evaluation of the microbial quality of the river waters at branch,

Item Type Journal Contribution

Authors Sabae, S.Z.; Rabeh, S.A.

Citation Egyptian journal of aquatic research, 33(1). p. 301-311

Publisher National Institute of Oceanography and Fisheries, Egypt

Download date 25/09/2021 03:45:15

Link to Item http://hdl.handle.net/1834/1899 EGYPTIAN JOURNAL OF AQUATIC RESEARCH ISSN: 1687-4285 VOL. 33 NO. 1, 2007: 301-311

EVALUATION OF THE MICROBIAL QUALITY OF THE RIVER NILE WATERS AT DAMIETTA BRANCH, EGYPT

SHAWKY Z. SABAE AND SALEH A. RABEH

National Institute of Oceanography and Fisheris [email protected]

Key words: Indicator bacteria, pathogenic bacteria, pollution, Damietta Branch, River Nile

ABSTRACT

This study aims to the evaluation of the microbial quality of the River Nile water at Damietta Branch. Water samples were taken from the branch, during the period from autumn, 2005 till the summer of 2006. Some environmental parameters such as temperature, transparency, depth and pH were monitored. In addition, the bacteriological analyses involved total viable bacterial counts (TVBCs), total coliforms (TC) and estimation of faecal coliforms (FC), faecal streptococci and pathogenic bacteria. The results of physicochemical parameters showed that the temperature values varied from 17°C to 25°C and the transparency from 40cm to 220cm. Whereas, the depths varied from 3m to 25m and pH values from 7.24 to 8.44. The bacteriological analyses showed that TVBCs ranged from 10.8x107 to 150 x107 cfu/ml and from 8.8x107 to152 x107cfu/ml at 22°C and 37°C, respectively. However, maximum counts were recorded during summer and the minimal were detected in winter. The results of the faecal indicators counts revealed that their densities increased from up- to down stream. The pathogenic bacteria were identified using API 20E strip system (BioMereux). One hundred pathogenic bacterial isolates representing eleven genera were identified to species level. This included, the following suggested names: Esherichia coli (16%), Klebsiella pneumoniae (14%), Pseudomonas aeruginosa (12%), Pseudomonas flourcsence (4%), Salmonella colerasuis (11%), Shigella sp. (9%), Serratia liquefaciens (8%), Proteus vulgaris (8%), Acinetobacter sp.(7%), Brenneria nigrifluens (5%), Flavimonas oryzihabitans (3%) and Chryseomonas lutecla (3%). The results of the present investigation revealed that, the River Nile water at Damietta Branch was subjected to sewage pollution during the study period.

1. INTRODUCTION water sources is widely used (APHA, 1995). Youn- Joo et al. (2002) monitored coliform One of the most important factors of bacteria and E. coli in Lake Marinas (located water pollution is the microbial on the border of Texas and Oklahoma) to contamination; especially with pathogenic determine the microbial pollution in the lake microorganisms. Enteric pathogens are environment. Currently, coliforms and E. coli typically responsible for waterborne sickness are of great importance among bacterial (Karaboze et al., 2003). Contamination of indicators used in water quality definition and water is a serious environmental problem as it health risk (Giannoulis et al., 2005). adversely affects the human health and the Pathogens are a serious concern for biodiversity in the aquatic ecosystem. managers of water resources, because The use of indicator bacteria such as excessive amounts of faecal bacteria in faecal coliforms (FC) and faecal streptococci sewage and urban run-off have been known (FS) for assessment of faecal pollution and to indicate risk of pathogen-induced illnesses possible water quality deterioration in fresh in humans (Fleisher et al., 1998). Several EVALUATION OF THE MICROBIAL QUALITY OF THE RIVER NILE WATERS AT DAMIETTA BRANCH, EGYPT

species of gram-negative bacteria present in were recorded by El-Mongy, (1978); Haeikal, municipal wastewater are pathogenic. This (1994); Abu-Shady et al. (1996); Rabeh, pathogenisty is usually associated with (2000); El-Fadaly et al. (2001); Sabae, (2004) certain components of the cell walls, in and Sabae et al. (2006). The objective of this particular the lipopolysaccharide, also known study was to determine the seasonal as LPS or endotoxin, layer (Baron, 1996). variations in microbial pollution indicators Thus, identification of these pathogenic throughout Damietta Branch and to identify agents in water resources is beneficial for the pathogenic bacteria. controlling and prevention planning of the infectious diseases. THE STUDY AREA In Egypt, the River Nile is the main source of drinking water. The Nile, The River Nile travels along Egypt for unfortunately, receives heavy loads of about 940 km behind the High Dam. After industrial, agricultural and domestic wastes. passing , the Nile pursues a north Drinking water must meet specific criteria westerly direction for about 23 km and and standards to ensure that water supplied to divides at El-Qanater Barrage into two the public is safe and free-from pathogenic branches, each of which runs separately to microorganisms as well as hazardous the Mediterranean Sea, forming the Delta compounds (WHO report, 1993). region between both branches. The western Some studies have been published on the branch is Rossetta branch (239 km in length) physico-chemical characters of the River and the eastern branch is Demietta Branch, Nile. Shaaban-Dessouki et al., 1993; Abdo, about 242 km long. Damietta Branch receives 2004 (a) studied the physical and chemical a large amount of effluents of mostly characters of the River Nile at Damietta untreated agricultural domestic and partially branch. Elewa and Gohar, (1999) studied the treated industrial wastewater (Zyadah, 1996). distribution of some heavy metals in the Damietta Branch is the study area under branch. In addition, studies dealing with the investigation. microbial quality of the River Nile waters

Medit. Sea

7 6

5

4

3 2

1

Fig. (1): map showing the sampling locations at Damietta Branch.

302 SHAWKY Z. SABAE AND SALEH A. RABEH

2. MATERIAL AND METHODS MPN of faecal streptococci was determined using azide dextrose broth at Water samples were collected seasonally 37°C for 48hr. Positive tube was indicated by (from subsurface layer) from seven stations at dense turbidity and confirmed using ethyl Damietta Branch between autumn, 2005 and violet azide dextrose broth incubated at 37°C summer, 2006. Sampling stations are shown for 24hr. The formation of purple button at in the map. the bottom of the tube confirmed the presence Station 1: located at El-Qanater region; of faecal streptococci (APHA, 1995). Station 2: located in front of City; Station 3: located in front of Zifta City; 1.2. ISOLATION AND PURIFICATION Station 4: located in front of City; OF GRAM-NEGATIVE BACTERIA Station 5: located at El- Serw region; Station 6: located before Faraskour Barrage; Station Isolation of gram-negative bacteria in 7: located in front of Damietta City. Dameitta Branch water samples were Water samples were collected using performed using MacConkey agar sterile glass bottles and transported in an ice- supplemented with 0.001 g/L crystal violet box to the laboratory to be analysed within 8 (Hausler and Koontz, 1970). Three hundred hours. and five isolates were purified, screened and the suspected similar ones were grouped for 1. PHYSICOCHEMICAL CHARACTERS the purpose of selection and identification processes. Some environmental parameters were measured in the field e. g. temperature, 1.3. IDENTIFICATION OF SOME transparency, depths and pH. GRAM-NEGATIVE PATHOGENS

1.1. BACTERIOLOGICAL One hundred isolates out of three hundred EXAMINATIONS and five from the examined water samples were subjected to identification by The total viable bacterial counts (TVBCs) biochemical characteristics using API 20E at 22°C and 37°C were determined using the strip system (BioMereux). Each API 20E spread-plate method (APHA, 1995). strip consists of twenty seven wells The number of total and faecal coliforms containing dehydrated media. The isolate to was determined using the most probable be tested was suspended in sterile saline and number (MPN) method. Statistical tables added to each well. The inoculated strip was were used to interpret the results to give the incubated for 16-24h and the colour reactions MPN of the bacteria. From each dilution 1ml were noted either positive or negative. was added to each of triplicate tubes containing 5ml of MacConkey broth. The STATISTICAL ANALYSIS tubes were then incubated at 37°C for 48hr for total coliforms and at 44°C (in water bath) The correlation coefficient (r) between the for 24 hr for faecal coliforms. bacteria and the environmental factors using The positive tubes were streaked on the the computer programme Ms Microsoft Excel Eosin Methylene Blue (EMB) agar plates (ver., 2003). using sterile loop and incubated at 37°C for 24 hr. Microscopic examination was carried out to ensure gram-negative, non- spore forming rods (APHA, 1995).

303 EVALUATION OF THE MICROBIAL QUALITY OF THE RIVER NILE WATERS AT DAMIETTA BRANCH, EGYPT

3. RESULTS AND DISCUSSION positive (r = 0.82). This agrees with those reported by Sabae, 1999. Table (1) shows that the highest The most probable number (MPN) of temperature value (29.7°C) was recorded faecal indicators total coliforms (TC), faecal during summer, while the lowest one was coliform (FC) and faecal streptococci (FS) (17°C) detected in winter. The results showed are shown in (Table, 3). MPN of TC varied a noticeable seasonal trend of temperature. from 240 to 16x104/100ml and from 40 to However, the variations in water 7500/100ml water for FC. The counts of FS transparency were almost local, without fluctuated between 4 and 2100/100ml water seasonal trends. The highest value of of Damietta Branch. transparency was detected at Talkha (Station, The highest bacterial indicators were 4) and the lowest one was recorded at recorded at Damietta (Station7).This might Damietta (station, 7) this agrees with those be explained by the effect of domestic and reported by Abdo, (2004 b), which might be agricultural wastes discharge from the attributed to the effluents discharged from urbanized surrounding area (Shaaban- Talkha Electric Power Station. Dessouki et al., 1993). The results show that the depths of the As regards seasonal variations showed branch throughout the investigated period that the high counts of bacterial indicators ranged between 3 and 25 m. The pH values (TC, FC and FS) were detected in the warmer of the Nile water were on the alkaline side. seasons (spring and summer), which might be However, the highest pH value (8.44) was attributed to high temperature and the recorded at Damietta (station, 7) and the discharged waste water during this season lowest was detected (7.24) in front of (Abu-Shady et al., 1996 and Isobe et al., Faraskour Barrage (station, 6). The recorded 2004) pH values were favorable for bacterial In accordance with (Chao et al., 2003) multiplication. significant positive correlation (r = 0.74) The results of the total number of bacteria were recorded between the total number of are shown in (Table, 2). The total number of bacteria and various indicator bacteria in the bacteria ranged from 10.8x107 to 150 x107 Nile water. cfu / ml and from 8.8x107 to 152x107 cfu/ ml The ratio FC/FS points to the source of at 22ºC and 37°C, respectively. Their highest faeces whether it is human (>4) or animal number was recorded obtained during (<0.7) (Geldreich, 1974), accordingly, the summer and the lowest one was detected in FC/FS ratio were in the range of 0.19-11, winter. The data showed that there is gradual which means the mixed origin of faecal increase in the bacterial counts from up-to pollution. On the other hand, Niewolak down stream that is from El-Qanater to (1998) reported that the data on absolute Damietta, which might be attributed to values of FC: FS may not reflect the source domestic, sewage and agricultural effluents of contamination in Czarna Hancza River, discharge into Damietta Branch. Poland. The highest bacterial counts were According to the guideline criteria for recorded for both groups (at 22°C and 37°C faecal indicator organisms of (WHO report, during summer. This might be due to the high 1992) which accept the guide values of the temperature prevailing during this season. investigated bacteria up to 500/100 ml for This finding was in accordance with El- total coliforms and 100/100ml for both faecal Fadaly et al. (2001) and Sabae (2004). coliforms and faecal streptococci. The survey Statistically, the correlation between the of the indicator bacteria along the Damietta bacterial counts at 22°C and 37°C were Branch waters revealed that the Nile waters at this branch is subjected to sewage pollution.

304 SHAWKY Z. SABAE AND SALEH A. RABEH

305 EVALUATION OF THE MICROBIAL QUALITY OF THE RIVER NILE WATERS AT DAMIETTA BRANCH, EGYPT

Table (2): The total viable bacterial (TVBCs) x 107 cfu/ml water of the River Nile at Damietta branch.

Seasons Autumn Winter Spring Summer

Stations 22ºC 37ºC 22ºC 37ºC 22ºC 37ºC 22ºC 37ºC

1 25 28 17.5 8.8 30 20 35 31

2 28 27 25 27 34 27 40 38

3 26 28 19.5 21 39 31.6 80 78

4 30 46 10.8 14.6 44 37.2 84 85

5 43 47 20.7 13 43 41 88 90

6 43 50 19.2 13 64 34.8 108 110

7 46 58 18.7 14 76 77 150 152

Table (3): The most probable number (MPN) of total coliforms (TC), faecal coliforms (FC) and faecal streptococci (FS) /100ml water of the River Nile.

306 SHAWKY Z. SABAE AND SALEH A. RABEH

In the present study, the isolates of genus In the present study, isolates of genus 6 1 were differentiated and confirmed by API were identified as Serratia liquefaciens 20E as E. coli (Table, 4), E. coli, the main (Table, 4) and represented 8% of gram- indicator of faecal pollution, constitute 16% negative bacteria isolated from the tested of the identified gram-negative bacteria in the water samples. This bacterium is considered a examined water. This also indicated that the pathogen of fish (McIntosh and Austin, water of Dameitta Branch is subjected to 1990). On the other hand, genus 7 includes 8 sewage pollution. isolates (8%) of gram-negative bacteria The genus Pseudomonas is made up of which were identified as Proteus vulgaris more than 300 species of gram-negative (Table, 4). bacteria. Based on morphological and API Acinetobacter is a member of the family 20E biochemical reactions (Table, 4) most Nisseriaceae that is normally found in soil, members of genus 2 are identified as water and the moist areas of skin. Members Pseudomonas aeruginosa which is an of genus 8 (7 isolates) are oxidase-negative, opportunistic pathogen of humans. In contrast non-motile, non-spore forming and non- to most enterobaceria, this pathogenic fermentative coccobacillus which were bacterium is the most significant example of identified as Acinetobacter sp. (Table, 4). bacteria capable of multiplying in water. Genus (9), Brenneria, includes 5 isolates Thus, P. aeruginosa is common in the tested (5%) which were identified as Brenneria water of Dameitta Branch of River Nile nigrifluens (Table, 4). This genus, saprophyte (12%). On the other hand, P. flourcsence or pathogen, is associated with plants and represented 4% of the identified gram previously isolated from surface water negative bacteria from the examined water. (Geller, 1986). At present, the genus Klebsiella is Three isolates of genus 10 was identified subdivided into 5 species (Podschun, et al., as Flavimonas oryzihabitans, which is gram- 2001). The isolates of genus 3 were identified negative, yellow pigmented, oxidase as Klebsiella pneumoniae according to negative, and non-fermenting bacillus (Table, morphological and biochemical 4). This organism has rarely been implicated characteristics (Table, 4). K. pneumoniae as human and fish pathogen (Anzai et al., represented 14% of the identified gram- 1997). On the other hand, the three isolates of negative bacteria isolated from Dameitta. genus 11 were motile aerobic gram-negative This pathogenic bacterium has been rod with yellow organic pigment, oxidase- previously isolated from surface water negative, non-ferments and identified as (McIntosh and Austin, 1990; Podschun et al., Chryseomonas luteola (Table, 4). This 2001). There are 2200 known species of bacterium has rarely been reported as a Salmonella which cause intestinal infections human bacterial pathogen (Chihab et al., such as sallmonellosis. The species of genus 2004). C. luteola can be distinguished from 4 was classified as Salmonella cholerasuis other yellow pigmented non-fermenters by because of its biochemical characteristics negative-oxidase reaction and from (Table, 4) such pathogenic bacterium enterobacteria by its strict aerobic growth. constituted 11% of gram-negative bacteria The normal habitats of this organism are from the examined water. On the other hand, unclear, but it is frequently found in water isolates of genus 5 were identified as Shigella soil and other damp environments (Freney et sp and represented 9% of the identified gram- al., 1988). negative bacteria. This pathogenic bacterium In conclusion, the water of the River Nile is an invasive pathogen which causes at Damietta Branch is subjected to faecal shigellasis or Shigella-related diarrhea. pollution and monitoring of microbial quality

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of water is a must to control the spreading of pathogens transmitted by contaminated water.

Table (4): API 20E biochemical characteristics pathogenic of Gram-negative bacteria isolated from Damietta Branch of River Nile.

308 SHAWKY Z. SABAE AND SALEH A. RABEH

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