chelates were of the binary complexes proved that the ligand determined and are listed in Table (1). The (OPTS) form 1:l (M:L) binary complex. The complexed species found in this work for OPTS stability constant for the binary Ag-OPTS and the studied metal ions can be represented complexes were calculated from titration curves by the following Eqs. 3.

M []ML M + L ML , k ML = , ...... (3) [ML ].[ ]

Table 1: The ionization constants of the ligand OPTS and the formation constants of their Ag(I) complexes Compound H+ Ag(I) complex M ML ML pK1 pK2 log k log k log ML ML ML2 2 OPTS 10.45 9.60 4.55 3.92 8.47

βa is the overall stability constant.

The predicted species for the ligand OPTS in the pH range studied are depicted in Scheme 2.

- -- Scheme 2: structural representations of H2L, HL and L

-32- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

2-Electronic spectra: (Fig. 6) in the range 300–400 nm and is assigned to a combination of intraligand and LMCT The spectra of the neutral forms of the absorptions[21]. It can be seen in Fig. 6 that the ligand OPTS were recorded over the available absorption maxima at 462 nm of silver mono region in 50% (v/v) ethanol-water medium, complex exhibit bathochromic shifts relative to from 270 to 800 nm, as well in the presence of the absorption maximum of the reagent. The NaClO [I=0.1 M] as supporting electrolyte. The 4 solution spectra were recorded in equimolar absorption spectra of 2.5X10-4M solution of the solutions and in solutions containing an excess reagent (OPTS) were recorded as A=F(λ) for of the reagent or the metal ions. The absorption various pH's in the presence of 50% ethanol. spectra reflect the formation of one complex The spectra showed an absorption band with species and the existence of chelate equilibrium the pH range 5–9. The strong broad band in the studying pH range. The absorption observed between (300-700 nm). The spectra of spectra for equimolar solutions and those the ligand are characterized by a band at 340 containing an excess of AgI are analogous and nm (Fig. 5). The absorbance versus pH graphs exhibit an absorption band at 462 nm. The were interpreted according to the relationship absorption bands are observed in the pH range reported in literature[14]. The dissociation 4–9.9. The absorption bands characterizing the constants of OPTS were also determined AgI complex species in the pH studied range are spectrophotometrically beside the above study shown in Fig. 6. The absorbance versus pH in water-ethanol and I=0.1 M NaClO . The pK 4 graphs for Ag-OPTS were interpreted using values were calculated from the variation of the relations derived by Sommer et al.[22,23]. The absorbance with pH according to Phillips and absorbance vs. pH graphs were analyzed Merrit[15] and Sommer[16]. The data obtained for graphically as described previously[24] and one each method to estimate of the acidity constants complexation equilibria was established in of OPTS and the complex formation constants solutions of the complex Ag-OPTS species were calculateded using the SUPERQUAD within the pH range studied. The acidity program[17]. The program has been used to constants and stability constants of the complex calculate acidity constants in systems previously were determined and are a most the same as studied[18-20]. The final results for pK values are calculated using potentiometry. The titrations the average of six pairs of independent were carried out at four OPTS/AgI ratios. The titrations. Molar absorption coefficient of the ligand/metal ratio was varried from 4:1 to 1:1. spectra of the complexes were also recorded The absorption spectrum (Fig. 6) shows the under similar conditions (2880 l mol-1 cm-1). The maximum absorbance of the reaction of an prominent strong bands of the reagent OPTS aqueous Ag(I) solution with an ethanolic with the highest absorbance gives molar solution of OPTS at pH 8.1. The absorbance absorption coefficient equal to 14800 l mol-1 cm-1

-33- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

versus pH graph for solution (Eq. 4) were the binary complexes under investigation determined under the same conditions as used limited the optimal reaction conditions for in the experiments for thosw containing Ag- synthesis of the solid complex. The formation OPTS complex indicating the presence of one constant for the following equilibria in binary system only of complex equilibria at wavelength systems containing Ag-OPTS: 462 nm. The effect of pH absorption spectra of

+ + Ag +(OPTS -H2) [Ag(OPTS-H)]+ H ……..………. (4) were calculated considering the relevant data or the acid dissociation constants and the cumulative binary constants.

Fig. 5: Absorption spectra of Ligand (OPTS) pH: 1(3), 2(4), 3(5), 4(6), 5(7), 6(8), 7(8. 5), 8(9) .

A- Stoichiometry of the complexes: complex. The composition of the complexes was s method of continuous variation[25,26] also confirmed by applying the mole ratioۥJob [27] was applied to establish the composition of the method . complex Ag–OPTS. In solutions with Co=CM + -3 -1 B-Calibaration graph and reproducibility: CL= 3.6X10 mol L at pH=8.1, the maximum Under the optimum conditions a linear of Job plot corresponds to a component ratio of calibration graph for Ag-OPTS complex was 1:1 (metal to ligand). The plot of absorbance obtained up to a concentration of 22.62 μg/25 ml versus mole fraction of Ag+ shows a maximum silver with a molar absorptivity of 2600 l mol-1 at 0.5, suggesting the formation of 1:1 (M:L)

-34- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

cm-1 at 462 nm. A Ringbom plot showed that the and 3202 cm-1 which are attributed to optimum concentration range for the asymmetric and symmetric stretching vibration determination of Ag(I) was 1.35 - 19.98 μg/25 of the NH2 functionality. The weak band at 2977 [28] -1 ml. Sandell’s sensitivity index of the reaction cm is assigned to vibrations of υ(C-H). Another was found to be 6.96X10-3 ng cm-1. The relative important band occurs at 1606 cm-1 in the standard deviation (RSD) was 0.72% for nine spectrum of the ligand attributed to υ(C=N) determinations, each having a silver stretching mode which is red shifted by 20 cm-1 concentration 9.75 μg ml-1. The reproducibility in the spectrum of metal complex. This indicates of the method was checked by testing two series the involvement of the N-atom of the of solutions having silver concentrations of 3.84 azomethine in coordination. The thioamide [N- and 8.87 μg ml-1. HCS] group exhibits a band at 1189 cm−1 in the spectrum of the ligand which disappeared in 3- Molar conductivity: complex spectrum due to its coordination through the sulfur atom, in the thione form, of Molar conductivity of the freshly prepared the thioamide groups. The bands at 1486, 1418, Ag-OPTS complex in DMF solution 1322 and 830 cm−1have contribution from [υ corresponds to 1:1 molar ratio was 4.97 ohm (C–H) + υ ], [υ + υ + υ [υ + υ ] cm-1. The data reveal that the complex is none- (N–H) (C-S) (C–N) (C–H)], (C–N) (C- S) and [υ ], respectively[29]. The IR spectra of electrolyte type. (C=S) the ligand shows an absorption bands at 830 4-IR spectra: and 790 cm-1 due to υ(C=S) stretching vibrations. The disappearance of the two bands An evidence for the formation of -1 at 1189 and 790 cm and remaining the band at thiosemicarbazone and the metal complex was 830 cm−1 in complex spectrum indicate that the obtained from the IR spectra. The character of ligand is coordinated with Ag(I) in its thione the IR spectra is in accordance with the form in the solid state. This result proved that established structure of tetradentate diprotic the absence of the participation of NH group in Schiff-base ligand OPTS which can be found in the complexation. The band observed at 458 the thione or thiol forms (Fig. 3) and its Ag- cm-1 is attributed to Ag-N bonding. Thus, the OPTS complex (Fig. 8). Their prominent feature infrared spectra reveal that OPTS is the presence of υ stretching vibration (OH) bis(thiosemicarbazone) behave as dibasic, frequency at 3435 cm-1, this mean that the quadridentate ligand coordinating through phenolic group not coordinated. This band may thione sulphur and azomethine nitrogen. A be overlapped with the bands corresponding to summary of the analytical, FT-IR spectral data the stretching vibrations of H O. The IR 2 appear on Table 2. spectrum of OPTS, shows two bands at 3125

-35-

-4 Fig. 6: Absorption spectra of Ag-OPTS complex: C L=C M=2. 5× 10 M, I=0.10 M (NaClO4), pH 1(5), 2(5.5), 3(7.1), 4(7.5), 5(8.1), 6(8.5)

Fig. 7: Absorption spectra of Ag-(OPTS) binary complex: CL=CM= -3 2.5×10 M, I=0.1 M (NaClO4), λmax=462 nm

-36- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Fig. 8: FT-IR spectrum of complex Ag-OPTS

Table 2: Infrared spectral data for OPTS ligands and its Ag(I) complex (cm-1)

Compound Υ (OH) υ (NH2) υ (C=N) υ )N=N) υ (C=S) υ (N-C=N) υ (C–S) υ (M-N) s3202 1189 OPTS 3435 1608 1486 1418 1067 - as3120 790 Ag-OPTS 3445 - 1628 1506 750 1419 1088 458

5-Thermal analysis: second mass loss in the temperature range 73 up to 236 ºC corresponding 22% weight loss is The thermal behaviour of Ag-OPTS associated with an endothermic heat effect. The complex was started in the temperature range observed mass loss (20%) corresponding from room temperature up to 800ºC under exothermic heat effect is connected with nitrogen flow. The corresponding TG and DTG thermolysis of the complex remaining[31]. The curves are presented in Fig. (9), showing three last step starts immediately after second step stages of mass loss. The first mass loss started and continue until complete decomposition of from room temperature with the removal of the ligand and formation of the end product as 2(HNCSNHNH) ended at 70◦C[30] and is silver metal. The decomposition of the complex associated with an endothermic heat effect. The was completed at ≥600ºC.

-37-

Fig. 9: Thermal analysis data and DTA of the Ag-OPTS complex

Fig. 10: ORTEP drawing of Ag-OPTS binary complex, showing their molecular structure and the labeling schemes used

CONCLUSION: novel bis(Schiff base) (2E,2'Z)-2,2'-(3-((E)-(2- hydroxyphenyl)diazenyl)pentane-2,4- diylidene) The straight forward condensation of o- bis(hydrazinecarbothioamide), the compound is hydroxyphenyl azo acetyl acetone and interesting in analytical chemistry and biology. thiosemicarbazide in 1:2 molar ratio yield the

-38- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Its composition and structure were determined 5-M. J. M. Campbell, Coord. Chem. Rev., 15 by elemental analysis and IR spectrometry. Its (1975) 279. flexible back bone, with the presence of N and S 6-J. S. Casas, M .S. Garcia-Tasende, J. Sordo, donor atoms, renders this compound interesting Coord. Chem. Rev., 209 (2000) 197. for studying its coordination behavior with 7-F. J. Giles, P. M. Fracasso, H. M. Kantarjian, metal ions (Fig. 10). The compound presents in J. E. Cortes, R. A. Brown, S. Verstovsek, Y. thione/thiol or both the tautomers forms. The Alvarado, D. A. Thomas, S. Faderl, G. interaction of OPTS with silver(I) leads to Garcia-Manero, et al., Leukemia Res. 27 formation of one different type of complex. In (2003) 1077. this work, the dissociation constants of the 8-G. Domagk, R. Behnisch, F. Mietzsch, H. reagent and the stability constants of their silver Smidt, Naturwissenschaftenm, 33 (1946) 315. complex were etermined by pH_metric and 9-A. I. Vogel (Practical Organic Chemistry) 3nd spectrometric titrations. The spectral data edn., p. 998, Longmans London (1974). indicate that when equimolar amounts of ligand 10-M. H. Moustafa; M. Abd-Almottaleb, T. A. and silver are used, the ligand can behave as a Seaf El-Naser, R. M. Issa and A. M. A. Awad; neutral tetradentate ligand to coordinate a Al-Azhar Bull. Sci., 15(1) (2004) 243. single metal ion through the C=N and C-SH 11-H. Irving and H. S. Rossotti; J. Chem. Soc.; groups. The coordinating behaviour of (1954) 2904. bis(thiosemicarbazone) ligand establishes that it 12-M.B. Ferrari, S. Capacchi, F. Bisceglie, G. is the azomethine nitrogen and thiol sulphur Pelosi, P. Tarasconi, Inorg. Chim. Acta, 312 which functions as the donor sites toward metal (2001) 81. ion. 13-A. Castineiras, E. Bermejo, J. Valdes- Martinez, G. Espinosa-Perez, D. X. West, J. REFERENCES: Mol. Struct. 522 (2000) 271. 14-V. Kuban and J. Havel; Acta Chem. Scand., 1-N. N. Gulerman, S. Rollas, H. Erdeniz, M. 27 (1973) 528. Kiraj, J. Pharm. Sci., 26 (2001) 1. 15-J. P. Phillips and L. L. Merrit, J. Am. Chem. 2-D. L. Klayman, J. F. Bartosevich, T. S. Soc. 70 (1948) 410. Griffin, C. J. Mason, J. P. Scovill, J. Med. 16-L. Sommer, Folia Fac. Sci. Nat. Uni. Purk., Chem., 22 (1979) 855. Brno, 5 (1964) 1. 3-D. L. Klayman, J. P. Scovill, J. F. Bartosevich, 17-P. Gans., A. Sabatini. and A. Vacca.; J. C. J. Mason, J. Med. Chem., 22 (1979) 1367. Chem. Soc., Dalton Trans., (1985) 1195. 4-P. Tarasconi, S. Capacchi, G. Pelosi, M. 18-M. H. Moustafa, Al-Azhar Bull. Sci. Cornia, R. Albertini, A. Bonati, P.P. Dall, P. (AISC'08), (March, (2008), 71. Luunghi, S. Pinelli, Bioorg., Med. Chem., 8 (2000) 154.

-39- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

19-M. H. Moustafa, Ass. Univ. Bull. Environ. 25- P. Job; Ann. Chem., 9 (1928) 113. Res.; 13, No. 2 (2010) 77. 26-F. G. Shirif and A. M. Awad, Inorg. Nucl. 20-M. H. Moustafa, Al-Azhar Bull. Sci., 12(2) Chem., 24 (1962) 79. (2010) 295. 27-J. H. Yoe and H. L. Jones, Indian Eng. 21-A. I. Matesanz, J. M. Pe´rez, P. Navarroa, J. Chem., Anal. Ed., 16 (1944) 111. M. Moreno, E. Colacio and P. Souza , J. 28-E. P. Sandell; Colorimetric Determination of Inorg. Biochem., 76 (1999) 29. trace metals, Vol. 3, Wiley, New York, (1959) 22-L. Sommer, V. Kuban and J. Havel; Folia p. 97. Fac. Sci. Univer. Brno, Vol, XI, (1970) pt. 1, 29-M. K. Das, M. Nath, J. J. Zuckerman, Inorg. 33. Chem. Acta, 21 (1983) 49. 23-P. Vozinca, J. Havel and L. Sommer; Collec. 30-N. M. El-Metwally, R.M. El-Shazly, I.M. Czech. Chem. Comm., 45 (1980) 54. Gabr, A.A. El-Asmy, Spectrochim. Acta Part 24-M. S. Abu-Baker, H. M. Rageh, E.Y. A, 61 (2005) 1113. Hashem and M. H. Moustafa; Monatsh. 31-K.-K. Du, S.-X. Liu, J. Mol. Struct., 874 Chemi., 125 (1994) 1197. (2008) 138.

تحضير ودراسات جهدية وطيفية على مركب بيس )ثيوسيميكاربازون( الجديد ومتراكبه مع أيون الفضة

محمود حسن مصطفى، أحمد عبد النعيم، أسامة أحمد عباس قسم الكيمياء - كلية العلوم للبنين – جامعة األزهر بأسيوط

تم فى هذا البحث تحضير قاعدة شيف الجديدة )E2 و E((-3(-′2,2- )Z ′2(-)2-هيدروكسى فينيل( داى ازينايل( بيتان-4,2-داى يلدين( بس )هيدرازين كربوثيواميد( المشتقة من الثيوسيميكربازيد. وذلك بتفاعل جزيئين ثيوسيميكربازيد مع مركب أرثو هيدروكسى فنيل أزو اسيتيل أسيتون، وتم اثبات التركيب الجزيئى لليجند بواسطة التحليل العنصرى وطيف الكتلة وطيف الرنين النووى المغناطيسى واألشعة تحت الحمراء, واستخدم هذه الليجئد لتحضير متراكب أيون الفضة. وتمت دراسة الخواص الحامضية والقاعدية للكاشف المحضر، وكذلك متراكب الفضة المحضر منه بواسطة الطرق الجهدية والطيفية المختلفة، وذلك بهدف تحديد حاالت اتزان التراكب الممكن تواجدها بالمحلول ونسب تكوينها وتدرج ثبات نظم التراكب المختلفة على ضوء طبيعة الكاشف الداخل فى التفاعل، وحسب ثوابت ثبات هذه النظم التركيبية المختلفة فى المعايرات عند تغير درجة تركيز أيون الهيدروجين بالمحلول وذلك لمعرفة الظروف المثلى لتكوين المتراكبات موضع الدراسة وكذلك استخدمت فى هذه الدراسات قياس أطياف االمتصاص المرئى والفوق بنفسجية خالل معايرات تتبع تغير درجة تركيز أيون الهيدروجين بالمحلول عند ظروف تجريبية محددة. وأمكن حساب ثوابت التأين لليجند وتحديد النسب التكوينية للمتراكب موضع الدراسة ووجدت بنسبة1:1. وقد أمكن تحضير المت راكب الصلب للفضة، وتم دراسة وتحديد مواضع الترابط مع أيون الفضة وتم التعرف على شكل وتركيب المتراكب.

-40- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

-2- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

EFFECT OF VITAMINS ON GROWTH CRITERIA, PHOTOSYNTHETIC PIGMENTS AND SOME METABOLIC PRODUCTS OF COBALTE CHLORIDE STRESSED SCENEDESMUS OBLIQUUS CULTURES Desouky, S.A., USAMA, M. A. and AHMED, W. A. Botany and Microbiology Department, Al-Azhar University, Faculty of Science, Assiut, 71524

ABSTRACT: This study show the effect of applied vitamins ascorbic acid (vitamin C) and thiamine (vitamin

B1) on growth criteria, total photosynthetic pigments total, carbohydrate, total protein, free amino acids and proline contents of stressed Scenedesmus obliquus cultured for 7 days. The growth criteria (cell count and dry weight), total photosynthetic pigments, total carbohydrate contents, total protein contents, free amino acids and proline of stressed Scenedesmus obliquus cultures were significantly increased, when the algal cultures subjected to lower doses (1.5 and 3 ppm) of

CoCl2 only. On the other side, under relatively higher concentration (4.5 ppm) of CoCl2 the cell count, dry weight, total photosynthetic pigments, total carbohydrate contents, total protein

contents and free amino acids of CoCl2 stressed Scenedesmus obliquus cultures were significantly decreased. However, the soluble carbohydrate contents, soluble proteins and praline contents of stressed Scenedesmus obliquus cultures were significantly increased. On the other hand, the parameters tested were significantly increased, when the algal cultures

were subject to various doses (1.5, 3 and 4.5 ppm) of CoCl2 and treated with 200 ppm applied vitamins (ascorbic acid or thiamine). However, the different carbohydrate fractions (soluble,

insoluble and total carbohydrate), soluble proteins of CoCl2 stressed Scenedesmus obliquus cultures were significantly decreased.

INTRODUCTION: shown to enhance growth of some plants at low concentrations. In higher concentration, cobalt It well known that the pollutions of fresh is toxic to human, terrestrial and aquatic water by heavy metals are considered the main animals and to plants. In this respect, cobalt is problem in the development countries. Cobalt is one from heavy metals group, which play an one of these elements, which cause the water important role in the growth parameters and pollution. Cobalt is an essential element for the metabolic activities of higher plants and algae. growth of marine algal species, including In this regards, Co+2 constituting 4% of vitamin diatoms, chrysophytes, and dinoflagellates B , appears to be universally essentially within (Bruland et al., 1991). Cobalt has also been 12 the plant kingdom as a micronutrient. It's only

-41- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

known biological function is associated with this 10-8 and 10-9 M of some heavy (Co+2, Cu+2 and vitamin; although organic compounds Zn+2) cause a clear reduction in the pigments containing Co+2 are know which reversibly bind content. Generally, the growth rates decreased oxygen (Wang and Schaeffer 1969 and Bunt in respect of increasing metals concentration 1970). Cobalt also activates several enzymes, more than 10-9 M of these heavy metals. especially those which are necessary for The total protein contents of the green alga metabolic process (Bowen, 1966 and Skholnik Chlorella vulgaris gradually decreased in a 1974). manner dependent on the metal concentration Cobalt has been found to inhibit in the medium (Afkar et al., 2010). On the other transmembrane ion movement and cell division hand, the supplementation of copper and zinc (Hagiwara et al. 1969). Ultra-structurally, Co+2 by concentration 10-9 M increases the total caused enlargement of the cell vacuole protein content as compared to the control. (Blankenship and Wilbur (1975). Also, Kumar However, no marked change in total protein and Kumar (1985) found growth inhibition of a contents occurred in cells of Chlorella vulgaris blue green alga Nostoc linkia to be directly which exposed to cobalt, Osman et al., (2004). proportional to increase concentration of cobalt The total free amino acids of Chlorella chloride. Also, Agrawal and kumar (1977) vulgaris gradually increased with increasing of studied the response of a blue green alga heavy metals concentration (Afkar et al., 2010). Anacystis nidulans to cobalt. They found that no The same authors found that, the most significant growth inhibition at the lower pronounced stimulation was detected at the concentration of (58.9 µg L-1) of Co+2, while, the culture supplemented with 10-7 M copper in growth inhibition was recorded at higher level comparison to the other tested metals. On the (589 µg L-1) of Co+2, and cell division stopped at other hand, cobalt and zinc also stimulated the concentration (5.9 mg L-1) of Co+2. In this biosynthesis of the total free amino acids, but context, Rachlin et al. (1982) found that, cobalt the stimulatory effect is less than that obtained caused significant decreased in cell size. with copper). It is well known that algal cells exposed to Proline accumulation under heavy metal heavy metals may suffer serious morphological stress has also been reported earlier in some and biochemical alterations (Rocchetta et al., higher plants (Bassi and Sharma, 1993), 2006). In this context, Afkar et al., (2010) found although the mechanism of accumulation of that the pigment contents of Chlorella vulgaris proline in plants or plant parts exposed to gradually increased, when the algal cultures stress. Thus, Maggio et al., (2002) found that the were subjected to low concentration 10-9 M of depicted an inverse relationship between +2 +2 +2 some heavy metals (Co , Cu and Zn ) during biomass and proline accumulation in the test exposure periods, whereas other concentrations algae under stressed conditions.

-42- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

The vitamin is an organic compound medium for enrichment and growth of the required as a nutrient in tiny amounts by an tested alga, (Stein, 1966). organism Lieberman and Bruning (1990). Treatments: Vitamins have diverse biochemical functions. Scenedesmus obliquus cultures subjected to Some have hormone-like functions as regulators 00 (control) and 200 ppm of ascorbic acid of mineral metabolism (e.g., vitamin D), or (vitamin C) and thiamine (vitamin B ) in the regulators of cell and tissue growth and 1 absence or presence of different levels (0, 1.5, 3, differentiation (e.g., some forms of vitamin A). and 4.5 ppm) of cobalt chloride for 7 days Others function as antioxidants (e.g., vitamin E incubations. and sometimes vitamin C) Bender (2003). The largest number of vitamins (e.g., B complex ANALYTICAL METHODS: vitamins) functions as precursors for enzyme 1-Determination of cell number: cofactors that help enzymes in their work as catalysts in metabolism. In this context, One drop of the algal suspension was Desouky (2008) working with Scenedesmus pipette on the Haemocytometer (0.1 mm depth), obliquus cultures subjected to 200 and 400 mM covered and left two minutes for algal setting. NaCl and treatment with 100 and 200 ppm The mean counts of four replicates were taken Salicylic acid (vitamin S) found that the growth into consideration and the results measured as -1 criteria (cell number and dry weight), total cells ml algal suspension. photosynthetic pigments, photosynthesis, 2-Determination of dry weight: respiration, proline, free amino acid and A definite volume (100 ml.) of algal catalase enzymes were significantly increased. suspension was filtered through weighed glass This study aim to elucidated the role of fiber filter. The cells after being precipitated on some external addition of vitamins (ascorbic the filter were washed twice with distilled water acid "vitamin C" and thiamine "vitamin B1") to and dried over night in an oven at 105 oC. The alleviate the adverse effect of CoCl2 on the data were expressed as µg 100 ml-1 algal growth criteria, total photosynthetic pigments suspension. and some metabolic activities of Scenedesmus obliquus cultures. 3-Determination of total photosynthetic pigments: MATERIALS AND METHODS: The pigment fractions (µg m1-1 algal Tested alga: suspension) chlorophyll a, chlorophyll b and Scenedesmus obliquus was collected from carotenoids were calculated by using the the River Nile and used as a test organism. equations mentioned by Metzner et al., (1965).

Beijerinck’s nutritive culture was used as a - Chlorophyll a = 10.3E663 - 0.918E664

-43- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

- Chlorophyll b = 19.7E664 - 3.87E663

- Carotenoids = 4.2 E452-(0.0264 chloro. a + 0.426 chloro. b)

-44- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

4-Determination of carbohydrate Whereas, the minimum values of growth contents: parameters were significantly decreased, when

Carbohydrate contents were determined algal cultures subjected to 4.5 ppm CoCl2 only, according to Badour (1959). (Fig. 1-a&b). The growth parameters and total 5-Determination of protein contents: photosynthetic pigments were significantly Protein contents were estimated according increased when algal cultures subjected to the method by Lowry et al., (1951). vitamins (200 ppm of either ascorbic acid

"vitamin C" or thiamine "vitamin B1"). This 6- Determination of proline: increase depending on concentration and level It was determined according to Bates et al, of CoCl2 and type of applied vitamin. (1973) methods. Ascorbic acid treatments to non-polluted

7- Determination of free amino acids: Scenedesmus cultures, the contents of cell number, dry weight and total photosynthetic Free amino acids were extracted from fresh pigments were raised (Fig. 1-a). On the other water algal suspension and calorimetrically side, the contents of cell number, dry weight and determined using the method of Moore and total photosynthetic pigments were raised, when Stein (1948). treated with thiamine, (Fig. 1-c). 8- Statistical Analysis: The data were statistically analyzed to Metabolic products: calculate the Least Significant Difference The various contents of carbohydrate (L.S.D) according to Snedecor and Cochran fractions under different levels (1.5, 3 and 4.5

(1980). ppm) of CoCl2 exhibited higher contents than those of the control cultures. However, highest

RESULTS: level of CoCl2 (4.5 ppm) lowered in most cases The growth criteria (cell count and dry the content of these carbohydrate fractions. weight) and total photosynthetic pigments of The maximum value of soluble Scenedesmus cultures were increased up to the carbohydrates content amounted t0 130% of level 3 ppm of CoCl2. However, under relatively that of the control cultures, when algal cultures higher level (4.5 ppm) of CoCl2, all these subjected to 3 ppm CoCl2. Also, the maximum parameters were decreased as compared with values of insoluble and total carbohydrates that of the control cultures. content amounted to 139% and 134% of that of The maximum values of growth parameters the control cultures, when algal cultures and total photosynthetic pigments of subjected to 1.5 ppm CoCl2, respectively. But, Scenedesmus were significantly increased, when the minimum values of soluble, insoluble and algal cultures subjected to 3 ppm CoCl2 only, total carbohydrates content amounted to 92%,

-45- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

93% and 93% of that of the control cultures, content were reported, when the algal cultures when algal cultures subjected to 4.5 ppm CoCl2, subjected to 3 ppm CoCl2 and treated with respectively (Fig. 2-a). thiamine, respectively (Fig. 2-c). Addition 200 ppm of either ascorbic acid or The maximum value of soluble protein thiamine to different levels of CoCl2 changed the contents when treated with CoCl2 amounted to content of carbohydrate fractions (soluble, 133%, that of the control cultures when algal insoluble and total carbohydrates). cultures subjected to 1.5 ppm CoCl2. Whereas, When, the non-polluted Scenedesmus the maximum values of insoluble and total obliquus cultures treated with vitamins the protein contents when treated with CoCl2 only content of soluble, insoluble and total amounted to 132% and 125% of that of the carbohydrates were raised and amounted to control cultures, when algal cultures subjected

185%, 142% and 149% that of the control to 3 ppm CoCl2, respectively. In contrast, the cultures, when the algal cultures treated with minimum values of soluble, insoluble and total ascorbic acid, respectively (Fig. 2-b). Whereas, protein contents when treated with CoCl2 only addition of thiamine to non-polluted amounted to 81%, 67% and 70% of that of the Scenedesmus cultures significantly increased the control cultures when algal cultures subjected to content of soluble, insoluble and total 4.5 ppm CoCl2, respectively (Table 1-a). carbohydrates (Fig. 2-c). Treating of non-polluted Scenedesmus The maximum value of soluble cultures with 200 ppm of either ascorbic acid or carbohydrates content amounted to 92% of that thiamine resulted in promotion of soluble, the control cultures, when the Scenedesmus insoluble and total protein fractions giving cultures subjected to 1.5 ppm CoCl2 and treated amounts of 160%, (Table 1-b). While, it reached with 200 ppm of ascorbic acid respectively. the to 152%, 151% and 151% of that of the control maximum values of insoluble and total cultures when algal cultures treated with carbohydrates content were 105% and 99% as thiamine, respectively (Table 1-c). compared with that of the control cultures, The results given in table (2) indicate that when the algal cultures subjected to 3 ppm heavy metals exerted generally a stimulatory

CoCl2 and treatment with ascorbic acid, effect on the accumulation of proline in the test respectively (Fig. 2-b). algal cultures. The highest proline level was On the other side, the maximum value of consistently found in cultures incubated under soluble carbohydrates content amounted to the highest concentration of heavy metal used. 85% that of the control cultures, when the algal The maximum value of proline cultures treated with thiamine and subjected to accumulation amounted to 574%, that of the

1.5 ppm CoCl2, respectively. Also, the maximum control cultures, when algal cultures subjected values of insoluble and total carbohydrates to 4.5 ppm CoCl2 (Table 2-a).

-46- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 1: Protein contents (µg mg-1 dry weight) of Scenedesmus obliquus cultures subjected to various combinations of CoCl2 and 200 ppm of either ascorbic acid (vitamin C) or thiamine (vitamin B1) for 7 days incubation Treatments Water-soluble % Water-insoluble % Total %

Proteins Control Proteins Control Proteins Control CoCl2 (ppm) (a) CoCl2 00:00 33.00 100.00 142.00 100.00 175.00 100.00 1.5:00 44.20** 133.94 167.50 ** 117.96 211.70** 121.00 3:00 31.10** 94.24 188.25** 132.60 219.35** 125.24 4.5:00 26.80** 81.21 96.00** 67.61 122.80** 70.17 L.S.D at 1 % 1.002 8.002 12.005

L.S.D at 5 % 2.004 10.045 24.008

(b) Ascorbic acid (vitamin C) CoCl (ppm) &Ascorbic acid (ppm) 2 00 : 200 53.05** 160.76 230.00** 162.00 283.05** 161.74 1.5:200 60.81** 180.30 259.37** 182.65 320.18** 182.96 3:200 66.00** 200.00 283.40** 199.30 349.40** 199.66 4.5:200 55.53** 168.30 205.00** 144.40 260.53** 148.87 L.S.D at 1 % 7.002 14.002 15.003

L.S.D at 5 % 8.004 20.004 22.007

(c) Thiamine (vitamin B1) CoCl (ppm) & Thiamine (ppm) 2 00:200 50.43** 152.82 215.00** 151.40 265.43** 151.70 1.5:200 61.00** 184.85 250.37** 176.32 311.37** 178.00 3:200 63.42** 192.18 283.44** 199.60 346.86** 198.21 4.5:200 55.50** 168.18 194.76 137.15 250.26** 143.01 L.S.D at 1 % 7.025 20.124 11.123

L.S.D at 5 % 9.125 25.456 15.123

Table 2: Proline contents and free amino acids (µg mg-1 dry weight) of Scenedesmus obliquus cultures subjected to various combinations of CoCl2 and 200 ppm of either ascorbic acid (vitamin C) or thiamine (vitamin B1) for 7 days incubation Treatments % % Proline Free amino acids Control Control CoCl2 (ppm) 00:00 0.93 100:00 10.88 100 1.5:00 1.88** 202.15 12.78** 117.46 3:00 3.45 371.06 17.32** 159.20 4.5:00 5.34** 574.20 10.00 92.00 L.S.D at 1 % 0.001 0.008

L.S.D at 5 % 0.004 0.008

2-a: Thiamine (vitamin B1) CoCl2 (ppm) &Ascorbic acid (ppm) 00:00 1.36** 146.24 23.35** 214.60 1.5:00 1.80** 193.55 31.52.92 293.40 3:00 1.15** 123.65 38.91** 357.63 4.5:00 1.84** 197.85 2.04** 239.34 L.S.D at 1 % 0.005 8.124

L.S.D at 5 % 0.007 10.450

2-b: Thiamine (vitamin B1) CoCl2 (ppm) & Thiamine (ppm) 00:00 2.82** 303.23 38.00** 349.36 1.5:00 1.75** 188.20 26.51** 234.66

-47- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

3:00 1.56** 167.74 35.82 329.23 4.5:00 2.67** 287.10 21.61** 198.62 L.S.D at 1 % 0.057 9.245

L.S.D at 5 % 0.054 11.456

* Significant difference ** High Significant difference…………… as compared absolute control

-48- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Cell count Fig.Fig 1. -(a1 ) 180 Dry weight Total pigments 160 140 120 100 80

%Control 60 40 20 0 00:00 1.5:00 3:00 4.5:00 Treatments : CoCl2 ppm

Fig.Fig. (11--ba ) Cell count 300 Dry weight Total pigments 250 200 150

%Control 100 50 0 00:200 1.5:200 3:200 4.5:200

Treatments : CoCl2 ppm and 200 ppm ascorbic acid

Fig.Fig. 1(1-c-b ) Cell count 250 Dry weight Total pigments 200

150

100 %Control 50 0 00:200 1.5:200 3:200 4.5:200

Treatments :CoCl2 ppm and 200 ppm thiamine

Figure 1: Cell count (cell ml-1 algal suspension), dry weight (µg ml-1 algal suspension) and total photosynthetic -1 pigments (µg ml algal suspension) of Scenedesmus obliquus cultures subjected to various combinations of CoCl2 and 200 ppm of either ascorbic acid (vitamin C) or thiamine (vitamin B1) for 7 days incubation

-49- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Fig. (2) Soluble carbohydrates 160 Fig. 2-a Insoluble carbohydrates 140 Total carbohydrates 120 100 80

%Control 60 40 20 0 00:00 1.5:00 3:00 4.5:00 Treatments : CoCl2 ppm

200 Fig. (2-a) Soluble carbohydrates Fig. 2-b Insoluble carbohydrates Total carbohydrates 150

100

%Control 50

0 00:200 1.5:200 3:200 4.5:200

Treatments : CoCl2 ppm and 200 ppm ascorbic acid

Fig. (2-b) Soluble carbohydrates 200 Fig. 2-c Insoluble carbohydrates Total carbohydrates 150

100 %Control 50

0 00:200 1.5:200 3:200 4.5:200

Treatments : CoCl2 ppm and 200 ppm thiamine

Figure (2): Carbohydrate contents (µg mg-1 dry weight), of Scenedesmus obliquus cultures subjected to various concentrations of CoCl2 and 200 ppm of either ascorbic (vitamin C) acid or thiamine(vitamin B1) for 7 days incubation

-50- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

The highest proline content of Scenedesmus polluted algal cells by heavy metals is also obliquus recorded at the level of 4.5 ppm CoCl2 expressed in changes in the biosynthesis of free only was associated with a significantly amino acids. These changes are given in table decreased in the contents of free amino acids (2). The inhibitory effect of heavy metals on the and thus the lower value was 92% of that of the rat of biosynthesis of free amino acids than control cultures, when algal cultures subjected proline considerably varied. to 4.5 ppm CoCl2 only, whereas highest value of The highest proline content of Scenedesmus amino acids content was 159% in algal cultures obliquus recorded at the level of 4.5 ppm CoCl2 subjected to 3 ppm CoCl2 (Table 2-a). only was associated with a significantly Using of either ascorbic acid or thiamine as decreased in the contents of free amino acids additional treatments to non-polluted and thus the lower value was 92% of that of the Scenedesmus cultures resulted in an increase in control cultures, when algal cultures subjected the content of free amino acids. This content to 4.5 ppm CoCl2 only, whereas highest value of amounted to 214% and 349% of that of the amino acids content was 159% when algal control cultures when algal cultures treated cultures subjected to 3 ppm CoCl2 only (Table 2- with 200 ppm of either ascorbic acid or a). thiamine, respectively (Table 2-b&c). Using 200 ppm of either ascorbic acid or The maximum value of proline thiamine as additional treatments to non- accumulation amounted to 574%, of that of the polluted Scenedesmus cultures resulted in control cultures, when algal cultures subjected increasing the content of free amino acids. This to 4.5 ppm CoCl2 only (Table 2-a). In this content amounted to 214% and 349% of that of context, in polluted-Scenedesmus cultures, the the control cultures when algal cultures treated maximum value of proline content were 287% with 200 ppm of either ascorbic acid or of that of the control cultures, when algal thiamine, respectively (Table 2-b &c). cultures subjected to 4.5 ppm CoCl2 and treated with 200 ppm of either ascorbic acid or DISCUSSION: thiamine, respectively. Also, the minimum Adverse effect of heavy metals on green values of proline contents were 177% of that the algae may lead to disturbances in plant control cultures, when the algal cultures metabolism which consequently lead to a subjected to 1.5 ppm CoCl2 and treated with 200 reduction in the growth of these algae (Desouky, ppm of either ascorbic acid or thiamine (Table "1995, 2001, 2003 and 2004" and Afkar et al., 2-b &c). 2010). The important role played by applied The results in this study showed the growth vitamins (200 ppm of either ascorbic acid parameters and total carbohydrate contents of

"vitamin C" or thiamine "vitamin B1") in Scenedesmus obliquus cultures were

-51- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

significantly increased, when the algal cultures doses. On the other hand, the supplementation subjected to lower level (1.5 and 3 ppm) of of copper and zinc by concentration 10-9 M

CoCl2. While under higher dose of the increases the total protein content as compared carbohydrate contents were significantly to the control. However, no marked change in decreased. However, the soluble carbohydrate total protein contents occurred in cells of contents were increased with the increasing of Chlorella vulgaris which exposed to cobalt. It

CoCl2 levels in the medium cultures. These could be suggested that accumulation of protein results present in this study are in agreement at low heavy metal concentrations may be one of with Afkar et al., (2010) that repotted the the ways through which the algae can abolish carbohydrate contents of the Chlorella vulgaris their toxic effects, or to increase respiration alga also declined in manner dependent on the leading to the utilization of carbohydrate in metal concentration exist in the medium. In this favor of protein accumulation (Osman et al., context, Fathi et al. (2005) reported that the 2004). These results in this investigation in higher doses of heavy metals have severely accordance with Afkar et al., (2010) reported attenuate chlorophyll synthesis coupled with that the total free amino acids of Chlorella severe drop in protein resulting in increased vulgaris gradually increased with increasing carbohydrates. In this respect, Desouky, (2004) metals concentration. On the other hand, cobalt found that the total carbohydrate contents of and zinc also stimulated the biosynthesis of the Chlorella vulgaris cultures were significantly total free amino acids, but the stimulatory effect decreased when; the algal cultures were is less than that obtained with copper. subjected to various concentrations (5, 10, 20, 25 Generally, the accumulation of amino acids and 50 ppm) of CoCl2. in response to metals concentrations may lead to The results in this study showed the protein the assumption that suppressed protein contents of Scenedesmus obliquus cultures were biosynthesis encouraged free amino acids significantly increased, when the algal cultures accumulation, or may be due to some subjected to lower levels (1.5 and 3 ppm) of counteracting chelating mechanism against . CoCl2, only However under higher doses of (4.5 heavy metals toxicity (El-Sheekh et al., 2003, ppm) of CoCl2 the protein contents decreased. Osman et al., 2004 and Fathi et al., 2005). The soluble protein contents increased with the Also, the results in this study showed the increased of treatment element in the medium proline contents of Scenedesmus obliquus cultures. Under higher concentrations of this cultures were significantly increased, when the element (Co+), the total protein contents algal cultures subjected to low levels (1.5 and 3 decreased, when compared with that the control ppm) CoCl2. While under higher level (4.5 ppm) cultures. The same authors reported that all the of CoCl2, the proline contents were significantly three metals (Co+2, Cu+2 and Zn+2) affected increased, under heavy metal stress has also negatively the total protein contents at higher been reported earlier in some higher plants

-52- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

(Bassi and Sharma, 1993), although the and Makled, 1995). In this context, Desouky mechanism of accumulation of proline in plants (2001) found that the growth parameter of or plant parts exposed to stress is still unknown. Chlorella vulgaris cultures (cell number and dry Vitamins are nutritional organic weight), total photosynthetic pigments and some compounds required for continued growth by metabolic activities increased when the algal phytoplankton species when studied in axinic cultures were subjected to 200 ppm applied cultures. These vitamins are required in very vitamin (thiamine). low concentrations Bender (2003), as cofactors In this investigation, total carbohydrate by organisms which undergo autotrophic contents of Scenedesmus obliquus cultures were growth (Berland et al, 1978, Swift, 1980 and significantly decreased when the algal cultures

Desouky, 2011). subjected to various doses of CoCl2 and Plants generally synthesize large amounts of treatment with any vitamins (200 ppm of either ascorbic acid which could be utilized in cell ascorbic acid or thiamine). These results in this metabolism (Arrigoni et al, 1975 and Chinoy, study in accordance with, Desouky, (2003 and 1984), and could activate the biological defense 2004) recoded that the total carbohydrate mechanisms (Arrigoni et al., 1979). contents of Chlorella vulgaris cultures were In this investigation, it was generally found significantly decreased, when the algal cultures that the concentrations of the applied vitamins were subjected to various levels of either salinity (200 ppm ascorbic acid "vitamin C" or or heavy metals and treatments with any thiamine "(vitamin (B1") were significantly vitamins (200 ppm of ascorbic acid or thiamine). increased, the growth parameters (cell number The counteractive effects of thiamine might and dry weight) and total photosynthetic link with the physiological role of this vitamin. pigments of Scenedesmus obliquus cultures, Such vitamins can be used as coenzyme when subjected to various doses (1.5, 3 and 4.5 performed generally the major physiological ppm) of CoCl2. function in the plant cells (Kefeli, 1981). Ascorbic acid was also recorded to affect the Mohamed et al., (1988) found that thiamine chlorophyll contents (Choudhury et al., 1993) increased the protein contents of Chlorella through promoting the capacity of chlorophyll by vulgaris, and the highest protein contents were stabilizing and protecting these molecules from produced in the presence of 100 ppm being oxidized. This stimulation of growth of concentration of thiamine. Similarly, Makled both organisms mentioned above in response to (1995) working with two green algae namely treatments with thiamine (vitamin B1) could be Chlorella vulgaris and Ankistrodesmus falcatus, attributed to the significant role played by reported that the addition of thiamine, nicotinic thiamine (vitamin B1) in cellular metabolism as acid or pyridoxine exerted considerable increase coenzymes in the oxidative decarboxylation of in growth rate, synthesis of total photosynthetic pyruvate or of α-ketoglutarate (Harper, 1991 pigments, carbohydrate and protein contents.

-53- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

In this study, total protein content of ascrobic acid biosyntesis. Nature, 256: Scenedesmus obliquus cultures were increased, 746-756. when the algal cultures were subject CoCl2 and Arrigoni, O., De-Gara, L. Tommasi, F. and treated with any applied vitamins (200 ppm Liso, R. (1992): Changes in the asorbate ascorbic acid or thiamine). In this respect, system during seed developments in Vicia Mohamed et al. (1988) working with Chlorella faba L. Plant Physiol. 59:335-338. vulgaris, found that thiamine (Vitamin B1) Arrigoni, O., Zacheo, G., Arrigoni_Lisio, R. and increased the protein contents, and the highest Bleve_Zacheo, T. (1979): Relationship protein content was produced in the presence of between Ascorbic acid resistance in -4 10 M thiamine (Vitamin B1). tomato plants to Meloidogyne incognita. In this investigation, the free amino acid Phytopathology 69: 579-581. and proline contents of Scenedesmus obliquus Badour. S. S. A. (1959): Analytish-chemish cultures were significantly increased, when the untersuchung des kaliummangles bei algal cultures were subjected with various doses Chlorella im vergleich mit anderen

(1.5, 3 and 4.5 ppm) of CoCl2 and treated with Mangelzustän den, Ph. Dissertation. any applied vitamins (200 ppm ascorbic acid or Goettingen. thiamine). These results in this investigations in Bassi, R. and Sharma, S.S. (1993): Proline accordance with, some other authors working accumulation in wheat seedling exposed with some types plants found that the growth to zinc and copper. Phytochem. 33, 1339- parameters, total photosynthetic pigments, total 1342. carbohydrates, total protein free amino acid Bates, L.S.; Waldren, R.P. and Teare, I.D. and proline contents were markedly raised, (1973): Rapid determination of free when exposed either salinity stress or heavy proline for water stress studies. Short metals and treatment with any of the applied communication, Plant Soil 39: 205-207. vitamins (200 ppm of ascorbic acid, thiamine or Bender, A. (2003): Nutritional biochemistry of pyridoxine) (Arrigoni et al., 1992, Desouky, the vitamins. Cambridge, U.K.: 2001, 2003, 2004 and 2011). Cambridge University Press. Berland, B.R., Bonin, D. J., Fiala, M. and REFERENCES: Maestrin, S.Y. (1978): Importance des vitamins en mer. consommation et Afkar, E., H. Abanna and A.A. Fathi (2010): production par les algues et les bacteries, Toxicology response of green algae In: Actualites de biochimie marine. Chlorella vulgaris, to some heavy metals. colloque G. A. B. I. M-C. N. R. S, Amer. J. Enviro. Sci. 230-237. Marseille 1976 Ed. Special du C.N.R,S., Arrigoni, O. Arrigoni-Lisio, R. and Calabrese, Paris. G. (1975): Lycorine as an inhibitor of

-54- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Blankenship, M.L., and Wilbur, K.M. (1975): growth parameters, photosynthesis, Cobalt effect of cell division and calcium respiration, carbohydrates, proteins, free uptake in the coccolithophorid amino acids and proline of Chlorella Cricoshaera carterae (Haptophyceae), J. vulgaris Beijer cultures. Al-Azhar Bull. Phycol. 11, 211. Sci. Proceeding of the 5 th. Int. Sci. Conf, Bowen, H. J. M. (1966): Trace Elements in 25-27 March 2003. PP: 277-279. Biochemistry Academic Press, London. Desouky, S.A.(2001): Effect of various Bruland, K. W., Donat, J. R., and Hutchins, D. concentrations of zinc chloride upon A. (1991): Interactive influences of growth criteria, oxygen evolution and bioactive trace metals on biological oxygen uptake of Chlorella vulgaris production in oceanic waters. Lim. Ocen. Beijer. Ass. Univ. Environ.: 42-50. 36(8): 1555-1577. Desouky, S. A. (2004): Response of cadmium – Bunt, H.S., (1970): Uptake of cobalt and vitamin stressed Chlorella vulgaris Beijer cultures

B12 by tropical marine microalgae, J. to riboflavin (B2). The Sec. Int. Conf. for Phycol. 6, 339. Develop. In the Arabic world, March, 23- Chinoy, J. J. (1984): The role of ascorbic acid in 25, pp: 37-48. growth, differentiation and metabolism of Desouky S. A. (2008): Effect of salicylic acid on plants. Chnoy, N. J., Nijhoff, M. and Junk, photosynthesis and some antioxidant W. (ed). Publisher, the Hague and the enzymes of stressed Scenedesmus obliquus Netherlands, Chlorella vulgaris response culturess Al-Azhar Bull. Sci. Vol. 19: pp: of the antioxidant system. J. Plant 91-108. Physiol. 161, 591–599. Desouky, S. A. (2011): Effect of some natural Choudhury, N. K., Choe, H.T. and Huffeker, R. organic additives on the growth and C. (1993): Ascorbate induced zeaxanthin photosynthesis of pollutant-Chlorella formation in wheat leaves and vulgaris Beijer. J. Appl. Sci. Res. 7 (1) 23- photoproduction of pigment and 32. photochemical activities during aging of El-Sheekh, M. M., El-Naggar, A. H., Osman M. chloroplasts in light. J. Plant Physiol. 141: E. H. and El-Mazaly, E. (2003): Effect of 551-556. Cobalt on growth, pigments and the Desouky, S. A., (1995): Effect of some organic photosynthetic electron transport in additives on salinized Chlorella vulgaris. Monoraphidium minutum and Nitzchia Ph.D. Thesis, Fac. Sci., Assiut Univ., perminuta.Braz. J. Plant Physiol., 15: 159- Egypt pp: 1-199. 166. Desouky, S. A., (2003): Alleviation the toxicity Fathi, A. A., F. T. Zaki and H.A. Ibraheim, effect of lead acetate by riboflavin on (2005): Response of tolerant and wild

-55- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

type strains of Chlorella vulgaris to Mutanten Von Chlorella. Planta 65:186- Copper with special references to Copper 194. uptake system. Protistology, 4: 73-78. Mohamed, Y. A., Osman, M.E.H., and El- Hagiwara, S., Hayashi, H. and Takashashi, K. Sheekh, M. M. (1988): Factors affecting (1969): Calcium and potassium currents Growth and Protein Content Chlorella of the membrane and a barnacle muscle vulgaris. Bull. Fac. Sci. Assiut Univ., fiber in relation to the calcium spike, J. Egypt, 17: 103-114. Physiol. 205, 115. Moore, S. and W. Stein, (1948): Photometric Harper, H.A. (1991): The water soluble ninhydrine method for use in the vitamins. In: Review of physiological chromatography of amino acids. J. Biol. chemistry, Los Altos, California: 159-187. Chem., 17: 367-388. Kefeli, V. I. (1981): Vitamins and some other Osman, M.E.H., A.H. El-Naggar, M. M. El- representatives of nonhormonal plant Sheekh and E. El-Mazally, (2004): growth regulators. Priki Biokhim. Differential effects of Co+2 and Ni on Microbiol, 17: 5-15. protein metabolism in Scenedesmus Kumar, D., Jha, M. and Kumar, H. D. (1985): obliquus and Nitzschia perminuta. Heavy metal toxicity in the Environ. Toxicol. Pharmacol., 16: 169- cyanobacterium Nostoc linckia, Aquat. 178. Bot. 22, 101. Rachlin, J. W., Jensen, T.E., Baxter, M. and Lieberman, S., Bruning, N. (1990): The Real Jani, V. (1982): Utilization of Vitamin & Mineral Book. NY: Avery morphometric analysis in eveluting Group, 3. response of Plectonema boryanum Lowry, O. H., Rosebrough, N. J., Farr, A. L. (cyanophyceae) to expousure to eight and Randall, R. J., (1951): Protein heavy metals, Arch. Environ. Contam. measurement with the Folin phenol Toxicol. 11, 323. reagent. J. Biol. Chem. 193, 265–275. Rocchetta, I., V. Mazzuc and M.R. Carmen, Makled, M. M. (1995): Physiological studies on (2006): Effect of chromium on the fatty some species of unicellular green algae acid composition of two strains of with special reference to protein Euglena gracili. Environ. Pollut., 141: production. M. Sc. Thesis, Fac. Sci., 353-358. Menoufia Univ.., Egypt, pp.1-195. Skholnik, M. J. (1974): Micronutrients in the Metzener, H., Rau, H. und Senger, H. Plant Life, Nauka, Leningrad (in (1965): untersuchungen Zur Synchronisi- Russian). erbarkeit einzelener Pigment-Mangel

-56- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Snedecor, G.A. and W.G. Cochran, (1980): phytoplankton. Morris, I. (ed) Backwell Statistical Methods, 11th. Ed., the Iowa Scientific Publications, Oxford, London, State Univ. Press, Ames, Iowa, U. S. A., Edinburgh, Boston, 6 : 329-368 pp: 172-334. Wang. B. and Schaeffer, W. P. (1969): Structure Stein, J. R. (1966): Growth and mating of of an oxygen-carrying cobalt complex, Gonium pectoral (Volvolcales) in defined Science, 166, 1404. media. J. Phycol. 2: 23-28. Swift, D.G. (1980): Vitamins and phytoplankton growth. In: The physiological ecology of

-57- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

تأثير الفيتامينات علي النمو ، األصباغ النباتية وبعض المنتجات األيضية لمزارع طحلب "السينيدسمس أوبليكس"المجهدة بكلوريد الكوبلت سيد عباس دسوقي، أسامة محمد عبدالرؤوف، أحمد ورداني عبدالراضي جامعة األزهر ـ كلية العلوم ـ قسم النبات والميكروبيولوجي ـ أسيوط ـ 42517 البريد األلكتروني: [email protected]

اهتمت تلك الدراسة بإظهار تأثير الفيتاميناات ])حما اسساربربيك )فيتاامي ( بالثياامي )فيتاامي 1([ علا مدادست النمااب )عاادد اليايااا بالاابا ال ااا ،( األصااباا النباتيااة، محتاابا المااباد الررببهيدراتااة، محتاابا المااباد البربتينااة، األحمااا األمينية بالبربلي علي ماارع طحل " السينيدسمس أببليرس " الم هدة يرلبريد الرببلت لمدة سبدة أيام. بيمر تلييص النتائج التي أمر الحصبل عليها في هذه الدراسة، رما يلي : 1- عند مدال ة طحل ماارع طحلا " السينيدسامس أببلايرس الم هادة بترريااات ميتلفاة ما رلبرياد الرببلات ),،1، 3،0، ,،4( اء م المليب منفردا . سحظ أ مددل النمب متمثا في )عدد الياياا بالابا ال اا ( باألصاباا النباتياة المحتابا الرلااي لراال ماا المااباد الررببهيدراتيااة بالبربتينيااة باألحمااا األمينيااة بالبااربلي لماااارع لطحلاا " السينيدساامس أببلاايرس" الم هدة برلبريد الرببلت تاداد ايادة مدنبية حت مستبى 3 اء م المليب .

2- عنااد الترريااا الدااالي ),،4 اااء ماا المليااب ( تتنااا ص مداادست النمااب باألصااباا النباتيااة المحتاابا الرلااي لراال ماا المااباد الررببهيدراتية بالبربتينية باألحما األمينية المحتبا الرلي لرل م المباد الررببهيدراتية بالبربتينية تنا صا مدنبيا . بعلي الدرس م ذلك ياحظ ايادة محتبا المباد الررببهيدراتية بالبربتينية الذائبة بالبربلي .

3- اادياد المددست الميتبرة تاداد ايادة مدنبية، بذلك عند مدال ة ماارع طحل " السينيدسمس أببليرس " الم هدة بتررياات ميتلفة ),،1، 3،0، ,،4( اء م المليب م رلبريد الرببلت بباسطة 200 اء م المليب م أا م الفيتااميني . أمااا المحتاابا إلاا للمااباد الررببهيدراتيااة )الذائبااة برياار الذائبااة بالرليااة( بمحتابا الماباد البربتنياة الذائباة تتناا ص تنا صاا مدنبيا .

-58- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

ESTIMATION OF SENSIBLE HEAT LOSS IN CAPSAICIN- DESENSITIZED CHICKEN AFTER EXPOSURE TO DISRUPTION OF THERMAL HOMEOSTASIS Motamed Elsayed Mahmoudc, Reem Mahmoud Dosokyb, Moustafa Mohammed Ahmedb & Tadashi Takewakia a- Department of Basic Vet. Science, Laboratory of Physiology, the United Graduate School of Vet. Sciences, Gifu University, Japan. b- Department of Animal Hygiene, Faculty of Vet. Medicine, Assiut University, Egypt. c- Department of Animal Husbandry Faculty of Vet. Medicine, Sohag University, Egypt.

ABSTRACT:

-57- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Background: Thermal conditioning in newly hatched chicken is of great importance. Induction of thermotolerance to cold, heat and disrupted thermal balance is one of the most important managemental tools used to improve survival in chicken. It is well known that chicken is in sensitive to capsaicin (main ingredient of hot chilli peppers) due to insensitivity of Transient membrane potential of vanilloid subtype-1 (TRPV1) receptors to capsaicin. These receptors are responsible for perception of pain burning sensation and thermoregulation in mammals. Owing to the capsaicin's availability as an rodent repellent function in poultry ration and its preference by many birds, we investigated the mechanism of thermoregulation of capsaicin in chicken The purpose: To interpret the phenomenon of thermotolerance in capsaicin (CAP) desensitized chicken and to study its effect on sensible heat loss mechanisms in newly hatched chicks. Methods: In this study, chicken were treated intravenously (IV) in wing vein once with CAP (10 mg/kg, body weight) at 1st, 2nd & 3rd days of age. Then after one week from CAP- pretreated chicken were exposed to cold (8°C), heat (38°C) or injected with lipopolysacccharide (LPS; outer memebrane of Gram negative bacteria). LPS at a low dose (1 mg/kg, body weight, IV) induce fever or at a high dose (10 mg/kg, IV) to induce hypothermia. Surface (skin of back) and colonic temperatures were measured to calculate heat loss index (HLI) as an indicator to the sensible heat loss. Main results: The HLI was 0.95 in control non-treated chicken at ambient temperature (Ta 25°C). In CAP-desensitized chicken HLI was increased to a maximum of 0.97 at Ta 38°C and up to 0.99 at climax of fever induced by LPS. The controversial finding was observed in chicken exposed to cold; HLI in CAP-desensitized chicken was not increased but reduced to 0.94, however no such effect of capsaicin at nadir of hypothermia induced by high dose of LPS. Conclusion: CAP-sensitive receptor (Transient membrane potential of vanilloid subtype-1;TRPV1)- independent pathway may exert a thermoregulatory role during heat and cold exposure, and in LPS-induced fever in part through affecting sensible heat loss in chicken.

-58- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

INTRODUCTION: and free radicals like NO are released in Chicken as homeothermic species is able to animals (Kluger, 1991; Romanovsky et al., 1996, keep their body core temperature balanced 2005; Saia & Carnio, 2006 and Steiner et al., within a very narrow range (within 1°C) even 2006) and in chicken (Nikami et al., 2008). In when exposed to a wide range of ambient mammals, LPS induces hypothermia by temperatures (Romanovsky, 2007). At high reducing metabolic heat production and by an temperature, heat production decreases while induction of cold-seeking behavior, even though heat dissipation increases. The main pathway of body temperature is decreasing (Kluger, 1991; heat dissipation for birds under hot Steiner and Branco, 2002; Romanovsky et al., environment is via panting (Richard, 1971 and 2005; Rudaya et al., 2005 and Almeida et al., Shinder et al., 2007). Nonevaporative heat loss 2006a, b). However, the thermoeffector takes place at the surface of bare skin and mechanisms initiating cold- and high dose of plumage. The resultant level of skin LPS- induced hypothermia which seemed to temperature depends on the rate of heat loss oppose each other, and despite of the similar and the rate at which warm blood flows from responses of CAP-pretreated chicks to both core to the skin. Therefore, the skin inflammatory (LPS) and thermal (cold) stimuli, temperature could give a certain reflection of as one major difference. Exposure to cold may the response of thermoregulation (Nääs et al., activate autonomic and behavioral mechanisms 2010). to increase heat production and reduce heat loss Disruption of thermal balance can occur by at maxima, and body temperature drops when thermal stimuli such as heat and cold, and non- heat loss exceeds heat production. In contrast, thermal stimuli such as Lipopolysachharide LPS-induced hypothermia is accompanied by (LPS) injection. LPS causes fever or decrease of heat production and cold-seeking hypothermia depending on the dose of injection. behavior in animals (Romanovsky et al., 1996, The fact that CAP pretreatment attenuates both 2005; Mailman et al., 1999 and Almeida et al., LPS-induced fever (Mahmoud et al., 2007) and 2006a, b) and chicken (Mahmoud et al., 2007). LPS-induced hypothermia (Nikami et al., 2008) Therefore, the mechanisms responsible for as they suggested that the CAP-sensitive, prevention of cold- induced hypothermia by TRPV1-independent pathway was not only CAP-pretreatment would be independent of specifically involved in hyperthermic reactions those operating in the prevention of LPS- but also was commonly related to disruption of induced hypothermia. CAP pretreatment also thermoregulation. Both LPS induced fever and rescued cold-induced hypothermia (Nikami et hypothermia that arise from processing of LPS al., 2008). Although, CAP has wide rage of anti- by macrophages and endothelial cells in major inflammatory actions (Kim et al., 2003 and organs (liver, lung and brain), where cytokines Chen et al., 2003) this study was focused on the

-59- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011 effect of CAP of sensible heat loss that indicated 3-Experimental model of disrupted by heat loss index. thermal homeostasis: Chicks at 10 days of The present study was achieved to illustrate age were injected in wing vein with the role of desensitization of CAP-sensitive, hyperthermic (1 mg/kg, body weight) and TRPV1-independent pathway in ameliorating hypothermic (10 mg/kg, body weight) doses of the effect of thermal (cold/heat) and LPS from Escherichia coli (0111:B4; Sigma). inflammatory stimuli (LPS-induced fever/ LPS injected chicks showed signs of sickness hypothermia) through estimating heat loss behavior: lethargy, ruffled feathers and white index in chicken. diarrhea. For comparison, hypothermia and hyperthermia were induced by exposing chicks MATERIALS & METHODS: to an ambient temperature of 8°C and 38°C for 5 hours. The latter was considered a mild form 1-Experimental birds: Male, newly hatched of heat stress where panting behavior was not white leghorn chicks, specific pathogen free, observed in 10 days old chicken (Gerken et al., were brought from Goto Chick Company (Gifu, 2006). Japan) with a body weight range of 40±7 g. 4-Measurement of temperature: Colonic Chicks were kept on a 12:12 h light–dark cycle in thermostatically controlled cages. To match temperature (Tc) was measured by using a the chicks' requirements, the temperature of the lubricated thermistor probe as described cage for the newly hatched chicks was set at previously (Mahmoud et al., 2007). Skin 38°C and then decreased 0.5°C every day until temperature (Tsk) was measured by using day 4. All procedures were approved by the special kin probe (model XN-64, Technol Seven, Local Committee for Ethics of Animal Yokohama, Japan). To prevent stress fever, Experimentation, Care and Use of Gifu chicks were allowed to adapt to the handling University. and experimental cage (Jones et al., 1983), where the ambient temperature was kept at 2-Desensitization of capsaicin-sensitive 30°C. The thermistor probe was inserted gently

pathways: As described previously (Mahmoud 5 cm beyond the vent, and the colonic et al., 2007), CAP-sensitive pathways were temperature was monitored using a peripheral desensitized by repetitive injections of CAP into processor connected to a computerized medical newly hatched chicks. In brief, CAP (10 mg/kg; system (Chuo Electronic Co., Hong Kong). The Sigma, St. Louis, MO, U.S.A.) was dissolved in baseline temperature recordings were 0.1 ml of vehicle (10% ethanol, 10% Tween 80 determined for 1 h, and the chicks exhibiting no and 80% saline) and injected intravenously at 1, stress fever were used for experimentation. 2 and 3 days of age. Control groups were Each chick was used only once. To avoid injected with vehicle only.

-60- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

circadian variations in colonic temperature (SEM.) and values with asterisk means recordings, measurements were started at 8 a.m. significantly different (P<0.05).

Data in Table 1 showed the differences 5-Heat-loss index (HLI): The differences between colonic and surface temperatures (ΔT; between colonic and surface temperature (ΔT) Tc-Tsk) and Figures 1&2 illustrated ΔT and and heat loss index (HLI) were calculated from calculated sensible heat loss in terms of HLI at the following equation; ΔT=body temperature climax of body temperature. Chicks were (Tb)- Skin temperature (Tsk), and HLI = Tsk- exposed to mild heat (Ta 38°C) for 5 hours (Fig. (Ta)/(Tb)-(Ta). HLI value ranges theoretically 1) or injected intravenously with hyperthermic from zero (state of complete vasoconstriction) to doses of LPS (LPS 1 mg/kg, body weight Fig. 2). one (state of complete vasodilatation) (Steiner & Branco, 2002). In Fig. 1A chicks of control non treated groups, the ΔT ranged from 0.8 C to 1.0°C and 6-Statistical analysis: All statistical analysis the HLI at ambient temperature (Ta 25°C) was of data was performed using SPSS (2007) 0.95±0.02. Exposure to high Ta increased Software. All values were presented as surface temperature and lowered the ΔT during means±standard error (SEM). Descriptive the first 2 to 3 hr (Table 1: P<0.05), and statistics of data were analyzed by one way calculated HLI was 0.93 ±0.1°C at a 2.5 hr time analysis of variance (ANOVA) at time points of point (Fig. 1B). While heat exposure in CAP- climax and nadir of temperature curve. Tukey’s desensitized group elevated the surface HSD was used for comparisons among mean temperature, decreased the ΔT to a minimum of values. 0.2 ±0.1°C (Table 1: P<0.01), and increased HLI to 0.97±0.02 at a 2.5 hr time point (Fig. 1B). RESULTS & DISCUSSION: These results could be explained in part from the following facts; It is long known that TRPV1 1-Effect of CAP-desensitization on sensible heat loss after exposure to agonist (like CAP) affect vasomotor tone and hyperthermic stimuli in chicken at 10 causes hypothermia by skin vasodilatation days of age: (increased heat loss through the skin) and CAP (10 mg/kg, body weight, IV) was reduction in metabolism (a decreased oxygen injected at 1, 2 and 3 days of age. Chicks at 10 consumption; VO2 = decreased heat production, days of age were injected intravenously with Ayoub et al., 2009). However, this hyperthermic (1 mg/kg, body weight, IV) doses thermoteloerance to mild heat in CAP is of LPS. For comparison, hyperthermia was difficult to interpret at present study and needs induced by exposing chicks to an ambient further investigations. temperature of 38°C for 5 hours. Data are Although injection of a low dose of LPS presented as means and standard error bars (1 mg/kg, body weight, IV) which is known to

-61- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

cause fever (Mahmoud et al., 2007), ΔT was not Although injection of a low dose of LPS affected and HLI (0.94±0.02) was not different (1 mg/kg, IV) led to fever, ΔT was not affected from the control group (Table 1, Fig. 2 A &B). and HLI (0.94±0.02) was not different from the

Despite injection of LPS in CAP-desensitized control group (Table 1, Fig. 2). However, chicks (in absence of fever) did not change ΔT, injection of LPS in CAP-desensitized chicks HLI was increased to the maximum 0.99±0.01 at (absence of fever), did not change ΔT but, HLI 2.5 hr time point or climax of fever in LPS- was increased to the maximum 0.99±0.01 at 2.5 injected group (Fig. 2B). Increasing heat loss in hr time point (Fig. 2). Increasing heat loss in this experiment could explain in part our that experiment could explain in part our previous finding (Mahmoud et al., 2007) in previous study (Mahmoud et al., 2007) in which which CAP–pretreatment abolished LPS- CAP–pretreatment abolished LPS-induced induced fever in chicken through TRPV-1- fever in poultry chicken. independent pathway.

Table 1: Differences between colonic temperature and skin temperature (∆T) after exposure to hyperthermic stimuli in CAP-desensitized chicks at 10 days of age Time after Hyperthermic Stimuli Control Exposure [Heat (Ta 38°C) or LPS (1 mg/kg, IV)] (Ta; 25°C) (hour) Heat Heat /CAP LPS LPS/CAP -1.0 0.9±0.1 0.9±0.1 0.8±0.1 0.9±0.0 0.9±0.0

-0.5 1.0±0.0 0.8±0.1 0.9±0.1 0.9±0.1 0.9±0.1

0.0 0.9±0.1 0.9±0.0 0.9±0.0 0.8±0.0 0.8±0.0

0.5 0.8±0.0 1.0±0.1 1.0±0.1 0.9±0.1 0.9±0.1

1.0 0.9±0.1 1.9±0.0 0.8±0.1 1.0±0.1 1.0±0.0

1.5 0.9±0.0 0.8±0.0 0.6±0.1* 0.9±0.0 0.9±0.1

2.0 0.9±0.1 0.6±0.1* 0.4±0.0* 1.0±0.1 0.9±0.0

2.5 0.8±0.0 0.5±0.0* 0.2±0.1** 1.1±0.2 0.9±0.1

3.0 0.9±0.0 0.6±0.0* 0.4±0.0* 1.1±0.1 0.9±0.1

3.5 0.9±0.0 0.7±0.1 0.5±0.1* 0.9±0.0 1.0±0.0

4.0 0.8±0.1 0.8±0.0 0.6±0.1* 0.9±0.1 0.9±0.1

4.5 0.9±0.0 0.7±0.1 0.7±0.1 1.0±0.0 0.9±0.0

5.0 0.8±0.0 0.7±0.0 0.8±0.0 0.9±0.0 0.9±0.1

-62- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Fig.1 Fig.2 A A

1.6 1.6

C) C) C)

C) #

° ° ° ° 1.2 1.2

0.8

Tsk;Tsk; Tsk;

Tsk; 0.8

- - - - *

0.4 ** 0.4 **

T (Tc(Tc T T T (Tc T

T (Tc T 0.0

Δ Δ Δ Δ 0.0 Ta (25 °C) Ta (38 °C) Ta (38 °C) Control LPS LPS/CAP /CAP (1 mg, IV)

B B 0.95 0.93 0.97 0.96 0.94 0.99 1.0 1.0 0.9 0.9 0.8 0.8

0.7 (HLI)

0.7 (HLI)

(HLI)(HLI) 0.6 0.6 0.5 0.5 Ta (25 °C) Ta (38 °C) Ta (38 °C) Control LPS LPS/CAP (1 mg, IV) /CAP Fig. 1 & 2: Temperature differences (ΔT: Tc-Tsk) and heat loss index (HLI) 2.5 hr after exposure to heat (Fig. 1: Ta 38°C) or at a body temperature climax of LPS (Fig.2: LPS 1 mg/kg, body weight, IV)-induced fever in 10 days old chicken. Values with asterisk mean significant (P<0.05) differences versus # in Tukey’s HSD tests

2-Effect of CAP-desensitization on were shown in Table 2, and ΔT and HLI at sensible heat loss after exposure to nadir of body temperature were shown in hypothermic stimuli in chicken: Figures 3&4. CAP (10 mg/kg, IV) was injected at 1st, 2nd On other side, Data in Table 2 indicated and 3rd days of age. Chicks at 10 days of age that exposure to cold lowered skin temperature were injected intravenously with hypothermic and decreased the ΔT from time point of 1.5 hr (10 mg/kg, body weight) doses of on (P<0.05), and HLI reached the maximum lipopolysaccharide (LPS). For comparison, (0.99±0.01) as shown in Fig. 3. While cold hypothermia were induced by exposing chicks exposure in CAP-desensitized group did not to an ambient temperature of 8°C for 5 hours. affect the skin temperature and the ΔT started Data are presented as means and standard to decrease only after 4.5hr from cold exposure error bars (SEM.) and values with asterisk (Table 2). The controversial finding was that means significantly different (P<0.05) versus #. HLI in CAP-desensitized chicken was not Chicks at 10 days of age were exposed to increased but was similar to control non treated cold (Ta 8°C) for 5 hours or injected with group (0.94±0.02) at time point 2.5 hr (Fig. 3). hyporthermic doses of LPS (LPS 10 mg/kg, The present results indicated that the non body weight, IV). The differences between treated group responded immediately to cold colonic and surface temperatures (ΔT; Tc-Tsk) exposure by significantly increasing its HLI to

-63- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

the maximum level, however, CAP- and decreased the ΔT from time point of 1.5 hr desensitization clearly enhanced the chicks' on (P<0.05), and HLI reached the maximum ability to maintain on-chick body surface (0.99±0.01) as shown in Fig. 3. While cold temperatures during exposure to 8°C and can exposure in CAP-desensitized group did not recover much faster from cold exposure than affect the skin temperature and the ΔT started non-treated group (Nikami et al., 2008). to decrease only after 4.5hr from cold exposure Injection of high dose of LPS (10 mg/kg, IV) (Table 2). The controversial finding was that which is known to cause hypothermia in chicken HLI in CAP-desensitized chicken was not decreased ΔT along 1.0 to 3 hr time points post- increased but was similar to control non treated injection (P<0.05, Table 2). As a result HLI was group (0.94±0.02) at time point 2.5 hr (Fig. 3). increased to 0.97±0.01. But no such effect was The present results indicated that the non observed in CAP-desensitized group (Fig.4). treated group responded immediately to cold Preservation of heat loss by CAP-desensitization exposure by significantly increasing its HLI to after cold exposure could be explain in part the the maximum level, however, CAP- previous study (Nikami et al., 2008) in which desensitization clearly enhanced the chicks' CAP–pretreatment attenuated cold-induced ability to maintain on-chick body surface hypothermia in newly hatched chicken. temperatures during exposure to 8°C and can However ion case of LPS-induced hypothermia recover much faster from cold exposure than that could lead to shock, it is plaucible to relate non-treated group (Nikami et al., 2008). this action to the anti-oxidant effect of CAP as Injection of high dose of LPS (10 mg/kg, IV) reported by Nikami et al. (2008) rather than its which is known to cause hypothermia in chicken effect on vasomotor tone of skin indicated by decreased ΔT along 1.0 to 3 hr time points post- changes in heat loss index. injection (P<0.05, Table 2). As a result HLI was Values with asterisk mean significant increased to 0.97±0.01. But no such effect was (P<0.05) differences versus# in Tukey’s HSD observed in CAP-desensitized group (Fig.4). tests. Preservation of heat loss by CAP-desensitization Chicks at 10 days of age were exposed to after cold exposure could be explain in part the cold (Ta 8°C) for 5 hours or injected with previous study (Nikami et al., 2008) in which hyporthermic doses of LPS (LPS 10 mg/kg, CAP–pretreatment attenuated cold-induced body weight, IV). The differences between hypothermia in newly hatched chicken. colonic and surface temperatures (ΔT; Tc-Tsk) However ion case of LPS-induced hypothermia were shown in Table 2, and ΔT and HLI at that could lead to shock, it is plaucible to relate nadir of body temperature were shown in this action to the anti-oxidant effect of CAP as Figures 3&4. reported by Nikami et al. (2008) rather than its On other side, Data in Table 2 indicated effect on vasomotor tone of skin indicated by that exposure to cold lowered skin temperature changes in heat loss index.

-64- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 2: Differences between colonic temperature and skin temperature (∆T) after exposure to hypothermic stimuli in CAP-desensitized chicks at 10 days of age Time after Hyporthermic Stimuli Control Exposure [Cold (Ta 8°C) or LPS (10 mg/kg, IV)] (Ta; 25°C) (hour) Cold Cold/CAP LPS LPS/CAP -1.0 0.9±0.1 0.9±0.0 0.9±0.0 0.9±0.0 0.9±0.0 -0.5 1.0±0.0 0.9±0.1 0.9±0.1 0.9±0.0 0.9±0.0 0.0 0.9±0.1 0.9±0.0 0.9±0.0 0.9±0.0 0.9±0.0 0.5 0.8±0.0 0.6±0.0* 0.8±0.1 0.8±0.1 0.8±0.1 1.0 0.9±0.1 0.7±0.1 0.9±0.0 0.6±0.1* 0.6±0.0* 1.5 0.9±0.0 0.6±0.1* 0.9±0.1 0.5±0.0* 0.7±0.1 2.0 0.9±0.1 0.5±0.1* 1.0±0.1 0.4±0.1* 0.8±0.0 2.5 0.8±0.0 0.4±0.1* 1.2±0.0# 0.4±0.1* 0.9±0.0 3.0 0.9±0.0 0.5±0.1* 1.0±0.1 0.6±0.1* 0.9±0.1 3.5 0.9±0.0 0.6±0.1* 0.8±0.0 0.8±0.1 0.8±0.1 4.0 0.8±0.1 0.5±0.1* 0.7±0.1 0.9±0.0 0.9±0.1 4.5 0.9±0.0 0.6±0.1* 0.6±0.0* 0.8±0.1 0.8±0.0 5.0 0.8±0.0 0.5± 0.0* 0.6±0.1* 0.9±0.0 0.9±0.0

Fig.3 Fig.4

A A

1.6

C) C) °

1.6 ° C)

C) ** ° ° 1.2

1.2 *

Tsk;Tsk;

- -

Tsk; Tsk; 0.8 - - 0.8 # #

0.4 T T (Tc

0.4 T (Tc

T T T (Tc(Tc Δ

Δ 0.0

Δ Δ 0.0 Control LPS LPS/CAP Control Ta 8 °C Ta 8 °C/CAP (10 mg, IV)

B B 0.95 0.99 0.94 0.95 0.97 0.95 1.0 1.0 0.9 0.9

0.8 0.8

HLI HLI HLI

HLI 0.7 0.7

(HLI) (HLI) 0.6 0.6 0.5 0.5 Control Ta 8 °C Ta 8 °C/CAP Control LPS LPS/CAP (10 mg, IV) Fig. 3 & 4. Temperature differences (ΔT: Tc-Tsk) and heat loss index (HLI) 2.5 hr after exposure to cold (Fig. 3: Ta 8°C) or at a body temperature nadir induced by LPS (Fig. 4: LPS 10 mg/kg, body weight, IV) in 10 days old chicken

-65- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

To sum, a CAP-sensitive, a TRPV1- substrate of cold-seeking behavior in independent pathway is involved in thermal endotoxin shock. PLoS ONE 1 (1), e1. (heat and cold) and non-thermal stimuli (LPS- Ayoub, S.S., Hunter, J.C., and Simmons. D.L. induced fever and hypothermia), and CAP (2009): Answering the burning question pretreatment improved thermotolerance in of how transient receptor potential chicken. Since preservation of body vanilloid-1 channel antagonists cause temperature balance is crucial for survival of unwanted hyperthermia. Pharmacol Rev. newly hatching chicks, which are inevitably 61(3):225-7. exposed to a wide range of ambient temperature Chen, C.W., Lee, S.T., Wu, W.T., Fu, W.M., Ho, every day, It could be considered that the F.M., Lin, W.W. (2003): Signal present study has implications for the poultry transduction for inhibition of inducible industry. Chickens are indifferent to the nitric oxide synthase and cyclooxygenase- burning pain sensation induced by CAP (Mason 2 induction by capsaicin and related & Maruniak, 1983; Tewksbury and Nabhan, analogs in macrophages. Br. J. 2001), and we can simply add CAP to their feed. Pharmacol. 140:1077–87. :(In addition to the well known effect of hot chilli Gerken, M., and Afnan R., Dِrl, J. (2006 pepper (or CAP) on derratization of rodent Adaptive behaviour in chickens in (Tewksbury and Nabhan, 2001). relation to thermoregulation. Arch. Geflügelk., 70 (5). S. 199–207. Finally it could be concluded that, Jones, C.A., Edens, F.W., and Denbow, D.M. capsaicin desensitization enhanced (1983): Influence of age on the thermoregulatory tolerance to both thermal temperature response of chickens to stimuli in terms of cold and heat and to Escherichia coli and Salmonella inflammatory agent in part by affecting heat typhimurium endotoxins. Poultry Science loss mechanisms in newly hatched chicks. 62 (8), 1553–1558.

Kim, C.S., Kawada, T., Kim, B.S., Han, I.S., REFERENCES: Choe, S.Y., Kurata, T. (2003): Capsaicin exhibits anti-inflammatory property by Almeida, M.C., Steiner, A.A., Branco, L.G., and inhibiting IkB-degradation in LPS- Romanovsky, A.A. (2006a): Cold seeking stimulated peritoneal macrophages. Cell behavior as sa thermoregulatory strategy Signal 2003; 15:299–306. in systemic inflammation. The European Kluger, M.J. (1991): Role of pyrogens and Journal of Neuroscience 23 (12), 3359– cryogens. Physiological Rev. 71 (1), 93– 3367. 127. Almeida, M.C., Steiner, A.A., Branco, L.G., and Mahmoud, M.E., Shimizu, Y., Shiina, T., Romanovsky, A.A. (2006b): Neural Nikami, H., Dosoky, R.M., Ahmed, M.

-66- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

M., and Takewaki, T.(2007): Involvement Romanovsky, A.A. (2007): Thermoregulation: of a capsaicin-sensitive TRPV1- some concepts have changed. functional independent mechanism in architecture of the thermoregulatory lipopolysaccharide-induced fever in system. American Journal of Physiology, chickens. Comparative Biochemistry and Regulatory, Integrative and Comparative Physiology part A, Molecular and Physiology 292 (1), R37–R46. Integrative Physiology 148 (3), 578-583. Romanovsky, A.A., Shido, O., Sakurada, S., Mailman, D., Self, A., and Henry, M. (1999): Sugimoto, and N., Nagasaka, T. (1996): Time- and surgery-dependent effects of Endotoxin shock: thermoregulatory lipopolysaccharide on gut, cardiovascular mechanisms. American Journal of and nitric oxide functions. Shock 12 (3), Physiology, Regulatory, Integrative and 208–214. Comparative Physiology 270 (4 Pt 2), Mason, J.R. and Maruniak, J.A. (1983): R693–R703. Behavioral and physiological effects Romanovsky, A.A., Almeida, M.C., Aronoff, of capsaicin in redwinged blackbirds. D.M., Ivanov, A.l., konsman, J.P., Pharmacology Biochemistry and Steiner, A.A., and Turek, V.F., (2005): Behavior 19 (5), 857–862. Fever and hypothermia in systemic Nääs, I.A., Romanini, C.E.; Neves, D.P., inflammation: recent discoveries and Nascimento, G.R., and Vercellino, R.A., revisions. Frontiers in Bioscience 10, (2010): Broiler surface temperature 2193–2216. distribution of 42 day old chickens. Sci. Rudaya, A.Y., Steiner, A.A., Robbins, J.R., Agric. 67 (5): 497-502. Dragic, A.S., and Romanovsky, A.A. Nikami, H., Mahmoud, M.E., Shimizu, Y., (2005): Thermoregulatory responses to Shiina, T., Hirayama, H., Iwami, M, lipopolysaccharide in the mouse: Dosoky, R.M., Ahmed, M.M., and dependence on the dose and ambient Takewaki, T.(2008): Capsaicin temperature. American Journal of pretreatment attenuates LPS-induced Physiology, Regulatory, Integrative and hypothermia through TRPV1- Comparative Physiology 289 (5), R1244– independent mechanisms in chicken. Life R1252. Sciences 82: 1191–1195. Saia, R.S. and Carnio, E.C. (2006): Richard, S.A. (1971). The significance of Thermoregulatory role of inducible nitric changes in the temperature of the skin oxide synthase in lipopolysaccharide- and body core of the chicken in the induced hypothermia. Life Sci. 79: 1473– regulation of heat loss. J. Physiol. (1971), 1478. 216: 1-10. Shinder, D., Rusal, M., Tanny, J., Druyan, S. and Yahav, S. (2007): Thermoregulatory

-67- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

responses of chicks (Gallus domesticus) molecular bases of the initiation of fever. to low ambient temperatures at an early PLoS Biology 4 (9), e284. age. Poultry Science, 86 (10), 2200-2209. Steiner, A.A. and Branco, L.G. (2002): Hypoxia- SPSS (2007): Statistical package analysis of induced anapyrexia: implications and social sciences for window (1, 14 version). putative mediators. Annual Review of Steiner, A.A. and Ivanov, A.I., Serrats, J., Physiology 64, 263–288. Hosokawa, H., Phayre, A.N., Robbins, Tewksbury, J.J. and Nabhan, G.P. (2001): Seed J.R., Roberts, J.L., Kobayashi, S., dispersal Directed deterrence by Matsumura, K., Sawchenko, P.E., and capsaicin in chilies. Nature 412 (6845), Romanovsky, A.A. (2006): Cellular and 403–404.

ANNEX OF TERMS

CAP: Capsaicin: main ingredient of hot chilli pepper. LPS: Lipopolysachharide (outer membrane of Gram negative bacteria). HLI: Heat loss index. Tc: Colonic temperatures. Tb: Body temperature. Tsk: Skin temperatures. Ta: Ambient temperature. ∆ T: Differences between 2 temperatures or heat increment. TRPV1: Capsaicin receptors: transient membrane potential of vanilloid subtype-1 HLI: Heat loss index. Hypothermia: Lowered body temperature. Nadir: Lowest point at temperature curve in case of hypothermia. Hyperthermia: Elevated body temperature. Fever: Elevated body temperature in case of infection. Climax: Highest point at temperature curve in case of fever. ANOVA: One way analysis of variance.

-68- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

قياس الفقد الحراري المحسوس في كتاكيت الدجاج فاقد الحساسية للكابسايسيـن عند التعرض لالختالل في االتزان الحراري معتمد السيد محمود، ريم محمود دسوقي، مصطفى محمد أحمد، تاداشي تاكيواكي

الخلفية البحثية: الحفاظ على االتزان الحراري في كتاكيت الدجاا حجيثدة الف ده لدم ة قيدة ودن نقدن ة د نادا ا الرعايدة للحفاظ على كتاكيت الجاا حجيثة الف ه قن النفنق ن إحجاث التحقا الحراري عنج التعرض للادن البدارج ةن الحدار ةن ةثندا الحقى ةن الهبنط في جراة الحرارة القواحب للقرض.

الغرض قن الجرااة: ن ياه قعاقا الف ج الحراري في كتاكيت الجاا فا ج الحاااية للكابااياين عندج التعدرض للقداثرات الحرارية قثا: البرج نالحرارة نالعناقا القاببة للحقى ةن الهبنط في جراة الحرارة.

طدددرق البحدددث: تددد قعالادددة كتاكيدددت اللاهدددنرن امبددديض فدددى اليدددن امنا الثددداني الثالدددث بعدددج الف ددده بقددداجة الكاباايادددين )01 قج/كا قن النزن الحي( في النريج ينقيا نلقجة 3 ةيدا نللدإل دحدجاث ظدا رة ف دج الحااادية للكاباايادين نالتدي تد الكشف عنها عن طريق ف جان انخفاض جراة الحرارة الاا القواحب لح ن الكاباايادين نبعدج قدرنر ةادبنن قدن القعالادة تدد تعددرض قادداقيل قددن الكتاكيددت للقدداثرات التاليددة: البددرج )8° ق نيددة فددي رفددة التبريددج( الحددرارة )38° ق نيددة فددي ام فددا الحرارية( ح ن قركب الليبنبنلياكاريج بارعة )0 قج/كا قن النزن الحدي( فدي النريدج دحدجاث الحقدى كقدا تد ح دن ارعدة ةخر )01 قج/كا قن النزن الحي( في النريج دحجاث الهبنط في جراة الحرارة نفي اقيل له القاقنعات ت ياه جراة الحرارة الالج نال نلنن كا نوف ااعة نبجة ال يداه بدا الح دن باداعة نةيضدا لقدجة 5 اداعات بعدج الح دن نللدإل لحاداب قعجا الف ج الحراري القحانه عن طريق الالج.

النتا ج: قعجا الف ج الحراري كدان حدنالي 19.5 فدي كتاكيدت القاقنعدة الضدابطة الغيدر القعاقلدة نالتدي لد تتعدرض لهاهداج نفي جراة حرارة الغرفة ةقا في الكتاكيت التي ت قعالاتها بالكابااياين ت زياجة الف ج الحراري عنج تعرضدها لكدا قدن الحدرارة العاليددة )38° ق نيددة فددي ام فددا الحراريددة( ةن بعددج ح ددن ارعددة الليبنبنلياددكاريج القاددببة للحقددى علددى الن دديض فددإن تدديثير الكابااياين ل ي تور ف ط على زياجة الف ج الحراري نلكن ةيضا ةج إلى الت ليدا قدن الف دج الحدراري عندج التعدرض للبدرج )°8 ق نية في رفة التبريج(.

االاتنتا : يعتبر التيثير القعالج للكاباياين فى التنظدي الحد ارري للكتاكيدت تديثي ار قزجنادا ةثندا التعدرض للادن الحدار لحقدى الليبنبنلياددكاريج ةن ةثنددا تعددرض الكتاكيددت للبددرج. ن ددج يعددل للددإل لقددج كفددا ة الكاباياددين فددى الزيدداجة ةن الت ليددا قددن الف ددج الحد ارري ادنيا . نلهدله الجرااددة ة قيدة تطبي يددة خاوددة فدى حقايددة الكتاكيدت الوددغيرة عنددج تعرضدها لقدداثرات التغيدر فددى جراددات الحرارة القحيطة للكتاكيت نالتي تاجي الى اختالا في التنطي الحراري لوغار الكتاكيت نقن ث النفنق.

-69- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

STATUS OF SOME NUTRIENTS AND HEAVY METALS WITH RELATIONS TO THEIR BEARING MINERALS IN THE SOILS OF NILE TERRACES AT ASSIUT, EGYPT Mohamed El-Ameen Abd El-kareem Faragallah Soil and Water Sci. Dept., Faculty of Agric., Al-Azhar University, Assiut, Egypt

ABSTRACT:

Nine soil profiles were selected from various Nile terraces located south Assiut barrage at Assiut city, Egypt to a) examine levels and distributions of some available nutrients (P, K, Fe, Mn, Zn and Cu) and some heavy metals (Ni, Pb and Cd), b) clear up their relations with some physical and chemical soil properties, and c) evaluate their relations to their bearing minerals in the soils of the most recent and old Nile terraces. Results showed soil available phosphorus values are higher in the oldest terraces on both Nile sides than in the youngest ones. However, most soil layers of the youngest Nile terraces show greater available potassium levels than the oldest ones. The recent and old Nile terraces soils exhibit very high level of P and medium amounts of K. In most cases, there are no consistent changes in the levels of either available phosphorus or potassium with depth. Highly significant positive correlations were obtained between the available P and K and soil contents of silt, clay, organic matter and CEC. On the other hand, highly significant negative

correlations were recorded with sand, pH and CaCO3 contents. Significant positive correlations were found between the phosphorus levels and both monazite and the sum of apatite + monazite minerals. However, insignificant, positive correlations between available potassium and its bearing minerals were recorded, except the case of feldspars in the clay fraction was a highly significant, positive correlation. The highest levels of DTPA-extractable Fe are obtained in the oldest Nile terrace soils. Whereas, extractable Mn levels are high in the youngest terraces on both Nile sides. Moreover, the soils of the youngest and oldest terraces are Fe and Mn sufficient. However, most soils of the terrace bench in the interference zone and the terrace rear suture in desert fringes are considered Fe and Mn deficient. The levels of soil extractable Cu are quite variable among the youngest and oldest Nile terraces, and all the investigated soils are Cu-nondeficient. On the other hand, most examined soils are considered Zn- deficient. No clear pattern of distributions of these investigated micronutrients was observed with depth. Highly significant positive correlations expressed the relations between fine particles, organic matter and CEC contents and extractable Fe, Mn and Cu. However, highly significant negative

correlations were observed with amounts of sand, CaCO3 and soil pH. The DTPA-extractable Zn did not show any significant correlations with all the considered soil properties. The extractable Fe and Cu

-69- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

showed highly significant positive correlations with epidotes, biotite and feldspars of both sand and clay fractions. Also, the extractable Mn has significant positive correlations with epidotes, hornblende, smectites and chlorite. However, low positive correlations occurred between the extractable Zn with both hornblende and pyroxene. Generally, DTPA-extractable Ni and Pb are slightly higher in the youngest Nile terrace soils than in the oldest ones, without any clear trend of their distributions with depth. Moreover, the youngest and oldest Nile terraces soils contain similar vey low amounts of extractable Cd. Correlations between extractable Ni and contents of clay, silt, organic matter and CEC are highly significant and

positive, whereas, they are highly significant and negative with sand, pH, ECe and CaCO3 contents. The extractable Pb has significant positive correlations with the soil contents of silt, clay, and CEC, but,

negative significant correlations with both sand and CaCO3 contents. Slightly significant positive correlations between the extractable Ni and Pb and hornblende are recorded. Both biotite and chlorite were highly significant positive correlated to Ni.

INTRODUCTION: Two main sources of heavy metals in the soil include: (i) the natural background, which Sediment transfer from continents to oceans represents the heavy metal concentration that is or seas via rivers is one of the important derived from parent rocks, and (ii) the processes regulating river-bank stabilization, anthropogenic contamination, that represents soil formation, biogeochemical cycling of the human applications of agrochemicals, elements and many other earth-related organic amendments, animal manure, mineral processes (Chakrapani, 2005). Aswan High Dam fertilizers, industrial manufacturing wastes, construction and increased human activities in domestic refuse and waste materials. Generally, the Nile valley have drastically changed the there are more heavy metals in soils originated chemistry and mineralogy of the river flux. from anthropogenic sources than natural Dissolved transport is a major process for Ca, sources (Nriagu and Pacyna, 1988; Stehouwer K, Sr, Zn, Cu, Ni and V. Manganese, Fe and Cr et al., 2000; Jjemba, 2004; Wei et al., 2007; are mainly carried by suspended matter. Along Yadav et al., 2009). Iron, Zn, Cu and Ni in the course of the river, the distribution of alluvial deposits are mainly originated from the elements in the suspended matter and sediments terrestrial (rock and soil) weathering that is is generally controlled by natural processes, the considered a natural source. However, Pb, Ni, relative importance of elemental transport Zn, Cu and Cd may come from some phases, as well as the oxidation, precipitation anthropogenic sources (Jakovljevic et al., 1997; and sedimentation of mineral species through Zhou et al., 2004; Jjemba, 2005; Jinling et al., the varying physico-chemical conditions of the 2009). environment. Pollution input in the Nile There is a great diversity of possible particulate load is not major, compared to the associations of trace elements with the various natural ones (Dekov et al., 1997). substrates. These elements can be adsorbed on

-70- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

the particle surface of clays or organic matter physical and chemical soils properties, and and present in the lattice of secondary and (3) evaluate their relations to bearing-minerals primary minerals (Tessier and Campbell, 1988). in the soils of the most recent and old Nile The grain size and heavy minerals control the terraces at Assiut, Egypt. distribution of chemical elements in alluvial soils (Zhou, et al., 2004). The fine sand fraction MATERIALS AND METHODS: in soils is likely to have been derived from the Nine profiles were selected to cover various more easily weathered minerals, which are also Nile terraces located south Assiut barrage, the main source of trace elements (Sillanpaa, Assiut governorate. Profiles 1 and 5 represented 1972). The bioavailability of metals in soils is a the most recent Nile terraces in the eastern and dynamic process that depends upon specific western banks of the Nile stream, respectively. combinations of chemical, biological, and Profiles 2, 6 and 7 represented the succeeding environmental parameters (Li and Thornton, terraces; profiles 2 and 7 are scarps of flood 2001; Peijnenburg and Jager, 2003; Panuccio et plain in the eastern and western sides of the Nile al., 2009). Mohamed (1990) found that soil River, respectively. Profiles 3 and 8 represented texture, mineral composition, CaCO3, organic the terrace bench or plain that located in the matter and pH were the most important factors Nile valley-desert interference zone which they that affect the total and available micronutrients are close to the eastern and western desert, in both alluvial and calcareous soils. respectively. Profiles 4 and 9 represented the Several investigators reported the levels of terrace rear suture that presented in the fringes various extractable nutrients and their of the eastern and western desert, respectively correlations with some properties of alluvial soils (Fig. 1). (Khalil et al., 1990; Amer et al., 1991; El-Shazely Soil samples were taken from the layers of et al., 1991; Farragallah, 1995; Khalifa et al., the profiles that represent the studied Nile 1996; El-Desoky, and Ghallab, 2000; Ghallab, terraces, air-dried, crushed, sieved with a 2 mm 2002; Abdel-Mawgoud and Faragallah, 2004; sieve and kept for physical, chemical and Faragallah and Essa, 2005; Gomah et al., 2007). mineralogical analysis. The particle-size Few researches have been focused on the relations distribution of the soil samples was performed between various extractable nutrients and heavy according to Piper (1950) and Jackson (1973). metals and their bearing-minerals in alluvial Organic matter in the soil samples was soils. determined using Walkely-Black method The current work was carried out to (Jackson 1973). Soil calcium carbonate was (1) examine levels and distributions of available measured by the calcimeter method, according some macro- micronutrients (P, K, Fe, Mn, Cu to Nelson (1982). Soil pH was measured in a 1:1 and Zn) and some heavy metals (Ni, Pb and Cd), water suspension of soil to water using a glass (2) clear up their correlations with some

-71- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

electrode as reported by Mclean (1982). The according to the method described by Lindsay electrical conductivity (ECe) was measured in and Norvell (1978) and determined using atomic the saturated soil paste extract using a absorption (model Perkin-Elmer 1999). conductivity meter. Soluble ions were also Heavy and light minerals were separated determined in the saturated soil paste extract from fine and very fine sand fractions (0.25 to using the methods of Jackson (1973). Cation 0.063 mm) using the procedure of Brewer exchange capacity (CEC) of the soil samples was (1964). These minerals were identified determined using NaOAC at pH 8.2 as a according to Milner (1962). Percentages of the saturating solution and NH OAC at pH 7.0 as a 4 heavy and light minerals in the fine and very displacing solution, and then sodium was fine sand fractions of the studied soil samples measured by flamephotometer (Jackson, 1973). are given in Table (2). These examined physical and chemical The clay-size fractions were separated from properties of the studied soil samples are the studied soil samples, prepared and present in Table (1). investigated using X-ray diffractometer. The Available P and K were extracted and clay minerals were identified and their relative determined according to Soltanpour and proportions were determined according to Schwab (1977). Available iron (Fe), zinc (Zn), Schultz (1964). Relative abundance of the manganese (Mn), copper (Cu), nickel (Ni), lead identified minerals in the clay fraction of the (Pb) and cadmium (Cd) were DTPA-extracted studied soil samples is present in Table (3).

-72- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Assiut Barrage 4

A ssiut 3 2

City 1

5 6 7 Mediterranean Sea

Cairo 8

9 Assiut Red Sea

0 1 2 3 Km Aswan Profile Location Fig. 1: Location map of the studied Nile terraces south Assiut barrage

-70- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

-73- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

الجدول )2(

-74- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

الجدول )3(

-75- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

RESULTS AND DISCUSSIONS: the organic residues are possibly the cause for the very high available P values obtained. Phospho- I-Available nutrients: rus-bearing minerals in desert could be the source 1-Phosphorus: for the obtained P values. These results are in The available phosphorus in the soil samples agreement with those of Farragallah (1995) and of the studied profiles is present in Table (4). The Abdel-Mawgoud and Faragallah (2004). results indicated that the available phosphorus The relationships between soil available varies from 14.84 to 54.68 ppm. Most layers of phosphorus and particle size distribution as well the soils of recent and old Nile terraces show as some chemical properties have been greater P concentrations than those of the terrace statistically studied as correlation coefficients

bench in the interference zone and those of the (r values) (Table 5). The highest positive terrace rear suture in desert fringes on both Nile correlation coefficient (r=+0.755**) was recorded sides, indicating that Nile sediment-derived soils between the sum of silt+clay fractions and are characterized by high contents of available phosphorus. However, the highest phosphorus. In general, the soils of the oldest negative correlation coefficient (r=-0.755**) terraces on both Nile sides have higher values of occurred with sand fraction. Such results are in available phosphorus than those of the youngest accordance with those of Amer et al. (1991), Eden ones, particularly in the surface layers. This could and Parfitt (1992), Awadalla (1993) and be due to the surface applications of organic and Faragallah (1995). inorganic phosphate fertilizers during cultivating Highly significant positive correlations were the soils of the oldest terraces on both Nile sides. recorded between each of clay, silt, organic In most cases, there are no consistent changes matter and CEC and available phosphorus. in the available phosphorus in the studied soils However, highly significant negative correlations with depth as shown in Figure (2). This reflects were obtained between soil CaCO3 as well as the nature of sediments forming these soil profiles pH and available P (Table 5). Insignificant and their role in affecting its chemical correlation was observed between salinity content composition. (ECe) of the studied soils and available P with All the studied soils of the recent and old Nile very low negative correlation (r=-0.130). Most of terraces show a very high level of phosphorus these results agree with those reported by (>21ppm), according to Cook (1967). However, Awadalla (1993) and Faragallah (1995). most soil layers of the terrace bench in the Concerning, the relationship between interference zone and of the terrace rear suture in available phosphorus and its bearing-minerals in desert fringes on both Nile sides exhibit moderate the studied soils, correlation coefficients between to high values of available P (13-20 ppm). For the the concentration of available P and the oldest Nile terrace soils, the sediment nature as percentages of apatite, monazite and apatite + well as the application of mineral fertilizers and monazite were calculated and are given in Table

-76- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

(6). The results showed that significantly positive suture in desert fringes on both Nile sides have correlations are between the available phosphorus relatively low values of available K (48.71 - 186.50 and both monazite (r=+0.319*) and the sum of ppm). No specific pattern of potassium apatite+monazite (r=+0.350*). However, low distribution with depth is evident (Figure 3). The positive correlation is found between available P nature of sediments and the fertilizer applications and the percentages of apatite (r=+0.280). These could be the reason for enriching the surface soil data suggest that monazite could be the possible with K+. Similar results were obtained by El- source of available P in the uncultivated soils of Hadedy (1976), Faragallah (1995) and Abdel- the youngest Nile terraces on river border and the Mawgoud and Faragallah (2004). terrace rear suture in the desert fringes. The correlation coefficients between available Moreover, the phosphorus bearing minerals as potassium and some physical and chemical well as the organic and inorganic fertilizers added properties of the soils under study are given in through cultivation practices could be the main Table (5). Highest positive correlation coefficients source of available P in the cultivated soils of the were recorded between clay and organic matter oldest Nile terraces. contents (r=+0.643** and +0.673**, respectively) and the available potassium concentration. On the 2-Potassium: other hand, the available K showed a highest The available potassium in the studied soil negative correlation (r=-0.541**) with the sand profiles is shown in Table (4) and graphically fraction. presented in Figure (3). The results generally Highly significant, positive correlations were indicated clear variations in the available K obtained between the available K and contents of among various layers and sites. Most soil layers of silt, clay+silt and CEC (r=+0.400**, +0.565** and the youngest Nile terraces show a greater K +0.421**, respectively). However, highly concentration than the oldest ones, but the surface significant, negative correlations were recorded layers of the oldest terraces have the highest between the available K and both soil pH and amounts of available potassium. According to CaCO3 content (r=-0.599** and -0.510**, Metson (1961), most layers of youngest terrace respectively). Insignificant negative correlation soils show medium amounts of K (205.04 - 293.50 coefficient was found between available K and soil ppm) on both Nile sides. However, most layers of salinity (Table 5). The same trends reported by oldest terrace soils exhibit low to medium Abou-El-Roos (1972), Kabata-Pendias and amounts of K (155.68 -256.50 ppm). The surface Pendias (1984) and Faghami et al. (1985). layer of the oldest terraces soils on eastern and Regarding the relation between the available western Nile sides show very high amounts of K potassium and its bearing minerals, correlation (571.20 and 474.51 ppm, respectively). On the coefficients between levels of available K and the other hand, most soil layers of the terrace bench percentages of microcline, feldspars and biotite in in the interference zone and the terrace rear sand fraction as well as feldspars, smectites, and

-77- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

micas in clay fraction were calculated and present clearly expressed. The relatively high K levels in

in Table (6). A highly significant, positive the cultivated oldest Nile terrace soils could be correlation was only obtained between the attributed to agricultural practices rather than available K and the k-feldspar content of the due to K-bearing minerals. The uncultivated soils clay fraction (r=+0.540**). In all cases, of youngest Nile terraces on river border and the correlation values are positive and insignificant. terrace rear suture in the desert fringes have Accordingly, the role of microcline and biotite relatively high values of available K due to the (sand fraction) as well as smectites, and micas total native amounts that have not been yet used (clay fraction) in affecting available K is not by cropping.

Table 4: Available phosphorus (P) and potassium (K) in soil samples of the studied Nile terraces Profile Depth P K Profile Depth P K No. (cm) (ppm) No. (cm) (ppm) 0 - 15 43.22 227.70 0 - 20 48.92 281.15 15 - 25 48.29 256.50 5 20 - 35 43.6 272.92 1 25 - 35 39.20 293.50 35 - 65 40.99 205.04 35 - 70 49.97 412.80 0 - 45 38.25 192.70 6 0 - 40 54.68 571.20 45 - 80 42.57 180.36 40 - 70 37.71 198.90 0 - 40 54.43 474.50 2 70 - 95 28.04 256.50 40 - 70 51.32 155.68 7 95 - 195 26.30 190.60 70 - 120 40.24 69.28 195- 235 48.79 221.50 120- 155 32.65 205.04 0 –35 38.08 366.75 0 –50 25.18 295.55 35 –55 39.83 166.00 50 –70 18.70 75.45 3 8 55 –70 20.95 131.00 70 –85 13.84 56.94 70 –150 20.20 178.30 85 –150 20.93 48.71 0 - 20 35.24 65.17 0 - 10 36.63 279.41 4 20 - 35 36.94 186.50 9 10 - 35 20.04 114.54 35 - 150 20.93 71.34 35 - 150 20.54 126.88

Fig. 2: Distribution of available phosphorus in the soil profiles with distance from the Nile border to the desert on both sides

-78- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 5: Correlation coefficients (r values) of available P and K and some soil properties r value Soil property Phosphorus Potassium Sand (%) -0.755** -0.565** Silt (%) +0.655** +0.400** Clay (%) +0.689** +0.643** Silt + Clay (%) +0.755** +0.565** Organic matter (%) +0.555** +0.673**

CaCO3 (%) -0.740** -0.510** pH (1:1) -0.492** -0.599** -1 ECe (dsm ) -0.130 -0.157 CEC (Cmol+/kg) +0.545** +0.436**

Table 6: Correlation coefficients of available phosphorus and potassium and their bearing minerals of the studied soils Nutrient (ppm) Mineral (%) "r" value Apatite +0.280 Phosphorus Monazite +0.319* Apatite + Monazite +0.350* Microcline +0.278 Biotite +0.255 Potassium K-feldspars (clay fraction) +0.540** Smectites +0.281 Micas +0.160 * Significant at the 5% level. ** Significant at the 1% level.

Fig. 3: Distribution of available potassium in the soil profiles with distance from the Nile border to the desert on both sides

-79- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

3-Iron: positive correlations showed the relations between The DTPA-extractable Fe of the soil samples the organic matter content and CEC of the of the studied Nile terraces are present in Table studied soil samples and the DTPA-extractable (7) and graphically illustrated in Figure (4). Fe (r=+0.691** and +0.784**, respectively). Generally, it ranges between 1.33 and 36.72 ppm. However, highly significant negative correlations In soil samples representing the youngest and between both calcium carbonate content and soil oldest Nile terraces, the extractable Fe ranges pH and the DTPA-extractable Fe were obtained from 19.91 to 36.72 ppm. The highest levels of (r=-0.858** and -0.581**, respectively). A DTPA-extractable Fe are obtained in the oldest significant negative correlation was recorded Nile terraces soils. According to the limits of Viets between the soil ECe and the DTPA-extractable and Lindsay (1973), all samples of the youngest Fe (Table 8). Such trends were also found by and oldest Nile terraces soils and most soil layers Rabie et al. (1989), Amer et al. (1991), Awadalla of the terrace bench in the interference zone (1993) and Faragallah (1995). contain adequate amounts of available Fe (>4.5 Respecting the relation between the DTPA- ppm). However, most soil samples of the terrace extractable iron and its bearing minerals rear suture in desert fringes on both Nile sides are (opaques, epidotes, amphiboles, pyroxenes, deficient (<2.5 ppm) to marginal (2.5-4.5 ppm) biotite and feldspars according to Wedepohl, with respect to the extractable Fe. Similar results 1974 and Dress and Wilding, 1978, as well as were reported by Abdellah (1984), Amer et al. smectites, chlorite, sepiolite, palygorskite, micas (1991), Faragallah (1995), Abdel-Mawgoud and and vermiculite according to Duane and Robert, Faragallah (2004) and Faragallah and Essa 1989 and Velde, 1992), the correlation (2005). coefficients between them were calculated and No clear distribution pattern of the available listed in Table (9). The data reveal that highly Fe can be observed in all studied sites, especially significant positive correlations between the in the case of youngest and oldest Nile terraces extractable Fe and biotite (r=+0.817**) and soils (Figure 4). feldspars of the sand fraction (r=+0.615**) as The correlation coefficients (r values) well as feldspars of the clay fraction and between the DTPA-extractable Fe and some soil smectites (r=+0.749** & +0.425**, respectively). properties of the studied samples are given in A significantly positive correlation is found Table (8). The fine particles (clay and the sum of between the extractable Fe and both epidotes clay+ silt) showed the highest positive correlations (r=+0.335*) and chlorite (+0.375*), but low with the extractable Fe (r=+0.811** and positive correlations were recorded with +0.833**, respectively). On the other hand, the amphiboles (r=+0.149), micas (+0.292) and sand fraction was negatively correlated to the vermiculite (+0.167), indicating the role of these extractable Fe (r=-0.833**). Highly significant, minerals as a source for iron in the studied soils.

-80- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 7: DTPA-extractable Fe, Mn, Cu and Zn in the soil samples of the studied Nile terraces Profile Depth Fe Mn Cu Zn Profile Depth Fe Mn Cu Zn No. (cm) (ppm) No. (cm) (ppm) 0 - 15 23.79 7.67 2.41 0.4 0 - 20 24.93 8.78 2.45 0.79 15 - 25 21.71 8.54 2.39 0.39 5 20 - 35 25.79 8.57 2.89 1.43 1 25 - 35 28.62 3.43 1.58 0.31 35 - 65 26.04 7.08 2.77 0.38 35 - 70 23.08 4.16 2.56 0.45 6 0 - 45 31.10 4.46 2.84 1.69 0 - 40 19.91 4.73 1.96 0.66 45 - 80 29.81 6.44 2.45 0.52 40 - 70 36.72 3.56 2.07 0.42 0 - 40 20.52 1.61 2.08 0.31 2 70 - 95 29.33 3.61 3.20 0.4 7 40 - 70 26.67 1.74 2.10 0.38 95 - 195 32.70 2.69 2.96 0.37 70 - 120 26.79 1.08 2.40 0.32 195-235 33.26 2.23 3.44 0.58 120-155 35.32 1.51 2.67 0.67 0 - 35 13.37 5.84 1.44 1.19 0 - 50 12.41 4.25 1.88 0.46 3 35 - 55 8.24 1.26 0.82 0.75 8 50 - 70 6.30 0.52 0.68 0.34 55 - 70 8.41 1.16 0.93 1.20 70 - 85 3.76 1.93 0.41 0.18 70 - 150 6.37 0.37 0.41 0.22 85 - 150 4.24 2.00 0.71 0.39 0 - 20 1.80 0.17 0.23 0.16 0 - 10 20.46 4.78 1.39 1.52 4 20 - 35 9.01 0.97 0.72 0.47 9 10 - 35 3.68 3.12 0.21 0.19 35 - 150 1.33 4.82 0.27 0.56 35 - 150 1.48 0.31 0.20 0.12

Fig. 4: Distribution of DTPA-extractable Fe, Mn, Cu and Zn in the soil profiles with distance from the Nile border to the desert on both sides

-81- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 8: Correlation coefficients (r values) of DTPA-extractable Fe, Mn, Cu and Zn and some soil properties r value Soil property Fe Mn Cu Zn Sand (%) -0.833** -0.565** -0.824** -0.289 Silt (%) +0.688** +0.512** +0.636** +0.251 Clay (%) +0.811** +0.486** +0.863** +0.264 Silt + Clay (%) +0.833** +0.565** +0.824** +0.289 Organic matter (%) +0.691** +0.392* +0.743** +0.164 ** ** CaCO3 (%) -0.858** -0.463 -0.877 -0.176 pH (1:1) -0.581** -0.522** -0.681** -0.057 -1 ** ECe (dsm ) -0.338* -0.190 -0.400 -0.059 CEC (Cmol+/kg) +0.784** +0.424** +0.830** +0.318*

Table 9: Correlation coefficients of DTPA-extractable Fe, Mn, Cu and Zn and their bearing-minerals in the studied soils Nutrient Mineral R Value Nutrient Mineral R Value Opaques -0.453** Chlorite +0.339* Epidotes +0.335* Smectites +0.357* Mn Amphiboles +0.149 Sepiolite -0.103

Pyroxenes -0.256 Palygorskite -0.103 Biotite +0.817** Micas -0.034 Fe Feldspars (Sand Fraction) +0.615** Opaques -0.477** Feldspars (Clay Fraction) +0.749** Epidotes +0.497** Chlorite +0.375* Hornblende +0.224 Smectites +0.425** Augite -0.077 Sepiolite -0.211 Biotite +0.766** Cu Palygorskite -0.401** Garnet -0.517** Micas +0.292 Rutile -0.469**

Vermiculite +0.167 Sphene +0.163 Opaques -0.430** Feldspars (Sand Fraction) +0.574** Epidotes +0.425** Feldspars (Clay Fraction) +0.736** Hornblende +0.315* Opaques -0.287 Mn Garnet -0.362* Hornblende +0.190 Zn Rutile -0.394** Pyroxenes +0.233 Sphene -0.058 Biotite +0.073 * Significant at the 5% level. ** Significant at the 1% level.

4-Manganese: suture in desert fringes on both sides have amounts of DTPA-extractable Mn less than 1.0 Generally, the DTPA-extractable Mn of the ppm. Therefore, these soils are considered Mn studied soils ranges between 0.17 and 8.78 ppm deficient. No specific pattern of extractable Mn (Table 7). For the soil samples of the youngest and distribution with depth can be shown (Figure 4). oldest terraces, it varies between 1.08 and 8.78 These results are in full accordance with those ppm. According to DTPA-extractable Mn limits reported by Abd El-Razek et al. (1984) and of Viets and Lindsay (1973), all these soils are Faragallah (1995), Abdel-Mawgoud and Mn-sufficient. Moreover, the soils of the youngest Faragallah (2004) and Faragallah and Essa terraces on both Nile sides contain higher DTPA- (2005). extractable Mn than those of the oldest ones. The correlation coefficients calculated to test However, several soil samples of the terrace bench the effect of some soil properties on the level of in the interference zone and the terrace rear

-82- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

DTPA-extractable Mn, showed highly significant where rutile, opaques and garnet are considered positive correlations with contents of silt, clay, the most stable minerals. silt+clay and CEC (Table 8). On the other hand, highly significant negative correlations were 5-Copper: found in cases of sand, CaCO3 and pH of the Table (6) reveals that the DTPA-extractable studied soils. A significant positive correlation Cu of the investigated soils falls in the range (r=+0.392*) was obtained between the soil between 0.20 and 3.44 ppm. It can be stated that organic matter content and the extractable Mn. all these soils are considered Cu nondeficient Insignificant correlation resulted between soil according to the limits of Viets and Lindsay

ECe and DTPA-extractable Mn (r=-0.190). (1973), where they contain more than 0.2ppm. Similar trends were reported by Attia (1988), The youngest and oldest Nile terraces soil samples Rabie et al. (1989), El-Shazely et al. (1991) and have high levels of extractable Cu., Moreover, Amer et al. (1991). these levels were quite variable among these Manganese is present in magnetite terraces. This suggests that metal content may be (opaques), hornblende, epidote, garnet, rutile controlled by similar soil components. From the and sphene (Gladilovich and Sergeeva, 1971) as results in Table (7) and Figure (4), there is no well as smectites, chlorite, sepiolite, palygorskite clear trend in distribution of DTPA-extractable and micas (Duane and Robert, 1989 and Velde, Cu with depth of the studied soils. 1992). To clear up the relationship between the Highly significant correlations were found DTPA-extractable Mn and its bearing minerals, between the examined soil properties and the the correlation coefficients are recorded in Table DTPA-extractable Cu (Table 8). The highest ** (9). The obtained data clearly show that there are positive correlation coefficient (+0.863 ) was a highly significant positive correlation value obtained between the clay fraction and the between extractable Mn with epidotes DTPA-extractable Cu. On the other hand, the ** (r=+0.425**) and slightly significant positive highest negative correlation coefficient (-0.877 ) correlations with hornblende (r=+0.315*), was recorded with the CaCO3 content. These smectites (r=+0.357*) and chlorite (r=+0.339*). results well coincide with those obtained by Attia These suggest that epidote, hornblende, (1988), Rabie et al. (1989), Amer et al. (1991), smectites and chlorite minerals are considered a Faragallah (1995) and Marto-Anez and Motto source of manganese in the studied soils. (2000). However, there are highly significant negative With regard to the relation between the correlation values between the extractable Mn DTPA-extractable Cu and its bearing minerals, and both opaques (r=-0.430**) and rutile correlation coefficients were calculated between (r=-0.394**). Moreover, a slightly significant them, where copper occurs in hornblende, negative correlation for the extractable Mn with augite, biotite, feldspars, opaques (ilmenite & garnet (r=-0.362*). These relations are logic magnetite), epidotes, garnet, rutile and sphene

-83- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

(Mitchell, 1964; Gladilovich and Sergeeva, 1971). the terrace bench in the interference zone and the Data in Table (9) indicate that highly significant terrace rear suture in the desert fringes on both positive correlations were obtained between the sides, most samples are considered Zn deficient. extractable Cu and biotite (r=+0.766**), No regular distribution pattern is noticed for the feldspars of the clay fraction (r=+0.736**), the extractable Zn throughout the studied profiles feldspars of sand fraction (r=+0.574**) and (Table 7 and Figure 4). epidotes (r=+0.497**). Moreover, low positive The results in Table (8) indicate that no correlation with hornblende (r=+0.224). On the significant correlations occurred between all the other hand, highly significant negative considered soil properties and the DTPA- extractable Zn, except of soil CEC (r=+0.318*). correlations were recorded between the Fine soil particle i.e., silt, clay and clay+silt extractable Cu and garnet (r=-0.517**), rutile showed low positive correlations with the (r=-0.469**) and opaques (r=-0.477**). The extractable Zn (r=+0.251, +0.264 and +0.289, minerals of biotite, feldspars, hornblende and respectively). However, a low negative correlation epidotes are considered as a source of Cu in the (r=-0.289) was recoded with the sand fraction. investigated soils. Theses results are reliable Very low negative correlations were found with since these minerals are unstable but the others both soil pH and ECe (r=-0.057 and -0.059, i.e., rutile and garnet are ultrastable. respectively). Similar relations were obtained by

Khetawat and Vashishtha (1977), Ghoneim et al. 6-Zinc: (1984), Attia (1988) and Marto-Anez and Motto The DTPA-extractable Zn in the studied soils (2000). ranges from 0.12 to 1.69 ppm (Table 7). Soil Zinc is found mainly in pyroxenes, biotite, samples representing the youngest Nile terraces magnetite (opaques) and hornblende minerals show variable zinc concentrations, ranging from (White, 1957 and Wiklander, 1958). Based on 0.31 to 1.69 ppm. According to the limits of Viets such information, correlation coefficients were and Lindsay (1973), most of these soils are calculated between these minerals and the DTPA- considered Zn deficient since they contain less extractable Zn of the studied soils (Table 9). The than 0.5 ppm. Two samples (the surface sample of obtained data clearly illustrate that there are low profiles 5 and the subsurface of profile 6 are Zn positive correlations between the extractable Zn marginal. The surface layer of profile 6 and the and hornblende (r=+0.190) and pyroxenes subsurface one of profile 5 are only Zn (r=+0.233). Moreover, a very low positive nondeficient since they have more than 1.0 ppm. correlation with biotite (r=+0.073). The only The DTPA-extractable Zn of the oldest Nile exception appears with opaques, which show a terraces soils falls between 0.31 and 0.67 ppm. low negative correlation (r=-0.287). These data Only, three samples (the surface and lower layers indicate that the role played by these suggested of profile 2 and the lower one of profile 7) are Zn minerals in affecting the level of available Zn is marginal but the rest are Zn deficient. For soils of not clearly expressed.

-84- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

II-Heavy metals: On the other hand, the highest negative correlation coefficient (r=-0.801**) was calculated 1-Nickel: for the CaCO3 content, followed by the sand The DTPA-extractable Ni of the content (r=-0.741**), soil pH (r=-0.535**) and investigated soils is shown in Table (10). then soil ECe (r=-0.343*).These relations are in Generally, the extractable Ni is slightly higher in agreement with those obtained by Roshdy (2009). the youngest Nile terrace soils than in the oldest Concerning the relation between the DTPA- ones with a rang of 0.38 to 1.52 ppm, without any extractable Ni and its bearing minerals i.e., clear trend of its distribution with depth (Table 10 pyroxenes, hornblende, garnet and biotite (sand and Figure 5). The lower values of the DTPA- fraction) as well as sepiolite and chlorite (Duane extractable Ni are recorded in the terrace bench and Robert, 1989), correlation coefficients were in the interference zone and the terrace rear calculated between them. Table (12) clearly shows suture in desert fringes on both sides, where they highly significant positive correlations between range from nil to 0.39 ppm. These soils do not the extractable Ni with both biotite (r=+0.648**) have also any specific pattern of distribution with and chlorite (r=+0.453**) as well as a slightly depth (Figure 5). These results agree with those of significant positive correlation with hornblende Abdel-Mawgoud and Faragallah (2004), (r=+0.351*), indicating that these minerals may Faragallah and Essa (2005) and Roshdy (2009). be as a source of extractable nickel in the studied Correlation coefficients calculated to test the soils. However, a highly significant negative relation between some soil properties and the correlation was observed between the extractable DTPA-extractable Ni, are listed in Table (11). The Ni and garnet (r= -0.467**). Pyroxenes showed a results show highly positive significant very low and negative correlation (r= -0.047) with correlations between the DTPA-extractable Ni the extractable Ni. and the soil contents of fine fractions (clay, silt and sum of silt+clay), organic matter, and CEC.

Table 10: DTPA-extractable Ni, Pb and Cd in the soil samples of the studied Nile terraces Profile Depth Ni Pb Cd Profile Depth Ni Pb Cd No. (cm) (ppm) No. (cm) (ppm) 0 - 15 0.62 0.34 0.01 0 - 20 1.36 0.71 0.02 15 - 25 0.78 0.36 0.02 5 20 - 35 1.04 0.74 0.02 1 25 - 35 0.47 0.34 0.02 35 - 65 0.96 0.24 0.00 35 - 70 0.67 0.44 0.02 0 - 45 1.52 0.75 0.02 6 0 - 40 0.72 0.43 0.02 45 - 80 1.07 0.24 0.00 40 - 70 0.86 0.21 0.01 0 - 40 0.39 0.20 0.00 2 70 - 95 0.75 0.30 0.01 40 - 70 0.52 0.27 0.02 7 95 - 195 0.82 0.33 0.02 70 - 120 0.38 0.02 0.00 195-235 0.95 0.37 0.03 120-155 0.55 0.37 0.02 0 - 35 0.39 0.63 0.01 0 - 50 0.27 0.00 0.00 35 - 55 0.06 0.64 0.00 50 - 70 0.05 0.21 0.01 3 8 55 - 70 0.11 0.61 0.04 70 - 85 0.00 0.04 0.00 70 - 150 0.07 0.38 0.05 85 - 150 0.00 0.09 0.00 0 - 20 0.00 0.00 0.00 0 - 10 0.21 0.58 0.00 4 20 - 35 0.07 0.37 0.02 9 10 -35 0.00 0.11 0.00 35 - 150 0.00 0.24 0.01 35 - 150 0.03 0.00 0.00

-85- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Profile 1 Profile 5

1 1.6 1.4 0.8 1.2 Ni Ni 0.6 1 Pb 0.8 Pb 0.4 Cd 0.6 Cd 0.4 0.2

0.2 Concentration (ppm) Concentration Concentration (ppm) Concentration 0 0 1 2 3 4 1 2 3 Layer No Layer No

Profile 6

1.6 1.4 1.2 1 Ni Profile 2 0.8 Pb 0.6 Cd 0.4 0.2

1 (ppm) Concentration 0 0.8 1 2 3 Ni Layer No 0.6 Pb 0.4 Cd Profile 7 0.2

Concentration (ppm) Concentration 0.6 0 0.5 1 2 3 4 5 0.4 Ni Layer No 0.3 Pb 0.2 Cd 0.1 Concentration (ppm) Concentration 0 1 2 3 4 Layer No

Profile 3 Profile 8

0.7 0.3 0.6 0.25 0.5 Ni 0.2 Ni 0.4 Pb 0.15 Pb 0.3 Cd 0.1 Cd 0.2

0.1 0.05 Concentration (ppm) Concentration Concentration (ppm) Concentration 0 0 1 2 3 4 1 2 3 4 Layer No Layer No

Profile 4 Profile 9

0.4 0.7 0.35 0.6 0.3 0.5 0.25 Ni Ni 0.4 0.2 Pb Pb 0.3 0.15 Cd Cd 0.1 0.2

0.05 0.1 Concentration (ppm) Concentration Concentration (ppm) Concentration 0 0 1 2 3 1 2 3 Layer No Layer No

Fig. 5: Distribution of extractable Ni, Pb and Cd with depth in the soil profiles of the studied Nile terraces

-86- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 11: Correlation coefficients of DTPA-extractable Ni, Pb and Cd and some physical and chemical properties of the studied soils r value Soil property Ni Pb Cd Sand (%) -0.741** -0.381* -0.143 Silt (%) +0.605** +0.321* +0.132 Clay (%) +0.729** +0.362* +0.120 Silt + Clay (%) +0.741** +0.381* +0.143 Organic matter (%) +0.675** +0.199 +0.052 * CaCO3 (%) -0.801** -0.332 -0.229 pH (1:1) -0.535** -0.165 +0.029 -1 ECe (dsm ) -0.343* -0.146 -0.212 CEC (Cmol+/kg) +0.720** +0.381* +0.289

Table 12: Correlation coefficients between Ni and Pb and their bearing-minerals Element Mineral r value Element Mineral r value Hornblende +0.351* Opaques -0.484** Pyroxenes -0.047 Amphiboles +0.375* Botite +0.648** Botite +0.117 Ni Pb Grnet -0.467** Fldspars (sand fraction) +0.131 Clorite +0.453** Fldspars (clay fraction) +0.172 Spiolite -0.193 * Significant at the 5% level. ** Significant at the 1% level.

2-Lead: The correlation coefficients in Table (11) clearly indicate positive significant correlations Table (10) reveals that the DTPA- between the DTPA-extractable Pb and the soil extractable Pb of the studied soils varies from contents of silt (r=+0.321*), clay (r=+0.362*) and 0.0 to 0.75 ppm. The youngest Nile terraces soils sum of silt+clay (r=+0.381*) as well as CEC contain a higher concentration of the extractable (r=+0.381*). On the other hand, negative Pb than the oldest ones, with a range of 0.24 to significant correlations were obtained between the 0.75 ppm and 0.02 to 0.43 ppm, respectively. DTPA-extractable Pb and both sand (r=-0.381*) Although most soil samples of the terrace bench and CaCO (r=-0.332*) contents. The data also in the interference zone and the terrace rear 3 show low correlations between the extractable Pb suture in desert fringes on both sides have levels and organic matter content (r=+0.199), soil pH of the extractable Pb ranging from 0.0 to 0.38 (r=-0.165) and soil EC (-0.146). Similar relations ppm, the first three layers of profile 3 and the e were found by Marto-Anez and Motto (2000) and surface layer of profile 9 display values between Roshdy (2009). 0.58 and 0.64 ppm. Like extractable Ni, Lead occurs in feldspars, biotite, extractable Pb concentrations show no trend of amphiboles, quartz and magnetite (Wedepohl, change with depth (Figure 5). Similar results were 1974). Accordingly, correlation coefficients were reported by Abdel-Mawgoud and Faragallah calculated between these minerals and the DTPA- (2004), Faragallah and Essa (2005) and Roshdy extractable Pb (Table 12) to verify the source of (2009).

-87- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

lead in the investigated soil. The obtained results obtained by Ghallab and Usman (2007) and reveal a significant positive correlation between Roshdy (2009). the extractable Pb and amphiboles (r=+0.375*) Cadmium occurs in salirite mineral which is as well as low positive correlations with biotite extremely rare in soils. Therefore very much (r=+0.117), feldspars of the sand fraction lower of cadmium amounts that is detected in (r=+0.131) and feldspars of the clay fraction the studied Nile terraces due to anthropogenic (r=+0.172), suggesting these minerals are contamination. This finding is in full agreement considered a source of the extractable Pb in the with those found by Zhou et al. (2004), Jjemba studied soils. A highly significant negative (2005) and Jinling et al. (2009). correlation between the extractable Pb and opaques (r=-0.484**) was observed (Table 12). REFERENCES:

Abdellah, K. (1984): The status of certain 3-Cadmium: micronutrients in soil representative soils Compared to the above mentioned heavy of Upper Egypt. M.S. Thesis, Faculty of metals, the DTPA-extractable Cd concentration Agric., Assiut Univ., Egypt. is very much lower in all studied soils and Abdel-Mawgoud, A.S.A. and M.A. Faragallah ranges from 0.0 to 0.05 ppm (Table 10). The (2004): Characterization of Some Nile youngest and oldest Nile terrace soils contain Alluvial Soils at Assiut, Egypt. J. Agric. similar amounts of the extractable Cd with a Sci. Mansoura Univ., 29 (9): 5335-5345. range of 0.0 to 0.03 ppm. Most soil samples of the Abd El-Razek, M., M.F. Ghoneim, G.S. El- terrace bench in the interference zone and Gharably and K. Abdellah (1984): the terrace rear suture in desert fringes on both Micronutrient status in soils of Upper sides do not have extractable Cd. Like Egypt: 4- Manganese. Assiut J. Agric. Sci. extractable Ni and Pb, the distribution of Cd in 15: 51-62. all location under study with depth is Abou El-Roos, S.A. (1972): Status of some unsystematic (Figure 5). macronutrient elements in Egyptian soils. Table (11) shows that, in all cases, the Ph. D. Thesis, College of Agric., Cairo correlation values between the concentration of Univ., Egypt. the DTPA-extractable Cd and the considered Amer, A.H., F.E. Abu-Agwa, and A.S. Awaga soil properties are low and insignificant. The (1991): Distribution of nutrients and their correlation coefficient values are positive with relations to some properties of selected soil contents of silt, clay, silt+clay, organic Egyptian soils. Minufiya J. Agric. Res. 16: matter, pH and CEC. However, they are 1015-1027. negative with soil contents of sand, CaCO3 and Attia, K.K. (1988): Studies on certain ECe. Most of these findings coincide with those micronutrients in the soils of Aswan and

-88- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

El-Minia governorates. M.Sc. Thesis, industrial area near Assiut city. Ass. College of Agric., Assiut Univ., Egypt. Univ. Environ. Res. 3 (1): 1-19. Awadalla, A.A.(1993): Mineralogical and physical El-Hadedy, E.M. (1976): Effect of irrigation on properties of some soils at El-Fayoum fertility level of Upper Egypt Soils after the governorate and their relation to soil construction of the High Dam. M.Sc. fertility. M.Sc. Thesis, Faculty of Agric., Thesis, Fac. of Agric., Assiut Univ., Egypt. Cairo Univ., Egypt. El-Shazly, M.M., E.A.A. Hamid, A.A. Moussa Brewer, R. (1964): Fabric and Mineral Analysis and M.A. Metwally (1991): Iron and of Soils. John Willey & Sons Inc, New manganese status in some Torrerts and York, London, Sydney. Fluvents, Egypt. Egyptian J. Soil Sci. 31: Chakrapani, G. J. (2005): Factors controlling 115-130. variations in river sediment loads. Faghami, A., S.O. Ajayi and E.A. Ali (1985): Current Sci., 88 (4): 569-575. Nutrient distribution in the basement Cooke, G.W. (1967): The control of soil fertility. complex south western Nigeria: 1- Crosby Lockwood, London. Macronutrients - calcium, magnesium and Dekov, V.M., Z. Komyb, F. Arahjo’, A. Van potassium. Soil Sci. 139: 431-438. Putd and R. Van Griekend (1997): Farragallah, M. A. (1995): Relative distribution Chemical composition of sediments, of certain nutrients in soils of the Nile suspended matter, river water and valley-desert interference zone east of ground water of the Nile (Aswan-Sohag Assiut city. M.Sc. Thesis, College of traverse). The Science of the Total Agric. Assiut Univ., Egypt. Environment 201: 195-210. Faragallah, M.A. and M.A. Essa. (2005): Dress, L.R. and L.P. Wilding (1978): Elemental Physical, chemical and macro- distribution in the light mineral isolate of micromorphological characteristics of soil seperates. Soil Sci. Am. J. 42: 976-978. some alluvial soils irrigated with different Duane, M. and C. R. Robert (1989): X-ray water resources. Assiut, Univ. Bull. diffraction and the identification and Environ. Res. 8(1): 51-65. analysis of clay minerals. Oxford Univ. Ghallab, A. (2002): Pollution status of soils and Press, Inc. Madison Avenue, New York, plants around the factory of ferrosilicon USA. alloys, Edfo, Aswan. Ass. Univ. Environ. Eden, D.N. and R.L. Parfitt (1992): Amounts and Res. 5 (1): 33-54. sources of phosphate in hill country rocks, Ghallab, A. and A. R. Usman (2007): Effect of south-eastern North Island, New Zealand. sodium chloride-induced salinity on Australian J. Soil Res. 30: 357-370. phyto-availability and speciation of Cd in El-Desoky, M.A. and A. Ghallab (2000): soil solution. Water air soil Pollut, 185: Pollution impact on soil and plants in an 43-51.

-89- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ghoneim, M.F., M. Abd El-Razek, G.S. El- Jjemba, P. K. (2005): Bioavailability of metals Gharably and K. Abdellah (1984): and metalloids in terrestrial Micronutrients status in soil of upper environments p 25-41. In Ahmed, I., S. Egypt:1- Zinc. Assiut J. Agric. Sci. 15:15- Hayat and J. Pichtel (eds) Heavy metal 26. contamination of soil. Problems and Gladilovich, B.R. and Zh.V. Sergeeva (1971): remedies. Sci. Pub., Inc., Enfield, NH, Effect of mineralogical composition on USA. the level of trace elements in some soil- Kabata-Pendias, A. and H. Pendias (1984): Trace forming rock of the Leningrad district. elements in soils and plants. CRC Press, Zapiski Leningrado Shogosel Inc., Boca, Florida, USA. Skokhozaistvennogo Instuta 160: 10-15. Khalifa, E.M., M. A. El-Desoky, M. A. Gameh (C.F. Soils and Fert. 35:645). and M. A. Faragallah (1996): Status of Gomah, H. H., M. AbdElRazek, M. A. El- some micronutrients and their relations to Desoky and A. A. Amin (2007): Status of mineral composition of the Nile valley- iron and forms in soils of Assiut. J. Agric. desert interference zone east of Assiut Sci. Mansoura Univ., 32 (5): 4107-4119. city. Assiut J. Agric. Sci. 27(3):107-127. Jackson, M. L. (1973): Soil Chemical Analysis. Khalil, I.K., M. M. Gundy, A.S. Metwally and Prentice Hall Inc., New Delhi. M.M. Fahim (1990): Manganese status in Jakovljevic, M. D., N. M. Kostic, D. Stevanovic , some soils of Sinai. Zagazig J. Agric. Res. S. Blagojevic, M. J. Wilson and Lj. 17: 31-40. Martinovic. (1997): Factors influencing Khetawat, G.K. and K.S. Vashishtha (1977): the distribution of heavy metals in the Distribution of micronutrients and their alluvial soils of the Velika Morava River relationship with some soil properties in valley, Serbia. Applied Geochemistry. 12: the vineyards of Rajathan. J. Indian Soc. 637-642. Soil Sci. 25: 81-83. Jinling, L., H. Ming, H. Wei and G. Yifan Li, X. and I. Thornton (2001): Chemical (2009): Analysis and assessment on heavy partitioning of trace and major elements in metal sources in the coastal soils soils contaminated by mining and smelting developed from alluvial deposits using activities. Appl. Geochem. 16: 1693–1706. multivariate statistical methods. J. Lindsay, W.L. and W.A. Norvell. (1978): Hazardous Materials 164: 976–981 Development of DTPA soil test for zinc, Jjemba, P.K. (2004): Environmental iron, manganese and copper. Soil Sci. Soc. microbiology: Principles and Am. J. 42: 421-428. Applications. Science Publishers, Inc., Marto-Anez, C. E. and H. L. Motto (2000):. Enfield New Hampshire, USA. Solubility of lead, zinc and copper added

-90- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

to mineral soils. Environmental Pollution contamination of air, water and soils by 107: 153-158. trace metals, Nature 333: 134–139. McLean, E.O. (1982): Soil pH and lime Panuccio, M.R., A. Sorgonà, M. Rizzo and G. requirement. P. 199-224 In A.L. Page, Cacco (2009): Cadmium adsorption on R.H. Miller and D.R. Keeney (eds). vermiculite, zeolite and pumice: batch Methods of soil analysis, part 2. Chemical experimental studies. J. Environ. Manage and microbiological properties, 2nd 90 (1): 364–374. edition. Soil Sci. Soc. Am. Inc., Madison, Peijnenburg, W.J.G.M. and T. Jager, (2003): WI, USA. Monitoring approaches to assess Metson, A. J. (1961): Methods of chemical bioaccessibility and bioavailability of analysis for soil survey samples. New metals: matrix issues. Ecotoxicol. Zealand Dept. Sci., Ind. Res. Soil. Bur. Environ. Saf. 56 (1), 63–77. bull., 12 Govt. Printer, Wellington, Zew Piper, C.S. (1950): Soil and plants analysis. Zealand. (C.F. Booker Tropical Soil Inter. Science, Inc., New York. Manual by London, J.R., 1984, p. 126- Rabie, F., A. El-Araby and M. El-Shazly. 1989. 138). Micronutrients status as related to soil Milner,H.B.(1962): Sedimentology petrography. constituents in soils of Eastern Desert, II & I. George Allen & Unwin, LTD. Egypt. Egyptian J. Soil Sci. 29: 331-343. London. Roshdy, N. M. K. (2009): Distribution and Mitchell, R.L. (1964): Trace elements in soils p. forms of some heavy metals in a 320-368. In F.E. Bear (ed.) Chemistry of contaminated soil at Assiut. Ph.D. Thesis, the soils. Reinhold publishing cooperation, Fac. of Agric., Assiut Univ., Egypt. New York, USA. Sillanpaa, M. (1972): Trace elements in soils Mohamed, N.A. (1990): Nutrient status in some and agriculture. FAO Soils Bull. 17, soils as related to mineralogical Rome, Italy. composition. M.Sc. Thesis, Faculty of Schultz, L.G.(1964): Quantitative interpretation Agric., Ain Shams Univ., Egypt. of mineralogical composition from X-ray Nelson, R. E. (1982): Carbonate and gypsum. P. and chemical data for the Pierre shale. 181-198 In A. L. Page, R.H. Miller and D. U.S. Geol. Surv., Prof. Paper, No.391-C, R. Keeney (eds). Methods of soil analysis. pp.C1-C31, Washington. Part 2, Chemical and microbiological Soltanpour, P.N. and A.R. Schwab (1977): A properties, 2nd edition. ASA and Soil Sci. new soil test for simultaneous extraction Soc. Am. Inc., Madison, WI, USA. of macro- and micro-nutrients in alkaline Nriagu, J.O. and J.M. Pacyna (1988): soils. Comm. in Soil Sci. and Plant Anal. Quantitative assessment of worldwide 8, 195.

-91- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Stehouwer, R. C., A. M. Wolf and W. T. Doty composting on characteristics of humic (2000): Chemical monitoring of sewage acid. Chemosphere 68, 368–374. sludge in Pennsylvania: Variability and White, M. L. (1957): The occurrence of zinc in application uncertainty. J. Environ. soil. Econ. Geol. 52: 645-651. Quality 29: 1686-1695. Wiklander, L. (1958): General occurrence of Tessier, A. and P. G. C. Campbell (1988): plant nutrients. In Encyclopedia of plant Partitioning of trace metals in sediments. physiology: V. Mineral nutrition of plants. P. 183–199. In Kramer J. R and H. E. Springer Verlage, Berlin. Allen (eds). Theory, analysis and Yadav, S.K., A. A. Juwarkar, G.P. Kumar, P.R., application. Chelsea7 Lewis Publishers. Thawale, S.K. Singh and T. Chakrabarti Viets, F.G. and W.L. Lindsay (1973): Testing soil (2009): Bioaccumulation and phyto- for Zn, Cu, Mn and Fe. p. 153-172. In. translocation of arsenic, chromium and L.M. Walsh, and J.D. Beaton. Soil Testing and Plant analysis. Soil Sci. Soc. Am. Inc., zinc by Jatropha curcas L.: Impact of Madison, WI, USA. dairy sludge and biofertilizer. Bioresour. Velde, B. (1992): Introduction to clay minerals. Technol. 100 (20): 4616–4622. Chapman & Hall, London, Glasgow, New Zhou, H. Y., X. T. Penga and J.M. Panb. (2004): York, Tokyo, Melbourne, Madras. Distribution, source and enrichment of Wedepohl, K. H. (1974): Handbook of some chemical elements in sediments of Geochemistry. Springer-Verlag, Berlin, the Pearl River Estuary, China. Heidelberg, New York, 839 P. Continental Shelf Research 24: 1857– Wei, Z., B. Xi, Y. Zhao, S. Wang, H. Liu and Y. 1875.

Jiang (2007): Effect of inoculating microbes in municipal solid waste

-92- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

حالة بعض العناصر الغذائية والعناصر الثقيلة وعالقاتها بالمعادن الحاملة لها فى أراضى المدرجات النهرية بأسيوط - مصر محمد األمين عبد الكريم فرج هللا

قسم علوم األراضى والمياه – كلية الزراعة – جامعة األزهر –أسيوط .

تم اختيار تسع قطاعات تمثل أراضى المدرجات النهرية الحديثة والقديمة المتكونة فى منطقة جنوو قنواطر أسويوط وذلو لدراسوووة مسوووتويات وتوزيوووع عوووا العناذووور ال ذا يوووة )Cu ،Zn ،Mn ،Fe ،K ،P( و عوووا العناذووور الثقيلوووة )Cd ،Pb ،Ni( وعالقاتها عا الخواص الط يعية والكيميا ية لهذه األراضى، وكذل لتقييم ارت اط هذه العناذور موع المعواد الحاملوة لهوا فوى هوذه األراضى. وقوود أرهوورت النتووا ح المتحذوول عليهووا أ قوويم الفوسووفور الميسوور كووا أعلووى فووى أراضووى الموودرجات النهريووة القديمووة منوو فووى المدرجات الحديثة على جان ى النيل، ينما محتوى ال وتاسيوم الميسر أرهر العكس. وقد احتوت أراضوى المودرجات النهريوة الحديثوة والقديمة على مستويات عالية م الفوسفور و مستويات متوسطة م ال وتاسيوم . ولم يوجد ت ير ثا ت فى توزيع كل م الفوسفور وال وتاسيوم مع العمق فى معرم القطاعات. وقد ُوجد ارت اطات موج ة معنوية جداً ي تركيوزات كول مو الفوسوفور وال وتاسويوم موع محتوى هذه األراضى م السلت والطي والمادة العضوية والسعة الت ادلية الكاتيونية، ولك كا االرت اط سال ا ً مع محتوى التر ة م الرمل وكر ونات الكالسويوم والوو pH. كموا اتضو وجوود ارت اطوات موج وة معنويوة وي محتووى الفوسوفور الميسور موع كميوات معود المونازيت ومجموع معدنى األ اتيت والمونازيت. كما يوجد ارت اط موج عالي ي ال وتاسيوم ومعاد الفلس ار التى فى حجم الطي لهذه األراضى. أوضحت النتا ح أيضوا أ المسوتويات األعلوى مو الحديود الميسور كانوت فوى أراضوى المودرجات النهريوة القديموة، ينموا محتووى المنجنيز الميسر كا أعلى فى أراضى المدرجات الحديثوة، ولوم يُرهور عنذور النحواس ت يو ارت ك يورة وي أ ارضوى المودرجات القديموة والحديثة. واحتوت أراضى كل المودرجات النهريوة علوى مسوتويات كافيوة مو الحديود والمنجنيوز والنحواس، ينموا أرهورت كول األراضوى نقذا ً فى الزن الميسر. كما رهرت ارت اطات موج ة عاليوة وي الحديود والمنجنيوز والنحواس موع النسو ة الم ويوة للح ي وات الناعموة خاذة الطي ونس ة الموادة العضووية والسوعة الت ادليوة الكاتيونيوة ل راضوى، وأخورى سوال ة موع محتووى التر وة مو الرمول وكر ونوات الكالسيوم والو pH، ينما ال توجد ارت اطات معنوية ي عنذر الزن وأى م خواص التر ة المدروسة. كما توجد ارت اطوات موج وة عالية ي كل م الحديد والنحاس م جهة والمعاد الحاملة لها قليلة الث ات مثل اال يدوت وال يوتيوت والفلسو ار مو جهوة أخورى، و ي المنجنيز ومعاد اال يدوت والهورن لند والسمكتيت والكلوريت. ينما توجد ارت اطات موج ة منخفضة ي عنذر الزن وكل م الهورن لند وال يروكس .

ّينت النتا ح أ مستوى عنذرى النيكل والرذاص كا أعلوى قلويالً فوى أ ارضوى المودرجات النهريوة القديموة منو فوى المودرجات الحديثة على جان ى النيل و دو ت ير واض مع العمق ، ينما كا مسوتوى عنذور الكوادميوم متهوا ومونخفا جوداً فوى كول مو أراضى المدرجات النهرية الحديثة والقديمة. وقد وجد ارت اط موج معنوى عوالى وي كميوة النيكول المسوتخلص موع النسو ة الم ويوة للسوولت والطووي ونسوو ة المووادة العضوووية والسووعة الت ادليووة الكاتيونيووة ل راضووى. ورهوور ارت وواط موجوو معنوووى ووي كميووة الرذوواص المسوتخلص موع النسو ة الم ويوة للسولت والطوي والسوعة الت ادليوة الكاتيونيوة، ورخور سوال موع محتووى التر وة مو الرمول وكر ونووات الكالسيوم. وقد أتض وجود ارت اطات موج ة معنوية درجة منخفضة ي كل م عنذرى الرذاص والنيكل ومعد الهورن لند ينما ارت اط موج عالى ي عنذر النيكل وكل م معدنى ال يوتيت والكلوريت .

-93- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

MINERALOGICAL COMPOSITION OF KHAMASIN WIND DUST AT ASSIUT, EGYPT Mohamed El-Ameen A. Farragallah* and Mahmoud A. Essa** *Soil and water Sic. Dept., Faculty of Agric., Al-Azhar University, Assiut, Egypt **Geology Dept., Fac. Sci. Assiut University, Assiut, Egypt.

ABSTRACT: The present study investigated the mineralogical composition of the deposited dust from Khamasin wind on the agricultural soils at Assiut city through April 2008 and 2009. Five sites were chosen between the fringes of eastern and western deserts throughout the Nile valley. Wood boxes (dimensions of 1x1m) were put in the selected sites and precipitated dust was collected from each box after the finish of Khamasin storms in the two years. The quantity, pH and EC were determined for the collected Khamasin dust. The mineral particles of these dusts were examined by the scanning electron microscope (SEM) and the X-ray analysis. The obtained results revealed some variation in the dust quantity of Khamasin wind in the year of 2008 and 2009. The quantity of the collected bulk samples of the Khamasin dust varied from site to another in both 2008 and 2009 years. The highest amounts in both years are recorded in the sites of the eastern side of the Nile valley that faraway from the western desert, where the Khamasin wind come. The mean values of dust EC and pH were low and similar in both years of 2008 and 2009. The Scanning electron microscope photomicrographs showed that the grain sizes of Khamasin dust in the sites located in the interference zone of Nile valley with the eastern desert were finer than those in site located in the western side of the Nile river. Generally, relatively large angular to subangular grains, larger amount of very fine grains with more oblate particle and spherical particles with differences in size as well as particles with irregular shapes in Khamasin dust of 2008 and 2009 years. Semi-quantitative measurements of the identified minerals in the Khamasin dust of 2008 year have shown the order of quartz>k-feldspar>calcite>magnetite>epidote>anhydrite>garnet> augite> tourmaline >zircon>ilmenite>plagioclase>goethite>actinolite>hematite. However, the minerals in the Khamasin dust of 2009 year could be ordered as quartz>k-

feldspar>calcite>epidote>magnetite> tourmaline >anhydrite >augite>plagioclase >ilmenite>rutile>hematite>garnet>actinolite. There was no general trend for distribution of these minerals throughout the studied sites. From a qualitative point of view, most of the sites have the same mineralogical composition. Comparison of the mineralogical compositions of mineral dusts at the five sites, revealed certain quantitative differences.

-95- INTRODUCTION: within the source regions with high illite and low kaolinite+chlorite contents. The The recognition and characterization of mineralogical composition of dust collected in aeolian dust in soil contribute to a better the deposition area of the Chinese Loess Plateau understanding of landscape and ecosystem (Changwu) was enriched by clay minerals (Shen dynamics of drylands (Reynolds et al., 2006). et al., 2009). Soil dust is recognized as an important factor on Khamasin winds of Egypt and North Africa, climate forcing (Duce, 1995; Li et al., 1996; which coming from the south-east and south, Arimoto, 2001; Arimoto et al., 2006). Aeolian carry the Libyan dust over the Mediterranean. dust influences surficial and ecological processes With deep depressions of the western desert of in desert regions, where large amounts of dust Egypt, Khamasin winds blow with great are both sequestered and emitted over long violence. In sweeping the desert to the south, periods of time (Goudie, 1978). In these settings, they have a double action. Being hot and dry, eolian dust plays important roles in soil with relative humidities of 30, 20, even 10 formation, soil hydrology, development of percent., the mantle of dust is fully dried out so surfaces, distribution of biologic soil crusts and that the grains lose all coherence, and in this nutrient status (Chadwick and Davis, 1990; desiccated state are raised freely into the air and McDonald et al., 1996; Belnap and Gillette, carried along as ust-clouds. In its southerly 1998; McFadden et al., 1998; Shachak and phase the Khamasin brings temperatures Lovett, 1998; Reynolds et al., 2001). Mineralogy of aerosols has been used to approaching 40C. (104F.), but when it veers to assess the sources of eolian sediments (Leinen et the north-west there is rapid cooling and within al., 1994; Merrill et al., 1994; Biscaye et al., fifteen minutes the temperature reading may 1997; Drab et al., 2002; Bory et al., 2002). drop to 27C. (80.6F.). These hot winds Mineralogy may be more diagnostically characterize the later months (April-May): significant for identifying aerosol source areas earlier in the year a Khamasin may be relatively than elemental ratios (Schutz & Sebert, 1987 cold. But even after the wind has veered north- and Molinaroli et al., 1993). Clay mineral ratios, west the dust-storm will continue till the velocity such as I/K ratio (I: illite, K: kaolinite) can also abates. The visibility at times being restricted to be used as relevant tracers of the origin of soil- only a few metres, whilst in exposed desert derived aerosols within Saharan source regions places it was reported as nil and the velocity of (Caquineau et al., 1998). The main the wind reached 53 miles per hour in some mineralogical difference in dust composition is places (Oliver, 1945). its carbonate (especially calcite) and feldspar The highest average values of dust deposit, contents. On the other hand, the clay all over the greater Cairo city, were found in the mineralogy of the dust displays little variation spring, which is characterized by the passage of

-96- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

the hot Khamasin storms that are loaded with March through May, that brings very large dust and sand (Hgazy, 1961). Deposited dust amounts of sand and even dust from the resulting from such storms varied in the mean Arabian Desert to cities and the agricultural from 15.6 to 23.1 ton/mile2/storm (Salam, and soils. Khamasin blows from the south to the Sowelim, 1967). However, the average rate of northwest, in opposition to the prevailing winds. dust deposit in the industrial center at Helwan, Egypt is mostly desert, with the exception of the about 20 km south of Cairo, for the month of Nile River delta and basin. As a result, the April, when Khamasin dust storms were more country is hot and dry most of the year-round. frequent, was found to be 117.6 ton/mile2 in Egypt experiences very mild winters. In the 1967 (Salam, 1967). The silt-plus-clay fraction spring, the country is subject to a hot, driving (fines) of the recently deposited aeolian dust on windstorm called the Khamasin. The Khamasin the isolated surfaces is closely alike in magnetic- causes major drifting of sand. Due to its desert mineral composition, in contrast to greatly terrain, Egypt is also prone to dust and dissimilar magnetic compositions of rock sandstorms, along with periodic droughts. Sand surfaces of vastly different lithologies, on which storms are very dangerous for residents because the fines have accumulated. The fines, thus, are they can continue for days, making life predominantly deposited dust. The extremely difficult for citizens to live. Illness can compositional similarity of the dust on these occur from sand storms as a result of days of surfaces is interpreted to result from mixing of brutal winds and even damage to homes, fines in the atmosphere as well as in fluvial, infrastructures and crops can leave local alluvial, and lacustrine depositional settings Egyptians with a sense of unease and sadness. prior to dust emission (Reynolds et al., 2006). In Assiut, Khamasin usually arrives in April No research on the particulate air dust of every year carrying great quantities of dust and Khamasin wind around Assiut has been sand from the deserts, which shrouded the city documented. Therefore, the present study was in a thick yellowish haze. Khamasin wind dust carried out to investigate the quantity, some in Assiut increase in particulate air matter chemical properties and mineralogical considered as environmental contaminants. composition of the deposited dust from Metrological data reveal that the prevailing Khamasin wind on the agricultural soils at climate is corresponding to hot desert; the Assiut city. maximum temperature varies from 21C in January to 41C in July and the minimum MATERIALS AND METHODS: temperature varies from 4.5C in January to Khamasin is a Middle Eastern term for the 22C in July. The average value of high relative dry, hot wind that blows in from the desert. It humidity is about 78.92% while the low one is can refer to the wind that blows from the desert about 18.17%. Pan evaporation is about 1910.2 across Egypt in the spring, typically from

-97- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

mm per year. Precipitation is practically nil, 2009. The precipitated dust was collected from except some light showers of rain that rarely fall each box after the finish of Khamasin storms in during winter and some unrecorded flush floods the two years. The quantity of each sample was that come from the Eastern Desert (Faragallah, weighted to calculate the average of annual 1995; Ainer and Eid, 2003). addition of Khamasin dust to the agricultural To study the annual additions and the soils at Assiut City. The electrical conductivity mineralogical composition of the Khamasin (EC) of the collected dust was measured in 1:2.5 dust, five sites were chosen between the fringes soil to water extract by using a conductivity of eastern and western deserts throughout the meter. The pH was measured in 1:2.5 soil: water Nile valley as shown in Figure (1). Wood boxes suspension using a glass electrode as reported with dimensions of 1x1m were put in the soil by Mclean (1982). surface of these sites in the 1st of April 2008 and

Figure 1: Locations of the wood boxes for collection the deposited Khamasin dust

The bulk samples of the collected Khamasin grain particles were examined by using dust were treated to remove the soluble salts Scanning Electron Microscope (SEM). The and organic matter for investigation by X-ray samples were prepared by mounting onto Diffractometer (XRD) and the scanning electron aluminum stubs and then coating with gold for microscope (SEM). The sizes and shapes of the best resolution using JEOL-JSM-5400 LV SEM.

-98- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

The dust samples were ground for examination quantity of deposited Khamasin dust in 2009 by X-ray Diffractometer (XRD). The mineral varied between 1.0129 and 5.4678 g/m2. The phases identified by X-ray powder diffraction highest amounts in both years are recorded in technique. The powder samples were the sites of the eastern side of the Nile valley investigated using PHILLIPS X-ray that faraway from the western desert, from diffractometer with CuKα radiation, 45 KV and where the Khamasin wind come. This may be 35 mA and scanning between 2θ of 2 and 60. due to the decrease of Khamasin wind velocity The mineral constituents of the studied samples then loses most their carriage. The mean were identified and their relative proportions quantity of the bulk samples of the deposited were determined using the semi-quantitative Khamasin dust varied from year to another, peak area technique according to Schultz since the mean quantity is higher in the 2009 (1964), Jackson (1964), Whitting (1965), Mac (2.4304 g/m2) than 2008 (2.3004 g/m2). This may Ewan (1968) and Carroll (1970). be attributed to the difference of velocity and the carriage of dust from year to another. RESULTS AND DISCUSSION: The electrical conductivity (EC) of the 1-The quantity and some chemical collected Khamasin dust in 2008 and 2009 years properties: was extremely low and ranged from 13.51 to -1 -1 The obtained data in Table (1) reveal that 14.08 µSm and 13.46 to 13.87 µSm , the quantity of the collected bulk samples of the respectively. The similarity of EC values in both Khamasin dust varied from site to another in 2008 and 2009 years may be due to the same both 2008 and 2009 years. Deposited dust source of dust. Also, the mean values of dust pH resulting from Khamasin storm in 2008 ranged were similar in year of 2008 and 2009 (6.20 and from 0.5106 to 4.8798 g/m2. However, the 6.19, respectively).

Table 1: The quantity, EC and pH of the Khamasin dust in the studied sites the quantity (g/m2) EC (µSm-1) (1:2.5) pH (1:2.5) Site No. 2008 2009 2008 2009 2008 2009 1 0.9208 1.0129 13.60 13.71 6.14 6.13 2 0.5106 1.0186 13.51 13.46 6.13 6.13 3 4.8798 5.4678 13.62 13.72 6.13 6.13 4 4.3954 4.9349 14.08 13.85 6.13 6.33 5 2.3004 2.4304 13.74 13.87 6.45 6.24 Mean 2.6014 2.9729 13.71 13.72 6.20 6.19

2-Dust mineralogy: A-The sizes and shapes of the mineral grains:

-99- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

The sizes and shapes of the grain particles site 2 in the western side of the Nile river. of Khamasin dust samples were examined by Generally, relatively large angular to using Scanning Electron Microscope (SEM) as subangular grains in Khamasin dust of 2008 shown in Figures 2 and 3. SEM observations and 2009 years, particularly, the finest fraction, provide more information about the finest dust often with elongated shape. A larger amount of particle. The Scanning electron microscope very fine grains with more oblate particle photomicrographs show that the grain sizes of shapes are observed. Spherical particles with Khamasin dust in site 4 were finer than those in differences in size as well as particles with site 2, where site 4 locate in the interference irregular shapes are present. zone of Nile valley with the eastern desert while

Figure 2: Scanning electron microscope photomicrograph showing the grain sizes and shapes of the Khamasin dust in site 2, (a) 2008 year and (b) 2009 year

Figure 3: Scanning electron microscope photomicrograph showing the grain sizes and shapes of the Khamasin dust in site 4, (a) 2008 year and (b) 2009 year

-100- B-The type and amount of minerals: amounts varied between 4.44 to 19.23% and X-ray powder diffractograms of the 4.76 to 25.18%, respectively (Tables 2&3). collected Khamasin dust from the studied sites Magnetite, epidote, anhydrite and garnet are shown in figures 4 and 5. Semi-quantitative were common minerals in Khamasin dust of measurements of the identified minerals in the 2008 year with the range reached up to 11.22, Khamasin dust of 2008 and 2009 years are given 6.31, 4.78 and 4.59%, respectively. However, in Tables 2 and 3, respectively. They have epidote, magnetite, tourmaline and anhydrite shown that the order of quartz>k-feldspar> were common minerals in Khamasin dust of calcite> magnetite>epidote> anhydrite> garnet> 2009 year with the range reached up to 9.35, 8.24, 4.43 and 5.06%, respectively (Tables 2&3). augite>tourmaline> zircon>ilmenite> plagioclase > goethite>actinolite> hematite in the Khamasin All of these minerals have not clear tend of dust of 2008 year. However, the minerals in the distribution with sites in both years (Figures Khamasin dust of 2009 year could be ordered as 6&7). quartz >k-feldspar>calcite>epidote> magnetite> The rest identified minerals i.e., augite, tourmaline> anhydrite> augite> plagioclase> tourmaline, zircon, ilmenite, plagioclase, ilmenite>rutile> hematite> garnet> actinolite. goethite, actinolite and hematite in the Quartz was the main constituent in the Khamasin dust of 2008 year only occurred in minerals of Khamasin dust samples with rang of few sites. Also, augite, plagioclase, ilmenite, 30.45-74.83% and 32.37-52.20% in years of rutile, hematite, garnet and actinolite were only 2008 and 2009, respectively (Tables 2&3). The found in few Khamasin dust samples of 2009 highest amounts of quartz (74.83 and 52.20%) year. for years of 2008 and 2009, respectively, are From a qualitative point of view, most the found in the site 5 that located in the fringe of sites have the same mineralogical composition. the eastern desert. Irregular pattern for the Comparison of the mineralogical compositions distribution of quartz content was observed in of mineral dusts at five sites, reveal certain both years (Figures 6&7). quantitative differences as shown in Tables 2&3 K-feldspar occurred as the second abundant and Figures 6&7. The carbonate content in Khamasin dust mineral in both 2008 and 2009 Khamasin dust increased from west to east in years, with amounts ranged from 11.56 to both two years. Similar to calcite, feldspar 23.36% and 9.49 to 23.10%, respectively. content increased from west to east up to site 4 Calcite present as the third abundant Khamasin then decreased. dust mineral in both 2008 and 2009 years, with

-101- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 2: Semi-quantitative measurements of the identified minerals in the Khamasin dust of 2008 year

ite

Site c

No.

feldspar

Augite

Zircon

agioclase

Cal

Quartz Garnet

-

Epidote

Ilmenite

Goethite

Hematite

Actinolite

Magnetite

Anhydrite

K

Pl Tourmaline

1 31.74 17.59 5.35 5.74 6.31 4.78 4.59 10.71 0.00 5.16 4.40 0.00 3.63 0.00 0.00

2 49.38 19.75 4.44 4.20 4.20 4.20 0.00 6.17 3.46 0.00 0.00 4.20 0.00 0.00 0.00

3 32.87 23.36 17.99 10.38 4.67 3.98 4.33 0.00 0.00 0.00 0.00 0.00 0.00 0.00 2.42

4 30.45 22.12 19.23 11.22 4.01 3.37 3.21 0.00 3.85 0.00 0.00 0.00 0.00 2.56 0.00

5 74.83 11.56 13.61 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00

Table 3: Semi-quantitative measurements of the identified minerals in the Khamasin dust of 2009 year

Site

No.

feldspar

Rutile

Augite

Calcite

Quartz Garnet

-

Epidote

Ilmenite

Hematite

Actinolite

Magnetite

Anhydrite

K

Plagioclase Tourmaline

1 36.50 9.49 5.49 5.27 4.85 4.43 5.06 9.70 4.22 3.59 8.02 3.38 0.00 0.00

2 47.06 17.37 4.76 4.20 4.76 3.92 4.20 6.44 4.20 0.00 0.00 0.00 3.08 0.00

3 32.37 19.78 25.18 9.35 3.96 3.06 3.96 0.00 0.00 2.34 0.00 0.00 0.00 0.00

4 34.48 23.10 24.83 5.86 4.83 3.62 3.28 0.00 0.00 0.00 0.00 0.00 0.00 0.00

5 52.20 13.46 17.58 3.02 8.24 3.02 0.00 0.00 0.00 0.00 0.00 0.00 0.00 2.47

-102- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Figure 4: X-ray diffractograms of the Khamasin dust of different sites in 2008 year

Figure 5: X-ray diffractograms of the Khamasin dust of different sites in 2009 year

-103- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

2008

80.00 Quartz 70.00 K-feldspar 60.00 50.00 Calcite Epidotes % 40.00 30.00 Magnetite 20.00 Anhydrite 10.00 Garnet 0.00 1 2 3 4 5 Site No.

Figure 6: Distribution of common minerals in the sites of Khamasin dust samples in 2008 year

2009

60.00 Quartz 50.00 K-feldspar 40.00 Calcite

% 30.00 Epidotes 20.00 Magnetite Tourmaline 10.00 Anhydrite 0.00 1 2 3 4 5 Site No.

Figure 7: Distribution of common minerals in the sites of Khamasin dust samples in 2009 year

-104- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

REFERENCES: tracer to assess the origin of soil derived aerosols. Geophysical Research Letters, Ainer, N.G. and H.M. Eid. (2003): Monthly 25(7): 983–986. Agro-metrological bulletin, Agric., Res. Carroll, D. (1970): Clay minerals. A guide to Center, General Administration for their X-ray identification. Geol. Soc. Am. Agricultural Instruction, No. 73 to 84. Spec. Paper 126. Arimoto, R. (2001): Eolian dust and climate: Chadwick, O.A. and J.O. Davis (1990): Soil- Relationships to sources, tropospheric forming intervals caused by eolian chemistry, transport and deposition. sediment pulses in the Lahontan basin, Earth-Science Reviews, 54: 29–42. northwestern Nevada. Geology 18, 243– Arimoto, R., Y. J. Kim, Y. P. Kim, P. K. Quinn, 246. T. S. Bates and T. L. Anderson (2006): Drab, E., A. Gaudichet, J. L. Jaffrezo and J. L. Characterization of Asian dust during Colin (2002): Mineral particles content in ACE-Asia. Global and Planetary change, recent snow at Summit (Greenland). 52:23–56. Atmospheric Environment,36:5365–5376. Belnap, J. and D.A. Gillette (1998): Duce, R. A. (1995): Sources, distributions and Vulnerability of desert biological soil fluxes of mineral aerosols and their crusts to wind erosion: the influences of relationship to climate. In R. J. Charlson, crust development, soil texture, and & J. Heintzenberg (Eds.), Aerosol forcing disturbance. Journal of Arid Environ- of climate (pp. 43–72). New York: John ments 39: 133–142. Wiley. Biscaye, P.E., F.E. Grousset, M. Revel, S. Van Faragallah, M. A. (1995): Relative distribution der Gaasr, G.A. Zielinski and A. Vaars of certain nutrients in soils of the Nile (1997): Asian provenance of glacial dust Valley-Desert interference zone, east of (stage 2) in GISP2 ice core, Summit, Assiut city. M.Sc. Thesis, Fac. of Agric., Greenland. Journal of Geophysical Assiut Univ., Egypt. Research, 102: 26765–26781. Goudie, A. S. (1978): Dust storms and their Bory, A.J.-M., P.E. Biscyae, A. Svensson and geomorphological implications. Journal F.E. Grousset (2002): Seasonal variability of Arid Environments 1: 291–310. in the origin of recent atmospheric Hgazy, N. A. (1961): Physical studies on mineral dust at northGRIP, Greenland. polluted air in Egypt. M. Sc. Thesis, Earth and Planetary Science Letters, 196: Cairo Univ. 123–134. Jackson, M.L. (1964): X-ray diffractogram Caquineau, S., A. Gaudichet, L. Gomes, M. C. interpretation. Chap. 8. In. C.L. Rich and Magonthier and B. Chatenet (1998): G.W. Kunze (eds.) Soil Clay Mineralogy. Saharan dust: Clay ratio as a relevant

-105- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Univ. North Carolina Press, Chapel Hill, edition. Soil Sci. Soc. Am. Inc., Madison, USA. WI, USA. Leinen, M., J.M. Prospero, E. Arnold and Merrill, J., E. Arnold, L. Margaret and C. Blank, M. (1994): Mineralogy of aeolian Weaver (1994): Mineralogy of aeolian dust reaching the North Pacific Ocean 1, dust reaching the North Pacific Ocean 2, sampling and analysis. Journal of Relationship of mineral assemblages to Geophysical Research, 99(D10): 21017– atmospheric transport patterns. Journal 21023. of Geophysical Research, 99(D10): Li, X., H. Marlng, D. Savole, K. Voss, and J. M. 21025–21032. Prospero (1996): Dominance of mineral Molinaroli, E., S. Guerzoni and G. Rampazzo. dust in aerosol light-scattering in the (1993): Contribution of Saharan dust to North Atlantic trade winds. Nature, 380: the central Mediterranean basin. 416–419. Geological Society of America, Special Mac Ewan, D.M.C. (1968): Types of paper, 284: 303–312. interstratification in soil clay minerals. Oliver, F. W. (1945): Dust-Storms in Egypt and Trans, 9th Int. Cong. Soil Sci. II.I. Their Relation to the War Period, as McDonald, E.V., F.B. Pierson, G.N. Flerchinger Noted in Maryut, 1939-45. The and L.D. McFadden (1996): Application Geographical Journal, 106(1/2): 26-49. of a process-based soil–water balance Reynolds, R.L, J. Belnap, M. Reheis, P. model to evaluate the influence of Late Lamothe and F. Luiszer (2001): Eolian Quaternary climate change on soil–water dust in Colorado Plateau soils: nutrient movement in calcic soils. Geoderma 74: inputs and recent change in source. 167–192. Proceedings of the National Academy of McFadden, L.D., E.V. McDonald, S.G. Wells, K. Sciences 98: 7123–7127. Anderson, J. Quade and S.L. Forman. Reynolds, R.L., M. Reheis, J. Yount and P. (1998): The vesicular layer and carbonate Lamothe (2006): Composition of aeolian collars of desert soils and pavements: dust in natural traps on isolated surfaces formation, age, and relation to climate of the central Mojave desert Insights to change. Geomorphology 24: 101–145. mixing, sources, and nutrient inputs. J. McLean, E.O. (1982): Soil pH and lime Arid Environments 66: 42–61. requirement. P. 199-224 in A.L. Page, Salam, A. (1967): Study of air pollution in R.H. Miller and D.R. Keeney (eds). Helwan industrial area. Report Methods of soil analysis, part 2. Chemical submitted to the high committee for and microbiological properties, 2nd planning of greater Cairo.

-106- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Salam, A. and M. Sowelim (1967): Dust deposits ecosystem and its implications for in the city of Cairo. J. Atmosph, Environ. management. Ecological Applications 8: 1: 211-220. 455–463. Schultz, L.G., (1964): Quantitative Shen, Z., S. Caquineau, J. Cao, X. Zhang, Y. interpretation of mineralogical Han, A. Gaudichet and L. Gomes (2009): composition from X-ray and chemical Mineralogical characteristics of soil dust data for the Pierre shale. U. S. Geol. from source regions in northern China. Surv., prof. Paper, No. 391-c, pp. C1-C31, Particuology xxx, 145. Washington. Whitting, L.D. (1965): X-ray diffraction Schutz, L. and M. Sebert (1987): Mineral techniques for mineral identification and aerosols and source identification. mineralogical composition. In C.A. Black Journal of Aerosol Science, 18(1): 1–10. (ed). Method of Soil Analysis. American Shachak, M. and G.M. Lovett. (1998): Society of Agronomy, Inc., Publisher. Atmospheric deposition to a desert Madison, Wisconsin, USA.

-107- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

التركيب المعدنى لرواسب رياح الخماسيـن فى أسيوط- مصر محمد األمين عبد الكريم فرج هللا*، محمود أحمد عيسى** * قسم علوم األراضى والمياه - كلية الزراعة- جامعة األزهر فرع أسيوط ** قسم الجيولوجيا – كلية العلوم - جامعة أسيوط

تهدف هذه الدراسة لمعرفة كميةة الباة ر المترسة مة رية ا الام سةي اةر اىرالةر الةرا يةة اأسةيط طتركياة المعةد ر اةة ش رةةهر لاريةةش لعةة مر 8002 ط 8002، لةةذل تةةا ااتيةة ر امةةع مطا ةةف اةةي ا فةةة الةةةارا البرايةةة إلةةر ا فةةة الةةةارا الرر ية، طتا طلف ة ديق اراية مس اته 1ا8 فر هذه اىم ك م لطش رهر لاريش اتر آاره. لظهرت ال ت ئج المتاةش ايه اعة اخات فة ت فةر كميةة الباة ر المترسة مة رية ا الام سةي مة ة ا 8002 لعة ا 8002. كمة ااتاتةت كميةةة رطاسة رية ا الام سةةي مة مط ةف لاةةر لة تع العة ا الطااةةد. طسةلات الكمية ت اى اةةر مة هةةذه الرطاس في مطا ِف الل الرر ِي لطادي ال يش الاعيةدة مة الةةارا ِ البرايةة. طالقةيا المتطسة ة لةة EC ط pH الرطاسة مة ري ا الام سي ك ت م اتلة طمتر اهة فر ك مر الدراسة. لطلح فاص العي ة ت ا لميكرطسةكط اخلكترط ةر الم سةح ل الةا المعة د المترسةاة مة رية ا الام سةي ت ة ص كامة اتله اط الررق اعيدا الةا ار ِ البراية ايث ك ت لدق فر المطا ف الرر ية ه فر المطا ف البراية لرطاس ام سي 8002 ط 8002. طاطلةة ةة ا تطلةةد كميةة ت كايةةرة مةة الااياةة ت ةاطيةةة طتاةةت ةاطيةةة الرةةكش اأالةة ا ماتاتةةة، طتطلةةد ااياةة ت مة كثيرة متا اة الركش، طكذل تطلد اعة الااياة ت كرطيةة الرةكش ا الة ا مت ط ةة، كمة لطاظةت ااياة ت يةر م تظمةة الركش فر ي ت رطاس ري ا الام سي لع مر 8002 ط 8002. لظهرت ت ئج التاص المعد ر ا ستاداا ايطد اىرعة السةي ية لرطاسة رية ا الام سةي لعة ا 8002 ل معةد الكةطارتة هط لكث ارلمع د ريط تاي مع د التاسا رات الاطت سية طالك لسيت طاخايةدطت طالم ل تيةت طاى هيةدريت طالل ر ةت مرتاةة ترتياة ت ةلي ، ثا مع د اىطليت طالتطرم لي طالةركط طاخلم يت طالا ليطكاية طالليطثيت طاىكتي طليت طالهيم تيت مطلطدة فق فر اعةة المطا ةةف. كمةة طلةةد ليلةة ل معةةد الكةةطارتة هةةط اىكثةةر رةةيط فةةر رطاسة ريةة ا الام سةةي لعةة ا 8002 تايةة معةة د التاسةةا رات الاطت سةةية طالك لسةةيت طاخايةةدطت طالم ل تيةةت طالتطرمةة لي طاى هيةةدريت مرتاةة ترتياةة ت ةلية فةةر كةةش المطا ةةف، اي مةة سلات مع د اىطليت طالا ليطكاية طاخلم يت طالرطتيش طالهيم تيت طالل ر ةت طاىكتي طليةت فةر اعة المطا ةف فقة . طخ يطلةد ه إتل ه ا لتطةي ِف هةذه المعة د ِ اةي المطا ةف المدرطسةة. طمة طلهةة ظةر ط يةة فةإ معظةا المطا ة ِف لههة تةع التركية المعد ر فر كش م مر الدراسة. طامق ر ة التركي المعةد ر لرطاسة رية ا الام سةي فةي الامسةة مطا ةف المدرطسةة اتلةح

طلطد اع اإلات ف ت الك ّمية اي ه . طا مثش هذه الدراسة تتيد فر معرفة كمية الرطاس التر تلة ف مة رية ا الام سةي سة طي طاعة اطاةةه الكيمي ئيةة مثش ماطاته طل طاع المع د المكط ة له ممة يةرثر اةر الاةطاص الماتاتةة لارالةر الةرا يةة التةر ترسةات ايهة ، كمة ل هة متيدة فر الدراس ت التر تهتا اأةش التراة ايث ل هذا الت طع فر المع د المترساة مة رية ا الام سةي طالملة فة لارالةر الةرا ية فر طادى ال يش يدش ار تعدد مطاد اىةش لارطاس التر تكط ت م ه هذه اىرالر.

-108- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

HEALTH STATUS AND RISK FACTORS OF STREET CHILDREN IN BENI-SUIEF CITY Amel Abd Elazim Mohamed*, Shokria Adly Labeeb**, Tamer Mohamed El Hafnawy*** and Asmaa Ghareeb Mohamed** *Community Health Nursing Dept., Technical Institute of Nursing, Beni-Suief University **Community Health Nursing Dept., Faculty of Nursing, Assiut University ***Public Health Dept., Faculty of Medicine, Beni-Suief University

ABSTRACT: The magnitude of street children problem is escalating, and this is related to increasing levels of poverty especially in developing countries. The aim of this study was to highlight the problem of street children in Beni-Sueif city, with emphasis on the health and social status of these children through assessing the health status of these children and identifying the most common risk factors influencing their physical, psychological, and social health. The study was conducted on 101 street children in Beni-Sueif city. Data collection tools included an interview questionnaire form, a physical assessment sheet, and a lab sheet for stool and urine analysis, hemoglobin level, and viral Hepatitis B and C markers. The results revealed that their age ranged between 7 and 16 years. The majority were current smokers, used addictive substances, and all had aggressive behavior. The most common physical problems were underweight, angular stomatitis, and teeth decay. All had positive stool analysis, 86.1% were anemic, and 24.8% had positive viral hepatitis markers. The predictors of addiction were male sex, longer duration in the street, and smoking. Age was the predictor of being hepatitis B or C positive. In conclusion, street children are mostly males who quit or never attended schools, and with untoward family circumstances. The majority are indulged in smoking, substance abuse, and violence. They suffer both physical and psychological problems. It is recommended to study the magnitude of the problem at a national level. The role of NGOs must be fostered. Screening programs are needed for these children.

BACKGROUND: street children phenomenon is an alarming and escalating worldwide problem. Street children The problem of street children is a are maltreated, imprisoned, and in some worldwide phenomenon. Many capitals and countries killed. The phenomenon of street urban centers of the world have become a haven children, and offspring of the modern urban of survival for many children in distress. The environment, represents one of humanity’s most

-109- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

complex and serious challenges (LeRoux and abandoned houses and buildings, or who move Smith, 1998). from place to place living with friends. The The problem of street children is a growing third group is children who remain in contact problem worldwide, more so in African with their family, but because of poverty, countries. The presence of large numbers of overcrowding, or sexual or physical abuse children sometimes as young as three on the within the family will spend some nights, or streets in urban areas was virtually unheard of most days on the streets. The fourth group is prior to the transition to a market economy. It is children who are in institutional care, who have now a growing problem in most African cities come from a situation of homelessness and are and towns. The problem requires urgent at risk of returning to a homeless existence. attention as it threatens the very fabric of There are very few studies available on the society. A starting point would be to get an issue of street children in Egypt, and those understanding of who these children are and the existing do not focus on the children themselves, factors that turn them into street children but instead focus on the problem and the visual (Boyden and Gibbs, 1997). indicators of its existence on a surface level, There are several factors that lead to the viewing the children as a “disease needing to be rise of the street children phenomenon in Egypt. eliminated through, in most cases, more laws The main reasons are linked to the social and and restrictions” (Bibars, 2007). Surveys of economic degradation of urban areas. About these children have found that they range in age 90% of all street children in Egypt come from from nine to eighteen years of age, although the the poor neighborhoods in urban areas. The majority are around thirteen years of age. Most immediate factors that push the children into of these children are members of the local urban the streets are abuse by family members or poor. However, some are older children that employers, neglect (due to family size, low migrate from rural areas in the hope of finding income and family breakdown), dropping out of employment. Many street children in Egypt school, and loss of job (Azer, 1993; Hope for have left families facing extreme forms of Street Children in Egypt, 2007). poverty. Many of these children do not attend According to the World Health school, do not receive health care, and are Organization (WHO, 1995), street children unprotected by adults (Zain Al-Dien, 2009). definition encompasses four different groups. The first group is children living on the street METHODS: whose immediate concerns are survival and An analytic cross-sectional study design was shelter. The second group is children who are used in carrying out this study during the year detached from their families and living in 2009-2010 in Benisuief city. The study was temporary shelters, such as hostels and

-110- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

conducted on children male or female who has personal contacts, the researcher trained two of any one of the following inclusion criteria: the specialists of nursing to assist her in  Living on the street whose immediate concerns gathering information and taking are survival and shelter. measurements of the children. Also, a Detached from families and living in laboratory technician accompanied the team for temporary shelters. the withdrawal of blood samples for analysis of the markers of hepatitis B and C viruses, and  In contact with their family, but because of poverty, overcrowding, or sexual or physical other lab tests. On the second day in the abuse within the family will spend some morning, the lab technician collected stool nights, or most days on the streets. samples. Laboratory investigations were carried out in a specialized private laboratory in Beni A snowball sampling technique was used in Sweif. The forms were filled through individual recruiting eligible children. The sample size was interviewing for each child. During the estimated to determine the prevalence of any interviews, the team offered some sweets and illness with a rate of 50% among street children, juice to children, and some children were with a 10% absolute precision and a 95% offered some clothes and small gifts through the level of confidence, using the single society of young good of the students of the proportion equation for dichotomous variables University of El-Nahda Beni-Sueif. Weight and (Schlesselman, 1982). The estimated sample size height were measured. Underweight was is 88 children. After adjustment for a dropout defined according to age- and sex-specific body rate of about 10%, the sample size was mass index (BMI) (kg/m2) as <5th percentile increased to 101. (CDC, 2000). Three tools were used to collect the Data entry and statistical analysis were necessary information. Interview questionnaire done using SPSS 14.0 statistical software form for gathering information concerning: package. Data were presented using descriptive personal data, family characteristics and details statistics in the form of frequencies and of street living. Assessment sheet includes percentages for qualitative variables, and means general examination, anthropometric and standard deviations for quantitative measurements and psychological assessment. variables. Qualitative categorical variables were Laboratory sheet for investigations of urine and compared using chi-square test. Whenever the stool analysis, Hemoglobin level and viral expected values in one or more of the cells in a Hepatitis B and C markers. Formal administra- 2x2 tables was less than 5, Fisher exact test was tive approvals were taken before the start of the used instead. To identify the independent fieldwork. Children were briefed about the predictors of various health problems, logistic study, encouraged to participate. To collect information from children through direct

-111- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

regression analysis was used. Statistical significance was considered at p <0.05

-112- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

RESULTS: According to Table 5, the most common reason for street living as reported by studied The age of children in the study sample children was to gain money (98.0%), followed ranged between 7 and 16 years, with mean by having a chance to work with friends 12.9±2.3 years. The majority (83.2%) were (41.6%). Family-related reasons were males, illiterate (41.6%), or just could read and maltreatment (36.6%), having stepparents write (38.6%). (Table 1). (34.7%), escaping from family (32.7%), and Figure 1 illustrates the sites from which having divorced parents (28.7%). children were recruited. More than half (60.4%) As regards the types of work done, they were from non-governmental organizations were mostly selling goods (74.3%), and begging (60.4%), while only 3.0% were from social (57.4%). Meanwhile, slightly more than one- defense. third of the children reported theft as a job Concerning school attendance, Table 2 (35.6%), whereas 5.0% reported sex work. shows that about one-third of the children Their income was on average 21.7±21.5 LE per (29.7%) never attended school, while only 2% day. Less than two-thirds of them were saving were currently in school. The majority of those money (61.4%), and the saved money was not who quit did it in primary phase (81.2%). As for given to anyone in about half of them (49.5%). the reasons for quitting, the most frequently (Table 6). mentioned were poverty (94.0%), seeking gain Table 7 shows that the most common means of money (75.0%), and lack of interest (67.0%). for getting food were begging and buying, As regards family characteristics, Table 3 73.3% and 56.4% respectively. As for clothing, indicates that the highest percentages of it was mainly from trash (56.4%), or begging children were of second or third birth order (52.5%). Most children reported having a (41.6%). The majority had siblings (93.1%), regular bath (82.2%). This was mostly in the with more sisters (2.4±1.6) than brothers river (34.9%) or at home (30.1%). About two (2.2±1.3). About one-third of the children had thirds (68.3%) reported bathing more than once stepmothers (33.7%), and step-siblings (33.7%), per week. and 14.9% had stepfathers. Figure 2 displays children's evaluation of Concerning the characteristics of street their present status compared to home living. living, most of the children were living alone Only about one-fourth of them (26.7%) viewed (77.2%), and for more than two months their life was worse than at home. (78.2%). The most common places for spending Concerning smoking, the majority of night were at work patron's (47.5%), or in the children were current smokers (83.2%) as street (45.5%). The time of being in the street shown in Table 8. This was mostly cigarettes was mostly at night (52.2%), and in any days of (75.0%), with a mean 16.0±11.7 cigarettes per the week (82.2%). (Table 4).

-113- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

day. The mean age at start of smoking was The most common problems were underweight 9.2±2.1 years, and it was mostly after leaving (57.4%), angular stomatitis (68.3%), and teeth home (60.7%). decay (58.4%). As for addiction and drug abuse, Table 9 As for general appearance, Table 13 shows indicates a high knowledge about bango that more than half of the children looked (91.1%), Kolla (89.1%), Hashish (74.3%), and depressed (54.5%), and almost all of them alcohol (64.4%). It also shows that about three- (96.0%) lacked personal hygiene. The total fourth of the children have ever used drugs number of problems per child ranged between 3 (74.3%), and this mostly happened in the street and 9, with meanSD 6.01.6 problems. (100.0%). Laboratory findings (Table 14) revealed Table 10 shows that all studied children had that all of the study children had positive stool aggressive behavior in the form of quarrelling analysis. The majority were anemic (86.1%), (100.0%). The most commonly mentioned tools with mean Hemoglobin 10.00.6 gm/dl. Also, used in violence were nails, stones, and razor about one-fourth of them had viral hepatitis blades, 82.2%, 68.3%, and 67.3% respectively. markers (24.8%), mostly C. On the other hand, about one-fifth of them Table 15 points to statistically significant (21.8%) committed violence against themselves. associations between smoking and child’s age The most common of this was suicidal attempts (p=0.04) and sex (p<0.001). It is evident that (43.2%) and cutting veins (18.2%). smoking was higher with older age group As regards psychological problems, Table (31.0%) and among male children (92.9%). 11 shows that all children had some kind of Similarly, Table 16 shows no statistically psychological problem (100.0%). Phobias were significant associations between smoking and the most commonly reported (83.2%), followed any of the child’s street life characteristics. by frustration (77.2%), feeling unhappy Table 17 demonstrates statistically (71.3%), and feeling like quarreling (70.3%). significant associations between drug addiction Also, 60.4% had fears from the police. and child’s age (p=0.04) and sex (p<0.001). It Physical examination of the children can be noticed that drug addiction was higher revealed that almost all of them (94.1%) had with older age group (32.0%) and among male some physical abnormal findings (Table 12). children (93.3%).

-114- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Part I: Socio-demographic characteristics of children and their families:

Social defense, 3 Institution, 8.9 Parking lot, 12.9

NGO, 60.4 Street, 14.9

Figure 1: Distribution of children in the study sample according to location (n=101)

Table 1: Socio-demographic characteristics of children in the study sample (n=101) Frequency Percent Site: Parking lot 13 12.9 Social defense 3 3.0 Institution 9 8.9 Street 15 14.9 NGO 61 60.4 Age (years): <15 74 73.3 15+ 27 26.7 Range 7.0-16.0 Mean±SD 12.9±2.3 Sex: Male 84 83.2 Female 17 16.8 Education: Illiterate 42 41.6 Read/write 39 38.6 Primary 17 16.8 Preparatory 3 3.0

-115- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 2: School attendance among children in the study sample (n=101) Frequency Percent School attendance: Never attended school 30 29.7 Entered and quit 69 68.3 Currently in school 2 2.0 Quit at: Primary 56 81.2 Preparatory 13 18.8 Currently in: Primary 1 50.0 Preparatory 1 50.0 Causes of no entry/quitting school:@ Family poverty 94 94.0 Seeking money gain 75 75.0 No interest 67 67.0 Family disorganization 47 47.0 Work to support family 38 38.0 Difficult transportation 31 31.0 Far school 29 29.0 Frequent failure 23 23.0 Maltreatment at school 20 20.0 Family will and car 22 22.0 Overcrowded school 9 9.0 Inefficient teachers 3 3.0 Disease/disability 2 2.0 Unsuitable school schedule 1 1.0 Other (father death, neglect) 10 9.9 (@) Not mutually exclusive

Table 3: Family characteristics of children in the study sample (n=101) Frequency Percent Birth order: 1 32 31.7 2-3 42 41.6 4+ 27 26.7 Have siblings 94 93.1 No. of brothers: Range 0-5 Mean±SD 2.2±1.3 No. of sisters: Range 0-6 Mean±SD 2.4±1.6 Have stepmother 34 33.7 Have stepfather 15 14.9 Have step-siblings 34 33.7 No. of stepbrothers: Range 0-5 Mean±SD 0.5±1.0 No. of stepsisters: Range 0-5 Mean±SD 0.4±0.8

-116- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Part II: Characteristics of street life:

Table 4: Characteristics of street living as reported by children in the study sample (n=101) Frequency Percent Current living status: With family 23 22.8 Alone 78 77.2 Duration in the street (months): <2 22 21.8 2+ 79 78.2 Range <1-120 Mean±SD 45.5±30.0 Spending night at:@ Work patron 48 47.5 Street 46 45.5 Family home 30 29.7 Friends 17 16.8 Relatives 4 4.0 Time of being in street: Morning 26 25.7 Afternoon 10 9.9 Night 53 52.5 Any time 12 11.9 Days of the week being in street: Weekend 8 7.9 Weekdays 10 9.9 Any time 83 82.2

Table 5: Reasons for street living as reported by children in the study sample (n=101) Reasons for living in street@ Frequency Percent To gain money 99 98.0 Job opportunity with friends 53 53.6 Parents' maltreatment 37 36.6 Stepparents 35 34.7 Escape fro family 33 32.7 Divorced parents 29 28.7 Maltreatment from others 15 14.9 Father death 13 12.9 Escape from home responsibilities 10 9.9 Looking for relatives 1 1.0 Job opportunity with relatives 1 1.0 Other (crime, father illness) 25 24.8 (@) Not mutually exclusive

-117- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 6: Work and income from street living as reported by children in the study sample (n=101) Frequency Percent Types of work done: @ Selling goods 75 74.3 Begging 58 57.4 Car washing 38 37.6 Theft 36 35.6 Janitor 34 33.7 Carrying luggage 18 17.8 Serving at homes 15 14.9 Shining shoes 9 8.9 Drug trafficking 5 5.0 Sex work 5 5.0 Destruction of properties 2 2.0 Other (baker, construction, etc.) 7 6.9 Income gained the day before (LE): <20 53 52.5 20+ 48 47.5 Range 0.0-125.0 Mean±SD 21.7±21.5 Saved money the day before (LE): No 39 38.6 Yes 62 61.4 Range 0.0-900.0 Mean±SD 42.4±133.9 The money saved given to: Nobody 50 49.5 Friend for travel 32 31.7 Friend for a project 12 11.9 Friends for food 3 3.0 Friend for room rent 1 1.0 Brothers 2 2.0 Work patron 1 1.0 (@) Not mutually exclusive

Table 7: Means of getting food, clothes, and bathing as reported by children in the study sample (n=101) Frequency Percent Means of getting food: Beg 74 73.3 Buy 57 56.4 Trash 33 32.7 Steal 15 14.9 Sex 1 1.0 Means for clothing: Trash 57 56.4 Beg 53 52.5 Buy 39 38.6 Steal 22 21.8 Sex 1 1.0 Bath 83 82.2 Bathing place: River 29 34.9 Home 25 30.1 Mosque 15 18.1 Work 6 7.2 Friends 5 6.0 Public bath 3 3.6 Frequency: Weekly 32 31.7 More than once per week 69 68.3

-118- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Part III: Psychosocial and physical health problems:

Worse, 26.7 Better, 32.7

Different, 40.6

Figure 2: Perception of current situation as reported by children in the study sample (n=101)

Table 8: Smoking habits as reported by children in the study sample (n=101) Frequency Percent Smoking: Never 17 16.8 Current 84 83.2 Type: Cigarettes 63 75.0 Hookah + cigarettes 17 20.2 Hookah + cigarettes + goza 4 4.8 Range 2.0-60.0 Mean±SD 16.0±11.7 No. of hookah fills: Range 1.0-5.0 Mean±SD 2.2±1.1 Duration (years): Range 1.0-13.0 Mean±SD 4.1±2.7 Age at start (years): <10 45 53.6 10+ 39 46.4 Range 5.0-16.0 Mean±SD 9.2±2.1 Time of start: Before leaving home 33 39.3 After leaving home 51 60.7

-119- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 9: Drug addiction & Substance abuse as reported by children in the study sample (n=101) Frequency Percent Knowledge about addictive drugs: Bango 92 91.1 Kolla 90 89.1 Hashish 75 74.3 Alcohol 65 64.4 Cough syrup 58 57.4 Tablets 51 50.5 Cocaine 41 40.6 Benzene 40 39.6 Opium 32 31.7 Other (injections, cockroach powder) 2 2.0 Range 0.0-9.0 Mean±SD 5.4±2.6 Ever used drugs: 75 74.3 Easily get drugs 35 46.7 Place of using drugs: Street 75 100.0 Coffee shop 18 24.0 Home 4 5.3 Other (workplace) 5 6.6

Table 10: Aggressive behavior as reported by children in the study sample (n=101) Frequency Percent Aggressive behavior:@ Quarreling 101 100.0 Destruction of others' properties 38 37.6 Setting fires 14 13.9 Violence commitment: One type 54 53.5 More than one type 47 46.5 Tools used: @ Nails 83 82.2 Stones 69 68.3 Razor blades 68 67.3 Knives 53 52.5 Glass 50 49.5 Chemicals 35 34.7 Others (stick) 2 2.0 Committed violence against self 22 21.8 Means: @ Suicidal attempt 19 43.2 Cut veins 8 18.2 Jumped from train 4 9.1 Jumped from height 4 9.1 Drug overdose 3 6.8 Injured self 2 4.5 Threw self in front of a car 2 4.5 Drank medication 1 2.3 Poison 1 2.3 (@) Not mutually exclusive

-120- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 11: Psychological problems as reported by children in the study sample (n=101) Psychological problem@ Frequency Percent Phobias 84 83.2 Frequent feeling of frustration 78 77.2 Feel unhappy 72 71.3 Feeling like quarreling 71 70.3 Daydreaming 64 63.4 Biting nails 61 60.4 Nightmares 54 53.5 Anorexia 50 49.5 No desire to play 37 36.6 Wetting bed 17 16.8 Other fears from: Police 61 60.4 Snakes 2 2.0 Ghosts 1 1.0 Sexual harassment 3 3.0 Adults 1 1.0 Police and adults 2 2.0 Police and sexual harassment 4 4.0 Police and dark 2 2.0 Police and beating 4 4.0 Police and death 2 2.0 Any psychological problem 101 100.0 (@) Not mutually exclusive

Part IV: Physical assessment and laboratory findings:

Table 12: Body mass index and physical exam findings among children in the study sample (n=101) Frequency Percent Body mass index (BMI): Underweight (<18.5) 58 57.4 Normal (18.5-25) 43 42.6 Eyes: Normal 20 19.8 Angular palperitis 41 40.6 Pale conjunctiva 32 31.7 Both 8 7.9 Angular stomatitis 69 68.3 Teeth: Normal 7 6.9 Decayed 59 58.4 Mottled 23 22.8 Both 12 11.9 Abnormal physical findings 95 94.1

-121- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 13: General observation findings among children in the study sample (n=101) Observation@ Frequency Percent Lack of personal hygiene 97 96.0 Active 67 66.3 Depressed 55 54.5 Cheerful 37 36.6 Serious 30 29.7 Lice in hair 28 27.7 Adequately clothed 24 23.8 Nails cut 20 19.8 Neat/tide 3 3.0 Total observed problems: <4 8 7.9 4+ 93 92.1 Range 3-9 Mean±SD 6.0±1.6 (@) Not mutually exclusive

Table 14: Laboratory findings among children in the study sample (n=101) Frequency Percent Positive stool analysis 100 99.0 Anemic 87 86.1 Hb: Range 9.0-12.0 Mean±SD 10.0±0.6 Hepatitis B +ve 4 4.0 Hepatitis C +ve 21 20.8 Any positive hepatitis 25 24.8

Part VII: Relation between smoking and addiction and various characteristics of children:

Table 15: Relation between children’s smoking and their socio-demographic characteristics Smoking X2 No Yes p-value Test No. % No. % Age (years): <15 16 94.1 58 69.0 15+ 1 5.9 26 31.0 Fisher 0.04* Sex: Male 6 35.3 78 92.9 Female 11 64.7 6 7.1 Fisher <0.001* Education: Illiterate 9 52.9 33 39.3 Educated 8 47.1 51 60.7 1.09 0.30 Birth order: 1 4 23.5 28 33.3 2-3 7 41.2 35 41.7 0.99 0.61 4+ 6 35.3 21 25.0 (*) Statistically significant at p<0.05

-122- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Table 16: Relation between children’s smoking and their street life characteristics Smoking X2 No Yes p-value Test No. % No. % Income gained the day before (LE): <20 12 70.6 41 48.8 20+ 5 29.4 43 51.2 2.69 0.10 Bath 14 82.4 69 82.1 Fisher 1.00 Perception of current status: Better than home 4 23.5 29 34.5 Different from home 8 47.1 33 39.3 0.79 0.67 Worse than home 5 29.4 22 26.2

Table 17: Relation between children’s addiction and their socio-demographic characteristics Drug addict X2 No Yes p-value Test No. % No. % Age (years): <15 23 88.5 51 68.0 15+ 3 11.5 24 32.0 4.13 0.04* Sex: Male 14 53.8 70 93.3 Female 12 46.2 5 6.7 Fisher <0.001* Education: Illiterate 13 50.0 29 38.7 Educated 13 50.0 46 61.3 1.02 0.31 Birth order: 1 10 38.5 22 29.3 2-3 9 34.6 33 44.0 0.92 0.63 4+ 7 26.9 20 26.7 (*) Statistically significant at p<0.05

DISCUSSION: girls ranges from only 3% to 29%, which is consistent with the present study. A similar age The present study involved a sample of range was reported in Philippines (Shanahan, children in the age range between 7 and 16 2003), and in Nigeria (Oyeniyi, 2010). The years, with the majority being males. These are disparity in age difference between of children the typical age and sex characteristics of street in developed countries and developing countries children, although there are variations from may be due to the available social resources in place to place. In congruence with this, Schecter developed countries (Boakye-Boaten, 2006). and Villamizar (2000) mentioned that in Moreover, the mean of 12.9 years of developing countries, children under the age of children in the present study is almost equal to 5 years are rarely found on the street without that reported by the CAPMAS (2009) in Cairo the supervision of a family member, while street children, which is around 13 years. It also starting at the age of eight years, children live reported a proportion of 85% boys, which is completely on their own in the street. Also, most quite close to the proportion of boys (83.2%) in street children are boys, and the proportion of

-123- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

the present study. This sex difference has been survey, where the poor economic conditions at attributed to that girls are normally involved in home constituted the main reason for quitting activities which keep them at a distance from school, followed by parents' unwillingness to streets, such as domestic work in private houses. send them to school also due to economic On the other hand, boys are more likely to be reasons, and then their lack of interest in involved in street activities as washing cars, education (CAPMAS, 2009). shoe-shining, and peddling goods such as paper According to the present study findings, tissue boxes to drivers at traffic lights Hatløy only about one-fourth of the street children and Huser, 2005). Additionally, Abdelgalil et al. viewed their life was worse than at home, while (2004) suggested that this gender imbalance about one third viewed it better. Only less than could be due to the fact that in developing one-tenth of them hoped to go back home. The countries girls are taught to cope with poverty finding is in congruence with the street children while staying at home. Another reason given by study in Cairo and Alexandria (UNODCCP, Aptekar (2000) was that boys are expected to 2009), which revealed that only 12% of contribute to the household income and thus, respondents identified returning to their many go to the street to do so. families as a first need. However, it is in The majority of street children in the disagreement with le Roux and Smith (1998) present study were illiterate, or just could read who showed that most street children aspire to and write. Only 2% of them were currently in return home, provided that the familial factors school. The findings are in line with the that drove them away change. The discrepancy CAPMAS (2009) report, which indicated that with the present study could be related to the most street children in Cairo are deprived of lower socio-economic levels of families, which basic education. However, this report rates of makes children feel better in the street. currently attending school (43%) is much As for their future hopes, as expressed higher, compared to the present study. The by children in the present study, they difference could be due to the setting, as the were mostly getting married, and getting a present study was carried out in a city of Upper good job. The findings indicate that these Egypt where school attendance is expected to be children have adapted themselves to street lower, compared to the Capital. living, and hope just to improve their conditions According to the present study findings, therein. In congruence with this, West (2003) economic factors as poverty and seeking gain of clarified that while some street children express money were the most common reasons concern about their long-term futures, most see mentioned for quitting school. These were this being largely beyond their control. Thus, followed by lack of interest in education. These many street children live for the moment, day- results are in agreement with those of the Cairo to-day.

-124- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Almost all children in the present study had those of Salem and Abd El-Latif (2002) in friends who were mostly living in the street, 12 Alexandria, where all studied street children years or older, and males more than females. were found to be were addicted to more than Having friends is essential for street children for one substance, including cigarettes smoking. On both physical and psychological support. the same line, the UNODCCP (2009) study in Moreover, due to hardship of street living, these Cairo and Alexandria found that 66% of the friends would be better strong and able to help. sample of street children consume various This might explain why children prefer older substances or drugs on a habitual basis, and friends of male gender. In line with this, Garrett 70% are cigarette smokers. et al. (2008) reported that nearly all participants Concerning the psychological problems, the of street children mentioned the importance of present study findings indicated that all developing very deep bonds with friends or children had experienced some sort of “street family” who provide material and psychological problem. The most common were emotional support, as well as a sense of phobias, frustration, feeling unhappy, feeling protection, belonging, and identity. Moreover, le like quarreling, and fears from the police. Also, Roux and Smith (1998) mentioned that when most of them have the feeling that they are street children band together, they represent an inferior to their peers. These problems reflect exceptional companionship system, which the constant feeling of insecurity in which these replaces the family as a source of emotional and children live all the time. They are afraid of economic support. The group offers protection, everything and of every person. They lack support, friendship, and solidarity. confidence in all society, and this fosters The development of risky health habits such aggression among them as a defensive as smoking and drug abuse is a special concern mechanism against the unknown. In congruence in the problem of street children. In the present with these present study findings, le Roux and study, the majority of children were current Smith (1998) reported that street children fear smokers. The mean age at start of smoking was being harmed, incapacitated, arrested, and about nine years, and it was mostly after leaving getting sick. They also are concerned about home. Moreover, most of them had high loneliness and being unloved. They desire knowledge about various addicting substances, respect and tend to see themselves as nice people and about three-fourth have ever used drugs. who behave. These high rates are quite expected given the On physical assessment, almost all of the feelings of freedom in the street with no street children of the present study had at least supervision, the availability of money and one abnormal physical finding. The most addictive substances, as well as the peer common problems were underweight, angular pressure. The findings are in agreement with stomatitis, and teeth decay, which affected

-125- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

about two-thirds of them each. These problems personal hygiene, and more than half looked are a reflection of the malnutrition and poor depressed. Meanwhile, more than two-thirds lifestyle, and confirm the complaints and looked active, and more than one third looked symptoms previously reported by these cheerful. These findings indicate that despite the children. Although these children earn money poor living conditions, many of the street and many of them can save, they still suffer mal- children are coping, and try to adapt for their or under-nutrition. This might be due to the new lifestyle. On the same line, Ayuku et al. previously explained reason whereby children (2004) highlighted that street children tend to be spend more money on buying tobacco and highly resilient displaying a high degree of addictive substances. Another reason could be adaptability and flexibility in the face of the lack of knowledge about proper nutrition. adversity and, because of their special In agreement with the foregoing, Thapa et psychological characteristics, remaining al. (2009) explained that street children are remarkably well adjusted as individuals. They often underweight despite bigger earnings also have vigor and overt motor activity in a because of the acquisition of poor eating habits routine and constant struggle to get food. and food lacking the essential nutrients. They Furthermore, Carter et al. (2001) described how found that 43.8% of the street children in the street children must develop a variety of coping sample were underweight. Similarly high rates strategies to avoid the theft and beatings. The of underweight and malnutrition were reported low personal hygiene manifestations have also among street children in Kenya (Ayaya and been previously reported in many street Esamai, 2001), Ghana (Pemberton, 2007), and children studies (Christophe, 2007; Pemberton, Manila (DOST, 2007). 2007; Thapa et al., 2009). Conversely, West (2003) claimed that street In further confirmation of the present study children may be better nourished than their symptoms and signs, laboratory investigations peers who live in poverty in rural areas. This were performed. The findings demonstrated offers a reason why some children move from that all of the study children had positive stool their homes into conditions that might be an analysis for parasites, and the majority were improvement only in some respects. However, anemic (86.1%). Moreover, about one-fourth of for some children, the street may be better than them had viral positive viral hepatitis markers home or the local institutional care, but this (24.8%). The very high prevalence of parasitic does not mean that either is desirable; infestations is quite expected given the low alternative or improved care is necessary in hygienic conditions. It has its impact on the such cases. nutritional status, and may contribute to the Apart from the signs of malnutrition, almost very high prevalence of anemia. Meanwhile, the all children in the present study sample lacked high prevalence of viral hepatitis infections can

-126- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

be attributed to promiscuous sexual relations, as street children are at risk of encountering or well as addiction in the form of injectables. interfering in violence, sexual abuse, substance The very high rate of parasitic infestation addiction and of acquiring infectious diseases revealed in the present study is in agreement including viral hepatitis. However, the with that reported among street children in prevalence of viral hepatitis markers in the Ghana (Pemberton, 2007) where 92% of the present study is even higher than that reported participants were having such infestations. On in the American literature, where it varies from the contrary, much lower rates of stool test 12 to 21% among street children (Ogilvie et al., parasite infestations were demonstrated in 1999; Beech et al., 2002). The higher prevalence Nepal in the study of Christophe (2007), which in the present study might be attributed to that reported a rate of 18%, and the study of Thapa the disease is endemic in Egypt, with very high et al. (2009), which reported that only 8.4% of prevalence rates. the respondents had worm infestations. The Analysis of the factors associated with differences among various studies could be various risks and problems revealed a number attributed to differences in the endemicity of of significant relations. The younger age various parasites in different settings. children were more prone to malnutrition and Similar to the present study findings underweight and other physical problems, regarding anemia and malnutrition, Singh et al. which also increased by lower income. On the (2008) reported that the most common other hand older age children are more prone to morbidity problems among street children in smoking, addiction, violence, sex, and viral India are anemia, malnutrition, and dental hepatitis infections. Also, the smoking, caries. Also, Patel (2006) reported malnutrition addiction, and violence problems are all inter- and anemia in majority of the street children as related. These findings are quite plausible since a result of poor diet, poor hygienic condition younger children are "newcomers" to street life, and ignorance of these children regarding with less survival experience. On the other nutritive food. The study in Alexandria has even hand, older ones experience the adolescence risk reported higher prevalence rate of underweight, behaviors of smoking, addiction, and but a slightly lower rate (78%) of anemia promiscuous sex, which expose them to sexually (Salem and Abd El-Latif, 2002). Close figures of transmitted infections as viral hepatitis. anemia were also reported in Cairo by Study Limitations: Eltahalwy et al. (2006) and in Ghana by  The first problem faced the researcher was the Pemberton (2007). difficulty in interviewing the children and Furthermore, and in line with the present finding them. study finding pertaining to viral hepatitis  The second problem, children were reluctant infections, Türkmen et al. (2004) reported that to participate' when privacy was secured in a

-127- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

place where they could talk freely, the hepatitis infection. Therefore, these children children were more willing to talk about their need both physical and psycho-social support. problems which they considered very personal, and thus should not be talked The main recommendations are about. Gaining the confidence of children and summarized as the following: ensuring total confidentiality helped the  Programs addressing the needs of street investigator to collect data. children constitute a form of rehabilitation  The task was more difficult because blood which is considered too little, too late and not samples had to be taken. To deal with this inefficient tool for proper prevention. So, problem, the researcher was accompanied by efforts should be addressed at earlier level of some delegates from the Assembly of interventions to the target population homeless children to convince them and knowing the risk factors. clarify the aim of the study to them. They also  Efforts should be addressed to the families, gave them a cash bonus and were encouraged especially those with large numbers of to bring other children to the headquarters of children and living in slum areas to help them the Assembly. to overcome their economic and social problems that push their children to the CONCLUSION AND RECOMMENDATIONS: street. Street children are among the most  There is a need for concerted efforts among vulnerable social group in any society. The local NGOs to implement educational study findings lead to the conclusion that street programs that focus on the health care needs children are mostly males who quit or never of street children. attended schools. The untoward family circumstances such as poverty, illiteracy, REFERENCES: quarreling, separation, divorce, and stepparents Abdelgalil S., Gurgel R.G., Theobald S., and push these children to street life. They are Cuevas L.E. (2004): Household and involved in theft and commercial sex, and the family characteristics of street children in majority indulges in smoking and substance Aracaju, Brazil. Arch Dis Child; 89: 817– abuse as well as violence. They suffer both 820. physical and psycho-social health problems. The Aptekar L. (2000): New directions on most common physical problems are related to adolescents research: "Developing a cross poor nutrition and parasitic and microbial - cultural methodology for working with infections, where about one-fourth are positive street children". Paper presented at the for viral hepatitis markers. The predictors of European Association for Research on drug abuse are male sex, longer duration in the Adolescence. University of Jena, June 2, street, and smoking, while age is the predictor of 2000.

-128- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ayaya S.O., and Esamai F.O. (2001): Health menacpi.org/CPI/uploads2009/Activities/ Problems of the Street Children in Street_Children _Survey_Cairo_2009.pdf Eldoret, Kenya. East Africa Med Carter J.D., Joyce P.R., Mulder R.T., Luty S.E. Journal.; 78(12): 624-9. (2001): The Contribution of Temperam- Ayuku D.O., Devries M.W., Mengech H.N.K.A., ent, Childhood Neglect, and Abuse to the and Kaplan C.D. (2004): Temperament Development of Personality Dysfunction: characteristics of street and non-street A Comparison of three Models. J of children in Eldoret, Kenya. Afr Health Personality Disorders.;15(2):123–135. Sci.; 4(1): 24–30. Christophe J.R. (2007): Study, approaches and Azer A. (1993): Situation Analysis of comments on the daily life of the street- Disadvantaged Children in Egypt, based children of the Nepalese capital, UNICEF, Cairo. Kathmandu: CPCS. Beech B.M., Myers L., and Beech D.J. (2002): DOST (2007): [homepage on the internet]. Hepatitis B and C infections among [Cited on 14th May, 2008]. The homeless adolescents. Fam Community Nutritional Status of Street children: Health; 25:28-36. Why some are nutritionally well-off Bibars I. (2007): "Street Children in Egypt: or worst off? Available from: From the Home to the Street to http://www1.fnri.dost.gov.ph/htm/streetc Inappropriate Corrective Institutions." hildren.htm. Environment and Urbanization; 10(1): Eltahalwy E.M., Schawki M.M., Ghobashi 201. (1998). M.M., AUC Social Research Center. Boakye-Boaten A. (2006): An Examination of (2006): An Evaluation of the Health The Phenomenon of Street Children In condition of street children in Cairo. Selected Communities In Accra (Ghana). Public Health and Human rights, APHA A dissertation presented to the faculty of 134th Annual Meeting and Exposition. the College of Education of Ohio Garrett S.B., Higa D.H., Phares M.M., Peterson University In partial fulfillment of the P.L., Wells E.A., and Baer J.S. (2008): requirements for the degree Doctor of Homeless Youths' Perceptions of Services Philosophy November. and Transitions to Stable Housing. Eval Boyden J., and Gibbs S. (1997): Children and Program Plann. November; 31(4): 436– War: Geneva: Understanding Psych- 444. ological Distress in Cambodia, UN. Hatløy A., and Huser A. (2005): Identification of CAPMAS (2009): A profile of Cairo street street children. Characteristics of street children: initial results. Accessed on children in Bamako and Accra. Fafo- 23/4/2010. Available at: http://www. report 474, Oslo: Fafo.

-129- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Hope for Street Children in Egypt (2007): Singh D., Sareen N., Ojha A., and Sareen D. United Nations Office on Drugs and (2008): Street Children of Udaipur: Crime. Accessed on 9/4/2010 at < Demographic Profile and Future http://www.unodc.org/unodc/newsletter_ Prospects. Stud Tribes Tribals, 6(2): 135- 2001 -12-01_1_page 007.html>. 139. Le Roux J., and Smith C.S. (1998): Causes and Thapa K., Ghatane S., and Rimal S.P. (2009): Characteristics of the Street Child Health problems among the street Phenomenon: A Global Perspective. children of Dharan municipality. Adolescence; 33 (Issue 131). Kathmandu University Medical Journal; Ogilvie E.L., Veit F., Crofts N., and Thompson 7( 3): Issue 27, 272-279. S.C. (1999): Hepatitis infection among Türkmen M., Okyay P., Ata Ö., and adolescents resident in Melbourne Okuyanoðlu S. (2004): A descriptive Juvenile Justice Centre: risk factors and study on street children living in a challenges. J Adolesc Health; 25:46-51. southern city of Turkey. The Turkish Oyeniyi A.J. (2010): Attitudes of street children Journal of Pediatrics; 46: 131-136. to the network of support for them in UNODCCP-United Nations Office for Drug nigeria. Ife Psychologia;. Control and Crime Prevention, (2009): Patel S.K. (2006): Child labour in Agrabati Rapid Situation Assessment of street making in slum area of Nagpur. In children in Cairo and Alexandria, p. Abstracts of the Souvenir Published it in 16-17 [Online], available at 43rd National Conference of IAP. http://www.unodc.org/pdf/youthnet/egypt Pemberton S. (2007): Saving the street Children _street_ children_report.pdf Kiwanis Magazine. Indianapolis: Kiwanis West A. (2003): At the Margins: Street Children International, pp. 1-6. in Asia and the Pacific. Asian Salem E.M., and Abd El-Latif F. (2002): Development Bank. Sociodemographic characteristics of WHO (1995): Papers on Violence and Health street children in Alexandria. Eastern Pathways and Protection from interper- Mediterranean Health Journal; 8(1). sonal Violence. Copenhagen: WHO Schecter K., and Villamizar (2000): Who Cares? contribution to World Summit for Social An Urban Age Magazine Special Report Development. on Street Children. Urban Age Magazine. Zain Al-Dien M.M. (2009): Education for Street Shanahan P. (2003): Streets versus elites: Children in Egypt: The Role of Hope Tensions, trade-offs and treaties with Village Society. Journal of Contemporary street children in Accra, Ghana. Children Issues in Education; 4(1): 37-56. Youth and Environments, 13(1).

-130- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

الحالة الصحية والعوامل الخطرة المؤثرة على أطفال الشوارع في مدينة بنى سويف أمل عبد العظيم محمد مشهور*، شكرية عدلي لبيب**، تامر محمد الحفناوى***، أسماء غريب محمد**** * ماجستير تمريض صحة مجتمع – كلية التمريض - جامعة أسيوط **أستاذ تمريض صحة المجتمع - كلية التمريض - جامعة أسيوط *** أستاذ مساعد الصحة العامة - كلية الطب - جامعة بنى سويف **** مدرس صحة المجتمع - كلية التمريض - جامعة أسيوط

أطفال الشوارع هم المجموعة االجتماعية األكثر ضعفا في أي مجتمع. وحجم هذه المشكلة متصاعد، ويرتبط هذا التصاعد بزيادة مستويات الفقر وخاصة في البلدان النامية. كان الهدف من هذه الدراسة تسليط الضوء على مشكلة أطفال الشوارع فيي مدينيية بنييي سييويف، مييع التركيييز علييى الوضييع الصييحي واالجتميياعي لهيييالء األطفييال ميين خييالل تقييييم الحاليية الصييحية لهيييالء األطفال، وتحديد عوامل الخطورة األكثر شيوعا ، والتي تيثر على الصحة الجسدية والنفسية واالجتماعية لديهم. شملت الدراسة عينة مكونة مين 101 مين أطفيال الشيوارع. تيم اسيتخدام األدوات التاليية لجميع البيانيات: اسيتمارة مقابلية، ئاةمية تقيييم الحالية البدنية، وئاةمة مختبر لتحليل البراز والبول، ومستوى الهيموجلوبين بالدم، وفحوص االلتهاب الكبدي الفيروسي )بي وسي(.

أسفرت نتاةج الدراسة عما يلي: 1- تراوح سن األطفال بين 7 و11 سنة. 2- أغلبية األطفال كانوا من المدخنين الحاليين 3- كانت المشاكل الجسمانية األكثر شيوعا لديهم هي نقص الوزن، وأعراض سوء التغذية، وتسوس األسنان. 4- العينة كلها إيجابية في تحلييل البيراز، بينميا 1161٪ يعيانون مين فقير اليدم، 2461٪ كيانوا إيجيابيين لفحوصيات االلتهياب الكبدي الفيروسي. 5- المستقل الوحيد ذا تبين أن العوامل التنبيية المستقلة ذات الداللة اإلحصاةية لإلدمان هي جنس الطفيل )اليذكورة(، والميدة األطول في الشارع، والتدخين. 1- كان السن هو العامل التنبيي ذا الداللة اإلحصاةية لإلصابة بااللتهاب الكبدي الفيروسي. الخالصيية وجييدت الدراسيية أن أطفييال الشييوارع هييم فييي الغالييب ميين الييذكور الييذين تسييربوا ميين المييدارس أو لييم يلتحقييوا ئييط بالتعليم، مع الظروف العاةلية غير المواتية. وأغلبهم منغمسون في التدخين، وتعاطي المخدرات والعنف. توصى الدراسة بعمل تقييم لحجم المشكلة على المستوى الوطني. كميا يجيب تعزييز دور المنظميات غيير الحكوميية. أيضيا هناك حاجة لعمل برامج للفحص الطبي ألطفال الشوارع.

-131- Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

AUCES

THE BIOLOGY OF EGYPTIAN WOODY PERENNIALS 4. Dalbergia sissoo Roxb. ex DC. Kamal H. Shaltout and Amr E. Keshta

Botany Department, Faculty of Science, Tanta University, Tanta, Egypt

REVIEW ARTICLE:

ABSTRACT: The present article is the fourth in a series of review articles deal with the biology of Egyptian woody perennials. Available literatures dealt with the biology of the invasive tree Dalbergia sissoo Roxb. ex DC. in Nile Delta was reviewed. The area of natural distribution is the foothills of the Himalayas from eastern Afghanistan through Pakistan and India to Nepal. Dalbergia sissoo was introduced to Egypt by Ibrahim Basha in the age of Mohamed Ali (1805-1848) for ornamental and timber purposes. This review includes the nomenclature and taxonomy of the plant, its distribution, morphology, habitat and environment, phytomass production, control measures, propagation and management, pests and diseases, economic uses and ecological importance. Ecotypic variability among its local populations needs further studies, particularly the physiological and genetical adaptations along the prevailing environmental conditions.

INTRODUCTION: 2000). El-Sheikh (1989, 1996) recorded Dalbergia sissoo along the terraces of highway Dalbergia sissoo was introduced to Egypt by roads and canals in Nile Delta, but not along the Ibrahim Basha in the age of Mohamed Ali drain banks. Slima (2006) recorded it along the (1805-1848) for ornamental and timber canal terraces and slopes in Nile Delta. This purposes (Othman 1939). El-Hadidi and Boulos plant reproduces by seeds and root suckers (1988) described it as a deciduous tree, up to 25 (Neelu et al., 2002, James 2002). The strategy of m high, bark grayish-brown, leaves pale green, reproduction by seeds and root suckers and the imparipinnate, leaflet 3-5, pale yellow flowers rapid growth rate lead to increase the plant appear in spring, The area of natural dissemination into new regions. Now, this tree distribution of this plant is the foothills of the has become a weed and spread rapidly in other Himalayas from eastern Afghanistan through terrestrial habitats of the Nile Delta such as Pakistan and India to Nepal (Appanah et al., roadsides, railways and wastelands (Fahmy

- 131 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

2007 & Keshta 2010). This may cause characters, leaf and leaflet size and shape, obstruction and difficulties in some habitats of branching and leaf density and stem surface this region. characteristics. The genetic variability of The present article is the fourth in a series these varieties was confirmed through DNA of review articles dealing with the biology of finger printing. Dalbergia sissoo has the Egyptian woody perennials (see Shaltout 2003, following taxonomic hierarchy (Wunderlin & Shaltout et al., 2006, Shaltout and Slima 2007). Hansen 2002): Kingdom: Plantae (plants), It aims to review the available literatures dealt Subkingdom: Tracheobionta (vascular plants), the its biology in Nile Delta, assessing its Super division: Spermatophyta (seed plants), distribution in different habitats and evaluate Division: Magnoliophyta (flowering plants), its ability to adapt with different environmental Class: Magnoliopsida (dicotyledons), Subclass: conditions. Such type of review articles may Rosidae, Order: Fabales, Family: Fabaceae, focus the attention of the Egyptian plant Genus: Dalbergia L. f. and Species: Dalbergia biologists to fill the gaps of information about sissoo Roxb. ex DC. the local populations of the woody perennials in the Egyptian flora and to innovate the earlier DISTRIBUTION: studies. The area of natural distribution is the foothills of the Himalayas from eastern NOMENCLATURE AND TAXONOMY: Afghanistan through Pakistan and India to Generic name Dalbergia honours the Nepal. It is a primary coloniser of new alluvial Swedish brothers Nils and Carl Dalberg, who soils along riverbanks and forms forest, either th lived in the 18 century. The former was a pure or mixed with other species. It often occurs botanist and the latter explored Surinam in association with Acacia catechu (Appanah et (Lanzara & Pizetti 1978). It is a pan tropical al., 2000). Native range includes Pakistan, genus with 100 species distributed in different Oman, Bhutan, India, Nepal and Myanmar. parts of tropical Asia, America and Australia Uncertain native status includes Iran, (Thothathn 1987). Its vernacular names include Afghanistan, Bangladesh and Malaysia; while sarsoo, shisham, sissoo, sisu, tahli and Indian known introduced range includes United States Rosewood (Bekele-Tesemma et al., 1993). Its (Florida and Arizona), Puerto Rico, Costa Rica, synonym is Amerimnon sissoo (Roxb. ex DC.) Cyprus, Benin, Cameroon, Ethiopia, Gabon, Kuntze. Dalbergia is a large genus of small to Ghana, Kenya, Mauritius, Nigeria, Senegal, medium-size trees, shrubs and lianas (family South Africa, Sudan, Tanzania, Togo, Fabaceae, subfamily Faboideae). Javaid et al. Zimbabwe, Australia, Indonesia, Thailand, Sri (2004) reported nine varieties of Dalbergia Lanka, Taiwan and Palestine (Duke 1983, sissoo that identified on the basis of physical Pallewatta et al., 2003, Wu et al., 2003, GBIF appearance of the plant, branching pattern, pod 2007, USDA-NRCS 2007, AWC undated). It is

- 132 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

increasingly planted as a street tree in southern 2.5-3.7 cm long, inconspicuous, white to dull Florida, and now it is becoming an invasive yellow and fragrant. In addition, Bangarwa species (Parrotta 1989). (1996) reported that flowers bisexual, small, MORPHOLOGY: pale yellow, in 10-15 cm long panicles that are conspicuously hairy when young. Pain and Roy 1-Macromorphology: (1981) reported that flowers are whitish to pink, El-Hadidi and Boulos (1988) described 1 cm long and in dense clusters, 5-10 cm in Dalbergia sissoo as a deciduous tree, up to 25 m length; while MacDicken (1994) reported that high, bark grayish-brown, leaves pale green, flowers 5-8 mm long, pale white to dull yellow, imparipinnate, with zigzag rachis; leaflet 3-5, racemes 2.5-3.7 cm long in short axillary alternate, broadly ovate to sub-orbicular, entire, panicles. acuminate, terminal leaflet long stalked; pale El-Hadidi & Boulos (1988) described pod as yellow flowers appear in spring, sub-sessile, in thin, yellowish-brown when ripe, 1-4 seeded. short axillary panicles; calyx bell shaped, with 5 The pods of sissoo when ripe contain 1-3 seeds, short teeth; petals much longer than the calyx; indehiscent, reniform flat, light brown, with ovary hairy, style glabrous, strap shaped delicate papery testa (Zabala 1990). Fruit is 5-7 (Fig. 1). In a recent study by Keshta (2010), he ×0.08×1.2 cm, strap-shaped, pale brown, mostly reported the prescence of some evergreen 1-seeded, less often 3-seeded (Alam et al., 2001). Dalbergia sissoo population in Minufiya and Pods are 4.5-10×0.07×1.5 cm, linear-oblong, Kalyobia governorates. Duke (1983) in West indehiscent, stipulate, glabrous, apex acute to Lafayette (Indiana-USA) described this plant as obtuse, conspicuously reticulated against the a deciduous tree with an open spreading crown seeds, usually 1-4 seeded (PIER 2006). The that reaches a height between 15-35 meters. The length of pods varies from 5-10.5 cm according primary root long, tapering, lateral roots to the number of seeds. Pods with 1-4 seeded numerous distributed down main roots; nodule were observed, amongst these 2-seeded pods are present and has long superficial roots, which common, followed by 1-seeded and 3-seeded sends up to suckers where injured (Troup 1921). (Fig. 1); while 4-seeded pods are very rare It has a long taproot and an extensive lateral (Kanak & Sahai 1994). Fruits are indehiscent, root system, often at the soil surface and 5-7.5 cm long and 8-13 mm wide (ICRAF producing suckers (PIER 2006). The leaves are undated), rounded with minute points, pale alternately arranged, compound and oddly brown in color (PIER 2006), and persistent on pinnate (Gilman & Watson 1993), with 3-5 the tree (Gilman & Watson 1993). Seeds (8-10)× glabrous, leathery leaflets, elliptical to ovate, (4-5.5) mm, brown to brownish black, reniform tapering to a point and 2.5-3.6 cm in diameter and compressed (Tewari 1994). The seeds are 6- (ICRAF undated). PIER (2006) reported that 9 mm long and number of seeds per kg. is 44000 flowers are sessile, arranged in axillary panicles, in Africa (Leloup 1956), 53000 in Bangladesh

- 133 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

(Banik 1992) and 55000 kg in Arunachal Pradesh (Beniwal & Singh 1989).

Fig. 1: Dalbergia sissoo. A: leaves, B: flowers, C: fruits and D: Dalbergia sissoo tree in Abo Al-Akhdar canal at Zagazig

2-Micromorphology: pericycle fibers and vascular tissue. Secondary xylem is formed from xylem vessels and fibers A-Stem: and xylem phloem ray parenchyma present The epidermis of stem has a single layer of between main vascular bundles which are radial cells which are tubular in cross section. They as the stem is old dicot vine stem. Cambium is are closely packed without intercellular spaces. present between xylem and phloem tissues. In Under epidermis there are patches of lamellar the centre of the stem, pith cells present which collenchyma cells. Cortex consists of 4-6 layers are spherical in shape with intercellular spaces. of spherical parenchyma cells with intercellular spaces (Fig. 2). Also, there are patches of extra- B-Root: xylary pericycle fibers above the main vascular Cortex of old root is wider than that in the bundle. Phloem tissues are present between stem, and endodermis is destroyed due to the

- 134 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

presence of secondary xylem tissues (Fig. 3). The stele is made up of radially arranged strands of xylem and phloem and the pith is E-Evergreen leaf: very compressed in the centre. Arms of primary Leaf of the plant is bifacial with flat upper xylem are present in the centre of the root and and lower epidermis (Fig. 6). The leaf is made very compressed due to formation of secondary up of upper and lower epidermis with the xylem. mesophyll in the middle which is made of parenchyma cells. There are thin layer of cuticle C-Deciduous leaf: covered the epidermis layer. Epidermis is made Leaf of the plant is bifacial with flat upper up of one layer of closely backed tubular cells and lower epidermis (Fig. 4). The leaf is made with thin walls. In the lower epidermis, there up of upper and lower epidermis with the are stomata which are present in grooves. The mesophyll in the middle which is made of vascular bundle is present in the midrib region parenchyma cell. There are thin layer of cuticle in rows in an inverted form. In the midrib covered the epidermis layer. Epidermis is made region and below upper and lower epidermis, up of one layer of closely backed tubular cells there are cartilaginous collenchyma cells. There with thin walls. In the lower epidermis, there are circle ring of fibers in the midrib region that are stomata which are present in grooves. In the surround the vascular bundle. midrib region, the vascular bundle is present in rows in an inverted form. In the midrib region F-Evergreen leaf petiole: and below upper and lower epidermis, there are There are thin layer of cuticle covered the cartilaginous collenchyma cells. epidermis layer (Fig. 7). Epidermis is made up of one layer of closely backed tubular cells with D-Deciduous leaf petiole: thin walls. Cortex is made up of 3-5 rows of There are thin layer of cuticle covered the spherical parenchyma cells. Vascular bundle is epidermis layer (Fig. 5). Epidermis is made up in the form of circle and made up of phloem of one layer of closely backed tubular cells with followed by cambium tissues and xylem is thin walls. Cortex is made up of 3-5 rows of arranged in rows separated by secondary fibers. spherical parenchyma cells. Vascular bundle is Outside the vascular bundle there are extra- in the form of circle and made up of phloem xylary fibers in the form of patches upon the followed by cambium tissues and xylem is main vascular bundle. Pith in the centre is made arranged in rows separated by secondary fibers. of spherical parenchyma cells with intercellular Outside the vascular bundle, there are patches spaces. of cartilaginous collenchyma cells. Pith in the centre is made of spherical parenchyma cells G-Pod punches petiole: with intercellular spaces.

- 135 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Epidermis is made up of one layer of closely H-Fruit petiole: backed tubular cells (Fig. 8). There are 2-3 Epidermis is made up of one layer of closely layers of cork above the epidermis. Cortex is backed tubular cells. Thin layer of cuticle cover made up of 4-6 rows of spherical parenchyma the epidermis (Fig. 9). Cortex is made up of 4-5 cells. The first two rows of cortex forming ring rows of spherical parenchyma cells. Six-seven of cartilaginous collenchyma cells. Vascular vascular bundles in the form of circle and made bundle is in the form of circle and made up of up of phloem followed by cambium tissues and phloem followed by cambium tissues, and xylem xylem is arranged in rows. Outside the vascular is arranged in rows separated by secondary bundle, there are patches of extra-xylary fibers. Outside the vascular bundle there are pericycle fibers upon each vascular bundle. Pith ring of extra-xylary pericycle fibers. Pith in the in the centre is made of spherical parenchyma centre is made of spherical parenchyma cells cells with intercellular spaces. with intercellular spaces.

Fig. 2: Transverse section through the old stem of Dalbergia sissoo Roxb. ex DC

- 136 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Fig. 3: Transverse section through the old root of Dalbergia sissoo Roxb. ex DC

Fig. 4: Transverse section through the deciduous leaf of Dalbergia sissoo Roxb. ex DC

Fig. 5: Transverse section through the deciduous leaf petiole

- 137 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

of Dalbergia sissoo Roxb. ex DC

Fig. 6: Transverse section through the evergreen leaf of Dalbergia sissoo Roxb. ex DC

Fig. 7: Transverse section of the evergreen leaf petiole of Dalbergia sissoo Roxb. ex DC

Fig. 8: Transverse section through the pod punches petiole of Dalbergia sissoo Roxb. ex DC

- 138 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Fig. 9: Transverse section of the fruit petiole of Dalbergia sissoo Roxb. ex DC

HABITAT AND ENVIRONMENT: (Black & Meerow 1993). Dalbergia sissoo is Distribution of Dalbergia sissoo ranges from found in tropical to sub-tropical climates in sea level to >1500 m above sea level, it can stand natural and planted forests, mainly along forest temperatures from below freezing to nearly margins near streams and rivers, hammocks, 50°C. It is adapted to savanna woodlands where canopy gaps, agricultural areas, disturbed sites annual rainfall is 7-20 mm (NAS 1979 & Sheikh and roadsides (Langeland & Stocker 2001, 1989). The trees are distributed from Bhutan in Duke 1983 and Sharma et al., 2000). It grows the East to Afghanistan in the west. In Nepal it best in porous well-drained soils like sands, is distributed from the terai up to 1400 m sandy loams, gravels, and alluvial soils; but does (Napier & Robbins 1989). Sissoo is native to the poorly in heavy clay and waterlogged soils foothills of the Himalayas of India, Pakistan and (Sharma et al., 2000). In India, abundant Nepal. It is expected to tolerate annual moisture and lack of competition is the key to its precipitation of 6 to 40 mm, annual temperature successful regeneration; it is therefore found in of 21 to 28°C, and pH of 6-8. It prefers well riverine environments where sunlight and drained, alluvial soils near rivers and streams. moisture are plentiful, associated with Pinus In its native range, the temperature averages roxburghii, Acacia catechu and Shorea robusta 12-22oC, and annual rainfall 500-2000 mm. It is (Hocking 1993). distributed in a monsoonal pattern with Dalbergia sissoo has a unique property in droughts of 3-4 months. Soils range from pure that it hardly regenerates under the old mother sand and gravel to rich alluvium of river banks; trees. It is pioneer in nature and often grows in it can grow in slightly saline soils. Seedlings are clumps in new well-drained alluvial sites near shade intolerant (Nadkarni 1954 & Sheikh river/stream beds. The natural D. sissoo 1989), but mature trees can tolerate moderate populations have come under considerable shade and the species has a low salt tolerance pressure from human disturbance. Very few

- 139 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

remnant populations still exist in the riverine 244, ton ha-1, respectively (Sheikh 1988a). A plains of the Terai and Siwaliks. East Nepal had permanent watertable of 7 m below the soil considerable populations until late 1980s, but surface made the site very favorable. Sissoo now they are confined to a few natural patches should therefore be used in areas where a high- in far-western Nepal, in a very threatened state value timber market is available (NAS 1983, (except in Protected Area Systems). Sheikh & Haq 1982 and Sheikh 1988b).

PHYTOMASS PRODUCTION: PROPAGATION AND MANAGEMENT: Conserving of sissoo tree aims at improving 1-Propagation methods: the quality of production and services from the Seeds remain viable for only a few months operational plantations. Nepal's ninth five-year when exposed to air, but can be stored for up to plan emphasizes development of large-scale tree 4 years in sealed containers (Jackson 1987). It is plantations (30000 ha) through participatory not necessary to extract seeds from pods, which planning and implementation. Tree improve- can be broken into one-seeded segments and ment should have an important role to play in sown. Seeds should be soaked in water for 48 such a process, especially when diseases and hours before sowing, and 60-80% germination pests are affecting one of the major plantation can be expected in 1-3 weeks (Jackson 1987). species of the country (ICRAF undated). The seeds are not hard coated and scarification According to the Wealth of India, irrigated is not necessary. Soaking for 24-48 hours in cold water before sowing improves germination. The plantations of sissoo yield fair quantities of seeds (pod segments) are sown in March-April timber and fuel. In irrigated plantations trees in lined raised seed-beds and watered two times may attain a girth of 1.2 m in 25 years. A height every day. Germination starts after about one of 7 m has been reported in 20 months. Based week and is completed after three weeks. When on studies of 40 natural riverine sites, it was the seedlings are about 5 cm tall, they are concluded that 10-year stands yield about 10 m3 transplanted into containers. For production of ha-1, 20-year stands 100 m3 ha-1 (5m3 ha-1 yr-1), stumps, 12-16 months are required in the 30-year stands 210 m3 ha-1 (7 m3ha-1 yr-1), 40- nursery (NFTA 1992). After collection, the pods year stands 280 m3 ha-1 (7 m3 ha-1 yr-1), 50- year are dried in the sun and, broken into segments stands 370 m3 ha-1 (7.5 m3 ha-1 yr-1), and 60-year each containing one seed. The segments are then stands 460 m3 ha-1 (7.5 m3 ha-1 yr-1) (CSIR 1976). cleaned by winnowing to remove empty pieces In Nepal, a 9.5-year stands thinned to 867 trees of pods (Tewari 1994). The seeds are orthodox ha-1 at 6.5 years, produced an annual increment and when properly dried and stored in airtight of 18.1 m3 (Jackson 1987). A 10-year irrigated containers they will retain high viability for plantation in Peshawar, spaced at 2×2, 3×3 and several years even at room temperature, they 4×4 m produced a total wet weight of main live longer if stored at 5°C. stem, branches, leaves and roots of 510, 231 and

- 140 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

While seeds may be sown without seconds respectively. There has been some pretreatment, it is recommended that they be success with tissue culture (Jackson 1987). soaked in water at room temperature for 24-48 hr, inoculated with Rhizobium after soaking and 2-Tree management and silviculture: sown immediately. Sissoo rarely regenerates Sissoo is a fast-growing species of growth under the parent canopy. Natural regeneration rates of 3.7 m in one year, 5.0 m in 3 years, 11.0 is nonetheless abundant along streams and m in 5 years and 15.0 m in 10 years. Plantations riverbanks where the pods have been carried by are established in block or strip plantations at floods. Ripe pods may be collected manually by 1.8×1.8 m to 4×4 m, closer spacing is used for climbing trees and picking them or by shaking straight timber of good quality. When the the branches and picking them from the canopy closes, at about 6 years, 30-40% of the ground. Propagation by root suckers is done stems are thinned to selectively remove best by cutting stems just below the soil surface. suppressed, diseased and badly formed trees. While it is difficult to propagate using stem and Thinning is recommended every 10 years where branch cuttings without hormone treatments, the rotation is 30-60 years. There is evidence exogenous application of auxins (IAA, IBA and that the stumps begin to lose vigour after 2 or 3 NAA) have been found to improve survival and rotations when managed as a coppice crop. It growth rates (Taylor & Macdicken 1990). coppices vigorously up to about 20-year (White Stump planting is widely employed in 1990). In Pakistan at spacing of 4×4 m, 3×3 m irrigated plantations in India. Trenches are dug and 2×2 m, sissoo height and diameter at the 1.5 m apart, earth thrown a little away from the breast height (dbh) after 6 years were 8.4 m and trenches and the berms used for sowing seed or 11.3 cm, 8.7 m and 10.1 cm, and 8.7 m and 8.6 pod segments (Duke & Wain 1981). Stumps are cm, respectively (Sheikh 1984). Differences were planted in spring, not earlier than the third not significant, but the 2×2 m spacing produced week of March or April. Where subsoil water is trees with fewer branches and more fuel wood. low or rainfall poor and uncertain, irrigation is After 9 years, height and dbh for the three essential (Gohl 1981). Stumps are planted along spacing were 15.1 m and 18.9 cm, 13.4 and 15.6 trenches or on berms of pits and the field are cm, and 13.9 m and 14.2 cm. irrigated. Under proper irrigation, sissoo roots Thorough weeding is important during the tap the subsoil water within 2 years (Kirtikar & first 2-3 years. In a trial at Adabhar (Nepal), Basu 1975). Wood cuttings planted in May and mean height at 18 months was 3.8 m in fully June failed completely, while those planted in cultivated plots and 1.3 m when weeding was August achieved up to 20% success (Vidaeovic confined to a 50 cm diameter circle around the 1968). When summer planted, Pain and Roy plants (Jackson 1987). Protection against (1981) reported 100, 80 and 60% rooting success browsing animals and fire also is essential if the with IBA, NAA and IPA treatment for 30 plant is to become a tree. Irrigation is very

- 141 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

important for establishment of sissoo in Sissoo tolerates disease, drought, frost, semiarid areas. Sissoo should be able to tap sub- insects, porous soils, salt, sand, savanna, sewage, soil water within a couple of years if irrigated and wind. On the clay soils, its growth is properly. Shallow and frequent irrigation or stunted. Seed storage behavior is orthodox; constant flooding induces superficial root viability is maintained for 4 years in hermetic formation. Fertilization with various storage and 1-2 years when stored in airtight combinations and amounts of NPK did not show containers under dry cool conditions of 5-22°C significant effects on dbh or height over 5-6 (Vogt 1995). Dalbergia sissoo reproduces years on a rich soil (Sheikh & Cheema 1986). through seed production and vegetatively Phosphate would normally be expected to through suckers arising from the root system promote early growth on poor soils. (Fig. 10).

- 142 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Fig. 10: Dalbergia sissoo reproducing by suckers arising from the root system

- 143 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

The pollination mechanism is theorized to between populations and for the single be through insects. Regeneration is rare under population over time (White 1994). Ripe fruits the shade of the parent canopy and seed can be harvested from December to March. The dispersal is through wind or water (ICRAF fruits should be collected from the tree by undated). In North America, Dalbergia sissoo climbing or by shaking the fruits onto a begins to produce flowers after nine months, tarpaulin on the ground. It is not advisable to with flowering closely associated with leaf flush collect from the ground as the seeds are often in April-May (Painter 2006). The mature pods infected. 1.25 kg pods contain about 1 kg seed are persistent on the tree for 7-8 months; (Jøker 2002). however seed remains viable for a few months once exposed to air, and germinates in 1-3 ESTS AND DISEASES: weeks during spring (Sheikh 1989). Trees reach leaf defoliator Plecoptera reflexa, leaf roller maturity around 19-21 years of age (Sangha & Dichmeria eridantis, wood borer Stromartium Jalota 2005) with a natural rotation of about 60 barbatum, powder post beetles Sinoxylon anale years (Sharma et al., 2000). The small bisexual and Lyctus africanus are insects that have been flowers are borne on small branches from the reported to cause considerable damage. The leaf axis. Flowering follows leaf flushing; leaves fungus, Ganoderma lucidum, which causes root fall and young flower buds appear with new and butt rot, is common. Fusarium solani and leaves followed by complete pod formation and Polyporus gilvus cause similar diseases. Sissoo maturity. Mature pods remain attached to the suffers minor damage from two foliage rusts tree for 7-8 months and are then dispersed by and a powdery mildew (Jackson 1987). Leaf wind and water (Kamaluddin 1995). diseases include the powdery mildew fungus, Within the area of natural distribution, the leaf spot Cercospora sissoo, leaf blight fungus leaves are shed in November-December and new Colletotrichum sissoo, and leaf wilt Fusarium leaves appear in January-February. The first solani dalbergiae. Wood pathogens recorded flowers appear together with the new leaves and include wood rot fungus Daedalea flavida and the flowers open in March-April. By the end of stump rot fungus Fomes durissimus (Singh April, young green pods appear, and in October 1982). Brachytrypes portentosus causes nursery when the dry season sets-in the fruits begin to damage and termites attack young trees. ripen (Tewari 1994). Seed production starts Parasitic plants reported to cause considerable when the trees are 3-4 years old and normally a damage to sissoo include Loranthus longiflorus good crop is produced every year with yields of and Tapinenthus dodoneifolius in alluvial 1-3 kg per tree. Sissoo being partial selfing and forests; climbers like Dregea volobilis, partial out-crossing, a type of breeding system Cryptolepis buchanani and Acacia pennata cause often found in pioneer species. The rate of out- the same damage. It is reported to have crossing has been estimated to 60-90%, it varies pesticidal properties. Aqueous extracts from the

- 144 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

leaves, stems and roots inhibit the reproduction, sissoo are Fusarium spp., Ganoderma lucidum growth and development of the insect pest and Phellinus gilvus; all of which attack the root Utethesia pulchella. Mixed with Azadirachta and vascular system (Sharma et al., 2000). indica oil cake, sawdust from D. sissoo reduces Several defoliating moths such as Plecoptra egg laying and increases larval mortality in reflexa and Dichomeris eridantis can cause Melodogyne javanica. Methanol extract from the significant biomass reduction in sissoo. Other roots has insecticidal properties, especially insect species that attack it are Stromartium against Diacrisia obliqua, Spodoptera litura and barbatum, Sinoxylon anale, and Lyctus africanus Argina cubrania. The seeds can be infested by (Sheikh 1989). the pea beetle, Bruchus pisorum. Infestation is initiated in the field, but breeding can continue GOODS AND SERVICES: during storage (White 1994). 1-Folk medicine and medicinal uses: Dalbergia sissoo is reported to be a CONTROL MEASURES: stimulant used in folk medicine and remedies Preventing introduction through strict (Nadkarni 1954). It is a folk remedy for quarantine and inspection stations is the excoriations, gonorrhea, and skin ailments primary preventative measure. Education of the (Duke & Wain 1981). Ayurvedics prescribe the public on identity, impact, and control of the leaf juice for eye ailments, considering the wood species is necessary to ensure public support for and bark abortifacient, anthelmintic, keeping the species from being introduced. antipyretic, apertif, aphrodisiac, expectorant, Research and testing, on what kind of impact and refrigerant. They use the wood and bark and what invasion potential the species has on for anal disorders, blood diseases, burning the environment, will determine if the species sensations, dysentery, dyspepsia, leucoderma, can be safely cultivated in the country and skin ailments. Its wood is used for blood (Langeland & Stocker 2001). In Florida, disorders, burning sensations, eye and nose herbicide applications to the base of the trunk of disorders, scabies, scalding urine, stomach D. sissoo are recommended for control problems, and syphilis. The alterative wood is (Langeland & Stocker 2001). Other chemical used in India for boils, eruptions, leprosy and applications can be made on the cut stump, nausea (Kirtikar & Basu 1975). Its leaves are basal bark or as a stem injection (PIER 2006). boiled and given to animals for bilious disorders There is no available literature on a host specific (Baquar 1989). The tree has many reputed organism that is being researched or tested as a medicinal properties and has been used biological control agent for D. sissoo, however culturally for a variety of ailments including: several species of fungi, insect and bacteria skin diseases, blood diseases, syphilis, stomach cause mortality or reduced growth of the tree. problems, dysentry, nausea, eye and nose Species of fungi that attack and commonly kill

- 145 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

disorders, aphrodisiac, expectorant, among ions and has the potential of removing these others (Duke 1983). The alcohol extract of the heavy metals from industrial and commercial green branches of aerial parts showed a dose- waste water sources (Habib-ur-Rehman et al., dependent inhibitory effect on the motility of 2006). The wood has a high caloric content and rabbit duodenum, pronounced bronchodilation, is an important fuel wood and charcoal source as well as significant anti-inflammatory, (Sheikh 1989). The wood fibers are processed antipyretic, analgesic, and estrogen-like into a pulp that is further made into paper activities (Sarg et al., 1999). An aqueous extract (ICRAF undated). Heartwood yields 5.4% of of the leaves of Dalbergia sissoo has been used oil, which approaches the texture of vaseline for the treatment of gonorrhea in Arabic after cooling. It is suitable as a lubricant for countries (El-Dagwy 1996). Oil obtained from heavy machinery (Browne 1968, ICRAF the seeds is used to cure skin diseases. The undated). The wood is excellent for heating and powdered wood, applied externally as a paste, is cooking. The calorific value of the sapwood is used to treat leprosy and skin diseases (Mbuya about 4900 kcal kg-1, and that of heartwood is et al., 1994). The isoflavones irisolidone, about 5200 kcal kg-1. The sapwood is also used biochanin-A, muningin, tectorigenin, prunetin, for pulp making (White 1994). Its wood is well and prunetin-4-O-galactoside, the flavone nor- and does not warp or split; it is extremely artocarpotin, and ß-amyrin, ß-sitosterol and durable and is one of the timbers least stigmasterol were isolated and identified from susceptible to dry-wood termites in India. Wood the green branches of aerial parts of Dalbergia offers resistance to sawing and cutting but is sissoo using silica gel column chromatography excellent for turnery, takes a good polish and and spectral analysis. Also, 13 fatty acids were finishes to a smooth surface. Its root wood is identified and isoflavonoids and neoflavonoids used for tobacco pipes. In village industry, D. have been reported from D. sissoo (Seshadri sissoo is popular for doors and windows. The 1972, Sharma et al., 1979, 1980 and Ingham et heartwood is extremely durable (the specific al., 1983). gravity is 0.7–0.8 g cm3) and is very resistant to dry-wood termites; but the sapwood is readily 2-Wood uses: attacked by fungi and borers. It is a useful The wood of Dalbergia sissoo is highly source of honey, but the flowers are only lightly durable with excellent finishing color and attached to the flower branch and fall easily. smoothness; used for veneer, furniture, The bees are therefore not able to take full cabinets, panelling, carving, small timber, advantage of the large number of flowers. The plywood, ornamental turnery, tool handles, honey produced is dark amber with a strong sprouting goods and musical instruments flavour. Sulphate pulp from wood is used in (ICRAF undated and Lowry & Seebeck 1997). producing writing and printing paper (Parkash The sawdust works in the absorption of nickel & Hocking 1986).

- 146 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

3-Leaves and young shoot uses: Gohl 1981). The fodder value is highest in April and May, when other sources of green fodder The leaves, young shoots and green pods are are scarce (Tewari 1994). Although this fodder used as good fodder for livestock and grazing has no known undesirable compounds, feeding animals, typically in winter seasons when other green leaves sometimes causes digestive fodder is not available (Sheikh 1989). April to disorders which can be prevented by making May is the best time for the production of high- silage (Jackson 1987). Some ethnic groups in quality fodder (Jackson 1987). Based on dry Cameroon are said to relish eating fresh young weight, the leaves of Dalbegia sissoo in India leaves of sissoo (Nadkarni 1954). contain up to 24.1% crude protein, 4.9% fat, 26.1% crude fiber and 12.0% ash (Table 1, after

Table 1: Nutritive values of Dalbegia sissoo leaves in India (after Gohl 1981) Range and mean values (based on dry weight) Nutritive value Minimum Maximum Mean Crude Protein (%) 12.6 24.1 18.4 Fat (%) 2.0 4.9 3.5 Nitrogen -free extract (%) 42.1 54.8 48.5 Crude Fiber (%) 12.5 26.1 19.3 Ash (%) 6.6 12.0 9.3 Calcium (mg) 840.0 2870.0 1855.0 Phosphorus (100 g-1) 120.0 420.0 270.0

4-Ecological importance: along roadsides and in gardens as an ornamental plant (El-Hadidi & Boulos 1988, El- Sissoo provides numerous services to the Sheikh 1989, 1996 and Gilman & Watson 1993). landscape and environment and is commonly It also has been used for landscaping along the employed in agro-forestry (Lowry & Seebeck sea shores of Galilee (Kayastha 1985). 1997). It is used as a windbreak and shelter belt The leaf litter that accumulates and and as a shade tree in intercropping of decomposes also contributes to soil fertility by orchards, mango, tea and coffee plantations. adding additional nitrogen, potassium, iron, Since it has an aggressive root system and is manganese and organic carbon (Keay 1989). In prone to suckering, it is commonly used for India, plant species diversity was much higher erosion control and soil stabilization along in the native D. sissoo monocultures than in the stream and river banks. It is widely planted in exotic Eucalyptus tereticornis ones (Sangha & its native countries for reforestation programs Jalota 2005). In its native range, sissoo is a host (ICRAF undated and Sharma et al., 2000). It is to a variety of orchid species (ICRAF undated). also valued for its ability to increase soil fertility It is therefore found in a variety of wastelands, through nitrogen fixation and is intercropped like in south Asia, where it is known as a for these reasons as well. Due to its fragrant colonizing species. The sub-Himalayas, the flowers and shade, it is planted in urban areas

- 147 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

homeland of D. sissoo, abound with a variety of Appanah, S., Allard, G. and Amatya, S. M. (eds) orchids, many of which are known throughout (2000): Die-back of Sissoo. Proceedings of the world for their beauty (Parrotta 1989). International Seminar, Kathmandu, Although sissoo trees can negatively effect crop Nepal. Forestry Research Support Progr- production due to competition for nutrients, amme for Asia and the Pacific (FROSH). moisture and light; studies have shown that the FAO, Bangkok. net value of intercropping sissoo and wheat is Bangarwa, K. S. (1996): Sissoo breeding. Agric- higher than wheat monocropping (White 1994). ulture and Forestry Information Centre, The ease of propagation by self-seeding, Hisar, India, 156 pp. coppice, root suckers and stumps and the many Banik, H. (1992): Techniques for raising environmental and socio-economic benefits seedlings of timber, fruit and flowers in makes it one of the most valued tree species by the Nursery. Chandpur Press and farmers in the region (Tewari 1994). Publication. Dhaka, 367 pp. Sissoo is one of the most trees used in Baquar, S. R. (1989): Medical and poisonous greenbelt, which is defined as the mass plants of Pakistan. Printas Karachi, plantation of pollution tolerant trees and shrubs Pakistan. 506 pp. in an area for the purpose of minimizing air Bekele-Tesemma, A., Birnie, A. and Tengnas, B. pollution by filtering, intercepting and (1993): Useful trees and shrubs for absorbing pollutants in an effective manner for Ethiopia: identification, propagation, and improvement of the environment. The management for agricultural and pastoral effectiveness of the greenbelt depends on the communities (Technical handbook). several factors such as climatic conditions, Regional Soil Conservation Unit (RSCU), design, selection of plant species and its Swedish International Development characters and type of pollutants. The Authority (SIDA), 474 pp. importance of greenbelt can be ascertained Beniwal, B. S. and Singh, N. B. (1989): Observa- from the estimate of cleaning capacity of 3.7 ton tion on flowering, fruiting and germina- of CO2 from atmosphere and supply of 2.5 ton tion behaviour of some useful forest of oxygen from one hectare of sissoo woodland plants of Arunachal Pradesh. Ind. (Sharma & Roy 1999). Forester 115: 216-227. Browne, F. G. (1968): Pests and diseases of REFERENCES: forest plantations trees. Clarendon Press, Oxford, 1330 pp. Alam, M. K., Mohiuddin, M. and Guha, M. K. CSIR (Council of Scientific and Industrial (2001): Trees for low lying areas of Research) (1976): The Wealth of India. A Bangladesh. Bangladesh Forest Research dictionary of Indian raw materials & Institute, Chittagong, 98 pp. industrial products. National Institute of

- 148 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Science Communication, Pusa, New Hocking, D. (1993): Trees for Dry lands. Oxford Delhi, India 110-112, 10: 581- 585. & IBH Publishing Co. New Delhi. Duke, J. A. and Wain, K. K. (1981): Medicinal Ingham, J. L., Kokinen, A. and Lounasmaa, M. plants of the world. FAO, Rome, Italy 3: (Eds.) (1983): Progress in the Chemistry 277–295. of Natural Organic Products. Springer, El-Dagwy, A. A. (1996): Encyclopedia for Wien/New York, 43: 1–200. production of aromatic and medicinal Jackson, J. Y. (1987): Manual of Afforestation plants, Vol. 1, Imprimerie Atlas, Cairo. in Nepal. Nepal United Kingdom Forest 192 pp. Research Project, Department of Forest, El-Hadidi, M. N. and Boulos, L. (1988): The Kathmandu, Nepal, 199-214. street tress of Egypt. The American James, A. (2002): Dalbergia sissoo Roxb. ex DC. University in Cairo Press, Cairo, 130 pp. Purdue University, Center for New Crops El–Sheikh, M. A. (1989): A study of the and Plant Products, 311 pp. vegetation environmental relationships of Javaid, A., Bajwa, R. and Anjum, T. (2004): the canal banks of the Middle Delta Identification of some more phenotypes region. M. Sc. Thesis, Tanta University, of Shisham (Dalbergia sissoo Roxb.) and Tanta. 139 pp. their response to dieback and wilt El–Sheikh, M. A. (1996): Ruderal plant diseases. University of the Punjab,

communities of the Nile Delta region. Ph. Quaid-e-Azam Campus, Lahore, Pakistan

D. Thesis, Tanta University, Tanta 189 2 (2): 55-59. pp. Jøker, D. (2002): Dalbergia sissoo Roxb. ex DC. Fahmy, H. (2007): Current status of Dalbergia Seed leaflet, Danida Forest Seed Centre, sissoo Roxb. ex DC. Tree in Nile Delta. Denmark 65: 2 pp.

Environmental Science, Mansoura Kamaluddin M. A. (1995): Genetic improvement University, Egypt. Vol. 33: 23 pp. and propagation strategy for Dalbergia Gohl, B. (1981): Tropical feeds. Feed sissoo in Bangladesh. Bangladesh Journal information summaries and nutritive of Forest Science 24 (2): 54-61. values. FAO Animal Production and Kanak, S. and Sahai, K. (1994): Studies on seed Health Series 12. FAO, Rome (Italy), 665 position and their effect on germination pp. and seedling survival in Dalbergia sissoo Habib-ur-Rehman, Shakirullah, M., Ahmad, I., Roxb. Indian Forester 120: 464-465. Shah, S. and Hameedullah (2006): Kayastha, B. P. (1985): Silvics of the trees of Sorption studies of nickel ions onto Nepal. Community Forest Development sawdust of Dalbergia sissoo. Journal of Project, Kathmandu. the Chinese Chemical Society 53: 1045- Keay, R. W. (1989): Trees of Nigeria. Claredon 1052. Press, Oxford, United Kingdom, 476 pp.

- 149 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Kirtikar, K. R. and Basu, B. D. (1975): Indian NAS (National Academy of Science) (1979): medicinal plants. 4 vols, 2nd ed. Jayyed Tropical legumes: resources for the Press, New Delhi. future. National Academy of Science Langeland, K. A. and Stocker, R. K. (2001): Press, Washington DC, 331 pp. Control of non-native plants in natural NAS (National Academy of Science) (1983): areas of Florida, Dept. of Agronomy, Firewood crops: shrub and tree species Florida Cooperative Extension Service, for energy production. National Academy Institute of Food and Agricultural of Science Press, Washington DC, Vol. II: Sciences, University of Florida, IFAS 92 pp. Publication SP 242, 34 pp. Neelu, L., Lobhiyal, S. and Pangtey, Y. (2002): Lanzara, P. and Pizetti, M. (Edt) (1978): Simon Structure and function of shisham forests and Schuster’s guide to trees. Published in central Himalaya, India: dry matter, by Simon and Schuster, Inc., New York. dynamics. Annals of Botany 89: 41-54. Leloup, M. (1956): Tree planting practices in NFTA (Nitrogen Fixing Tree Association) tropical Africa. FAO Publication, 181-183 (1992): Nitrogen Fixing Tree Research pp. Reports. Hawaii, USA Vol. 10: 32 pp. MacDicken, G. K. (1994): Selection and manag- Othman, E. (1939): Woody trees. Al-Etemad ement of nitrogen fixing trees. Winrock Publishing, Cairo, 199 pp (In Arabic).

International and Bangkok: FAO. Pain, S. K. and Roy, B. K. (1981): A comparative Mbuya, L. P., Msanga, H. P., Ruffo, C. K., study of the root forming effect of IPA, Birnie, A. and Tengnäs, B. (1994): Useful IBA, and NAA on stem cuttings of trees and shrubs for Tanzania: identifica- Dalbergia sissoo. Ind. For. 107(3): 151- tion, propagation and management for 158. agricultural and pastoral communities. Pallewatta, N., Reaser, J. K. and Gutierrez, A. Regional Soil Conservation Unit (RSCU), T. (2003): Invasive alien species in south- Swedish International Development southeast Asia. National Reports and Authority (SIDA). Technical Handbook Directory of Resources, Global Invasive No. 6:510-511. Species Programme, Cape Town, South Nadkarni, K. M. (1954): Indian materia medica. Africa, 111 pp. 3rd edition. Bombay: Popular Book Parkash, R. and Hocking, D. (1986): Some Depot Vol. 1: 432 pp. favorite trees for fuel and fodder. Society Napier, I. and Robbins, M. (1989): Forest Seed for Promotion of Wastelands Develop- and Nursery Practice in Nepal. Nepal-UK ment, New Delhi, India 4: 42-49. Forestry Research Project, Department Parrotta, J. A. (1989): Dalbergia sissoo Roxb. of Forestry and Plant Research, Silvics of forest trees of American tropics. Babarmahal, Kathmandu, Nepal. SO-ITF-SM-24, New Orleans, LA: U.S.

- 150 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Dept. of Agriculture, Forest services, Sheikh, M. I. (1984): Effect of spacing on Forest Experimental Station, 5 pp. growth of Dalbergia sissoo. Tech. Notes 1- Sangha, K. K., and Jalota, R. K. (2005): Value 55 pp; Tech. Note 41: 82-83. of ecological services of exotic Eucalyptus Sheikh, M. I. (1988a): Biomass production from tereticornis and native Dalbergia sissoo shisham plantation at different spacings. tree plantations of North-Western India. For. Res. Serv. Tech. Note 62. Conservation and Society 3 (1): 92-109. Sheikh, M. I. (1988b): Comparative growth of Sarg, T., Ateya, A., Abdel-Ghani, A., Badr, W. four species under irrigated conditions. and Shams, G. (1999): Phytochemical For. Res. Serv. Tech. Note 61. and pharmacological studies of Dalbergia Sheikh, M. I. and Cheema, A. (1986): Effect of sissoo growing in Egypt. Journal of fertilizer on the rate of growth of forest Pharmaceutical Biology 37 (1): 54-62. trees. Final Tech. Rep. For. Inst. PSR: Seshadri, T.R. (1972): Polyphenols of 119 pp. Pterocarpus and Dalbergia woods. Sheikh, M. I. and Haq. R. (1982): Performance Phytochemistry 11: 881–898. of popular and other species in Shaltout, K. H. (2003): The biology of Egyptian conjunction with agricultural crops. Pak. woody perennials. 1. Nitraria retusa Jour. For. 32(2): 72-78. (Forssk.) Asch. Ass. Univ. Bull. Environ. Singh, R. V. (1982): Fodder trees of India. Res. 6(1): 55–71. Oxford & IBH Co. New Delhi, India, 663 Shaltout, K. H., Al-Sodany, Y. M. and Eid, E. pp.

M. (2006): The biology of Egyptian woody Slima, D. F. (2006): Sociological behaviour and perennials. 2. Ipomoea carnea Jacq. Ass. variability among Pluchea dioscoridis Univ. Bull. Environ. Res. 9(1): 75–91. (L.) DC. populations in Nile Delta. M.Sc. Shaltout, K. H., and Slima, D. F. (2007): The Thesis, Menoufia University, Shebin El– biology of Egyptian woody perennials. 3. Kom. 169 pp. Pluchea dioscoridis (L.) DC. Ass. Univ. Taylor, D. A. and MacDicken, K.G. (ed.) (1990): Bull. Environ. Res. 10(1): 85–103. Research on multipurpose tree species in Sharma, A., Chibber, S.C. and Chawla, H. M. Asia. Proceedings of an International (1979): Caviunin 7-O-gentiobioside from Workshop held in Los Baños, Dalbergia sissoo pods. Phytochemistry 18: Philippines. (F/FRED C/o Faculty of 1253. Forestry, Kasetsart University, Kasetsart, Sharma, A., Chibber, S. C. and Chawla, H. M. Bangkok, Thailand, 259 pp. (1980): Isocaviunin 7-gentiobioside, a Tewari, D.N.(1994): A Monograph on Dalbergia new isoflavone glycosides from Dalbergia sissoo Roxb. International Book sissoo. Phytochemistry 19: 715. Distributors, Dehra Dun, India, 202 pp.

- 151 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Thothathn, K. (1987): Taxonomic revision of the Central Florida, Circular EES-41, Florida tribe Dalbergieae of Indian Sub- Cooperative Extension Service, Institute of continent. BSI, Howrah, 37 pp. Food and Agricultural Sciences, University of Florida. http://www.p2pays.org. Troup, R. S. (1921): The silviculture of Indian Duke, J. A. (1983): Dalbergia sissoo Roxb. ex. Tree. Clarendon Press, Oxford 1: 336 pp. DC. Handbook of Energy Crops. Vidacovic, M. (1968): Propagation of shisham Unpublished, Purdue University, Center for Dalbergia sissoo by cuttings. UNDP-FAO. New Crops & Plant Products. www.hort. Pak. For. Res. and Training Program. purdue.edu/newcrop/duke_energy.html. Rep. No. 3. GBIF (Global Biodiversity Information Facility) Vogt, K. (1995): A field guide to the (2007):Species: Dalbergia sissoo. http://www. identification, propagation and uses of europe.gbif.net. common trees and shrubs of dry land, Gilman, E.F. and Watson, D.G. (1993): Dalbergia sissoo Indian Rosewood, Fact sheet Sudan. SOS Sahel International (UK), ST-227, Environmental Horticulture 167 pp. Department, Florida Cooperative Extension White, K. J. (1990): Horticultural varieties and Service, Institute of Food and Agricultural amenity uses of Dalbergia sissoo Roxb. in Sciences, Univeristy of Florida. Nepal. Banko Janakari 2(4): 382-384. http://hort.ufl.edu/ trees/DALSISA. White, K. J. (1994): Domestication and breeding ICRAF (International Center for Research in programme for Dalbergia sissoo Roxb. Agroforestry) undated: Dalbergia sissoo. UNDP/FAO Regional Project Improved www.worldagroforestrycentre.org. Productivity of Manmade Forest through Lowry, J.B. and Seebeck, J. (1997): The potential for tropical agroforestry in wood Technological Advances in Tree Breeding and animal feed production. Rural industries and Propagation. RAS/91/004, Ecosystem research and development corporation, Research and Development Bureau Publication No. 97/73. http://www.rirdc. (ERDB), Philippines. gov.au. Zabala, N.Q. (1990): Plantation Silviculture. Painter, D. (2006): Dalbergia sissoo, Master FAO of the UNO part of projects Gardeners of the University of Arizona. Pima BGD/85/011. Field Document No. 19. County Cooperative Extension, Arizona Rome, Italy, 236 pp. Board of Regents. http://cals.arizona.edu /pima/gardening/aridplants/Dalbergia_sissoo. ONLINE REFERENCES: html. PIER (Pacific Island Ecosystems at Risk) AWC (Australian Weed Committee), undated. (2006): Dalbergia sissoo, http://www.hear.org/ Weed Identification, Himalayan Rain tree, pier/species/dalbergia_sissoo.html. Dalbergia sissoo. http://www.weeds.org. Sharma, M. K., Singal, R. M. and Pokhriyal, T. Black, R.J. and Meerow, A.W. (1993): C. (2000): Dalbergia sissoo in India. Enviroscaping to Conserve Energy: Trees for

- 152 - Ass. Univ. Bull. Environ. Res. Vol. 14 No. 2, October 2011

Proceedings of the Sub-Regional Seminar 72110-9370, USA. http://food-security.info/ "Die-Back of Sissoo (Dalbergia sissoo). Food food/d_ sissoo.html. and Agriculture Organization of the United USDA-NRCS (Natural Resource Conservation Nations (FAO), FAO Corporate Document Service) (2007): Dalbergia sissoo. The plants Repository. http://www.fao. org. database Version 3.5, National Plant Data Sharma, S.C. and Roy, R.K. (1999): Center, Baton Rouge. http://plants.usda.gov. Bioremediation of Urban Environmental Wu, S. H., Chaw, S. M. and Rejmánek, M. Pollution by ornamentals. http://isebindia. (2003): Naturalized Fabaceae (Leguminosae) com/95_99/99-10-5.html. species in Taiwan: the first approximation. Sheikh, M. I. (1989): NFT Highlights Sissoo- Botanical Bulletin of Academia Sinica 44 59- The Versatile Rosewood, NFTA 89-07, forest, 66. http://ejournal.sinica.edu.

farm, and community tree network (FACT Net), Winrock International, Morrilton, AR,

بيولوجية النباتات الخشبية المعمرة في مصر 4- نبات السرسوع .Dalbergia sissoo Roxb. ex DC كمال شلتوت، عمرو قشطه

قسم النبات – كلية العلوم - جامعة طنطا - مصر

يهدد ه ادد ا ال م قال المرجعددإ لددإ اسددالط ا المراجددم المالعلبددة ببيولوجيددة نبددات السرسددواا ويعالبددر الرابددم دد سلسلة من البحوث المرجعية الال الالناول بيولوجيدة النباالدات البيدبية المعمدر د مصدر. مثدل اد ا ال راسدات الوجد ااالمام علمدا بيولوجيدة النبدات المصدريينا لدي بدط نحدو اسدالكمال الدنبي د المعلومدات عدن جماعدات النباالدات البرية المصريةا ولكن أيضاً لالح يث الب يم منها. مجال الالوزيم الطبيع له ا النبدات ادو سدجب جبدال الهيماميدا مدن يرق أ غانسالان عبر باكسالان والهند لدإ نيبدال. قدام بدراايم بايدا د عهد محمد علد بإ بدال اد ا النبدات لمصدر ألغراض الزينة واألبيابا وق أصبب من النباالات المالجنسة مصدر حيدث يغدزو حدواه البندواتا جواندب الطدرقا مجالرق الطرقا حواه الحبول والمصاره. ييالمل ا ا المبال علإ معلومات عن الصنيه والسمية نبات السرسواا الوزيع العالم والمحل ا يكل واليدريب النبدداتا بي ددة النبدداتا الكاللددة الحيددةا الالكدداثر وا ار ا امسددالب امات امقالصددا ية والطبيددة والبي يددة. وقدد ل اسددالط ا المراجددم علددإ وجددو نبددي دد المعلومددات المالعلبددة بددالالغير دد سدديولوجية النبددات والركيبدد الددوراث علدد امالدد ا الال رجات البي ية المصاحبة للنباتا كما أن البييم الالغيرات علإ مسالوى األيكال البي ية لجماعدات اد ا النبدات الحالدا أيضاً ل ارسات مسالببلية.

- 153 -