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Vol.4 (6), pp. 116-121, June 2016 ISSN 2354-4147 Article Number: DRJA15232192 Copyright © 2016 Author(s) retain the copyright of this article Direct Research Journal of Agriculture and Food Science http://directresearchpublisher.org/aboutjournal/drjafs

Research Paper

Morphometrical and Histological Study of the Female Reproductive Tract of the Japanese (Coturnixcoturnix japonica ) in Jos, Plateau State, Nigeria

Y. B. Majama 1, G. D. Mshelia 2, J. R. Lawal 3*, M. Zakariah 1, A. C. Charles 1; D. A. Bwala 1, Y. A. Gazali 1 and W. A. Kachamai 1

1Department of Veterinary , University of Maiduguri, P.M.B 1069, Maiduguri, Borno State, Nigeria. 2Department of Veterinary Surgery and Theriogenology, University of Maiduguri, P.M.B 1069, Maiduguri, Borno State, Nigeria. 3Department of Veterinary Medicine, University of Maiduguri, P.M.B 1069, Maiduguri, Borno State, Nigeria. *Corresponding author E-mail: [email protected]

Received 12 April 2016; Accepted 11 May, 2016

This study was carried out to determine the gross studies, the magnum showed well-developed mucosal folds morphometric and histological features of the female and a ciliated psuedostratified epithelium with its lamina reproductive tract of the ( Coturnixcoturnix propria abundantly filled with tubular glands. The isthmus japonica ) from ten apparently healthy adult laying . also showed a ciliated epithelium. The mucosa of the uterus The had a mean body weight of 143.70±10.03 g. The and vagina has ciliated pseudostratified and thrown into mean weight and length of the reproductive tract of female leaf-shaped folds but that of the vagina has numerous, tall, Japanese quail from this study were 7.09±2.65 g and narrow folds bearing many small secondary folds. In 21.13±1.80 cm respectively. The mean length of the various conclusion, it can be stated that the morphometrical and segments of the oviduct obtained from this research was histologic observations from this study provides a baseline 11.69±0.35 cm, 4.18±0.30 cm, 3.94±0.20 cm and 1.34±0.14 data for the female reproductive tract of the adult japanese cm for the magnum, isthmus, uterus and vagina quail. respectively. This study obtained the measurement for the external diameters of the various segment of the oviduct of the female Japanese quail to be 0.56±0.21 cm, 0.33±0.02 Key words : Female reproductive tract, gross biometry cm, 1.19±0.08 cm and 0.41±0.03 cm for the magnum, features, Japanese quail ( Coturnixcoturnix japonica ), Jos, isthmus, uterus and vagina respectively. In the histological Plateau State.

INTRODUCTION

Quail is any of 130 species of small, short-tailed game old world quail are classified in either of two subfamilies, birds of the family (Order ), or while 36 species of the new resembling but generally smaller and less world quail consisting the subfamily Odontophorinae, robust (Hubrecht and Kirkwood, 2010). The 95 species of more nearly resemble old world partridges (Farhan, 2013). Direct Res. J. Agric. Food. Sci. 117

The scientific designation of Japanese quail is quails provides a very useful tool in understanding Coturnixcoturnix japonica , different from the common several physiological and reproductive phenomena of quail (Thear, 2003; Mizutani, 2003). The Japanese quail ; this study provides the scientific basis for the proper is similar in appearance to the European . understanding and identification of any abnormality in the Overall, they are dark brown with mottling above and reproductive tract of the Japanese quail. Therefore, this lighter brown underneath. They have a whitish stripe present study aimed to establish a comparative base – above the eye on the side of the head (Hoffmann, 1988; line data on the normal dimensions of different segments Johnsgard, 1988). of the reproductive tract of the female Japanese quail and The morphology of the Japanese quail differs also provide a base – line data on the histology of the depending on its stage in life; both male and female different segments of the reproductive tract of the female chicks exhibit the same kind of and coloring Japanese quail. (Hubrecht and Kirkwood, 2010). The head are tawny in color, with small black patches littering the area above the (Hubrecht and Kirkwood, 2010). The wings and MATERIALS AND METHODS the back of the chick are pale brown; the back may also have four brown stripes running along its length Sample Collection (Hubrecht and Kirkwood, 2010). It has been reported that the plumage of the Japanese A total of ten adult female Japanese quail quail is sexually dimorphic, allowing the sexes to be (Coturnixcoturnix japonica) were obtained from the distinguished from one another (Mills et al ., 1997; National Veterinary Research instutute (NVRI) Vom, Hubrecht and Kirkwood, 2010). Plateau State, Nigeria. The birds were transported to the The Japanese quail which was introduced into Nigeria Department of Veterinary Anatomy Postgraduate in 1992, have since gained tremendous interest among research Laboratory, Faculty of Veterinary Medicine, Nigerians populace especially because of its highly University of Maiduguri, Borno State, Nigeria. The birds prolific nature, short generation interval, fast growth rate were allowed to acclimatize during which water and feed and less susceptibility to common poultry diseases were given ad-libitum. (Haruna et al ., 1997; Edache et al ., 2005; Adeola, 2006). Commercialization of quail bird production is a recent development in Nigeria (Akpan and Nsa, 2009; Akinola Morphommetric measurements and Sese, 2011). The interest in the Japanese quail as a research Proir to being sacrificed by slaughter, body weight of the was greatly increased after 1957 due to groups at the each live selected quails were taken using an electronic university of California and Auburn University who balance. The body weight of the birds were taken using a proposed its value in biomedical research. It is now container with cover, the weight of the container was first widely used for research purposes in federal and state measured and recorded after which the birds were put tertiary institutions, as well as private Laboratories into the container and their weights recorded by (Ainsworth et al ., 2010). Fields in which quail is widely subtracting the weight of the container from the reading utilized include: genetics, nutrition, physiology, pathology, on the electronic balance. The whole female reproductive embryology, oncology, behavior and the toxicities of tract (ovary to cloaca) was gentle but quickly dissected pesticides (Ainsworth et al ., 2010). out and then weight taken. The sample was stretched on It has been established that Japanese quail mature in a paper and the length (magnum to vagina) and the about 6 weeks and are usually in full laying external diameter was taken. The weights were recorded production by 50 days of age with hens laying up to 200 – in grams (g) using the electronic digital balance while 300 in their first year of lay (NRC,1991; Smith, length and the external diameter of the sample was 2001). measured in centimeters(cm) using ruler, thread and Quails breeds population are variable throughout the vernier callipers. world, but there are no reliable population estimate exists, although the species was considered to be fairly common in China (Del Hoyo et al ., 1994), declines Histological study appear to have occurred in Loas (Duckworth, 2009) and Japan (Okuyama, 2004). There are fears that the species Tissue processing has undergone a significant decline in overall population (Del Hoyo et al ., 1994; Duckworth, 2009). However, there The reproductive tract sample was kept in Boiun’s fluid as is no reliable population estimates exist in Nigeria soon as the morphometric measurements were taken. It (Okuyama, 2004). was retained in the Boiun’s fluid for twenty four hours and The morphometric and histological study of the then transferred to 70% alcohol for 48 h. Small fragments anatomical structure of the female reproductive tract of (about 0.5 cm) of the ovaries and the various segments Majama et al. 118

DISCUSSION of the oviduct (magnum, isthmus, uterus and Vagina) to be embedded were bathed by successsfully passing The present study of the biometrics of the female them through graded series of alcohol to ensure proper reproductive tract of the Japanese quail provided in-vitro dehydration and removal of Bouin’s stain. measurements and this was first of its kind. Ovarian The tissue fragments were cleared in xylene for 4 h. measurement were not taken but it was observed that the Due to the fragile nature of the specimen, graded mixture ovary contains follicles which are masses of yellowish, of clearing agent (xylene) and melted paraffin wax was rounded objects depending on stalks. This finding used as an intermediate step before embedding in concided with similar observation of the ovary by Thomas paraffin wax. (2014) and PoultryHub (2014) (Figure 1). The mean The tissue impregnated with the mixture of xylene and weight and length (magnum to vagina) of the melted paraffin wax was placed in the oven at 63°C for 3 reproductive tract of female Japanese quail from this h in sample bottles and then transferred to pure paraffin study is 7.09±2.65 g and 21.13±1.80 cm respectively wax at 63°C for 8 h. (Table 1). The tissue fragments having been completely impregnated with paraffin wax were mounted on wooden chocks while orienting the specimen in the mould to ensure it is being cut in the right plane. Cooling of the Table 1. Biometric data of the female reproductive tract of mass at 4°C in a refrigerator to improve solidification. The the Japanese quail showing the Mean and Standard tissues were sectioned using a rotary microtome to a deviation (SD). thickness of 5 µm. Parameters Organs Mean± SD The sections were floated in a water bath at 45°C and Weight (g) Live weight 143.70±10.3 transferred to a glass slides to be stained as described by Whole tract 7.09±2.65 Carson (1990) and Luna (1992). Length (cm) Magnum to vagina 21.13±1.80 Magnum 11.69±1.12 Isthmus 4.18±0.96 Uterus 3.94±0.65 Staining protocol Vagina 1.34±0.43

External diameter (cm) Magnum 0.56±0.07 The tissues were stained with Hematoxylin and Eosin (H Isthmus 0.33±0.75 & E) as previously described by Luna (1992). Uterus 1.19±0.25 Vagina 0.41±0.10

Light microscopy From the result of this study, it was also observed that there was a positive correlation between the live body The histological slides were studied using DAI – 180 weight of the female Japanese quail to the weight of the Digital biological microscpe® at x40, x100, x400 whole tract and the length of the oviduct. However, magnifications. And relevant photomicrograph of the Fitzgerald (1969) in a similar study reported that the various sections were taken. length of the oviduct in is longer on a total bird weight basis than other avian species. The mean length of the various segment of the oviduct Statistical analysis obtained from this research is 11.69±0.35 cm, 4.18±0.30 cm, 3.94±0.20 cm and 1.34±0.14 cm for the magnum, The data generated were statistically analyzed using IBM isthmus, uterus and vagina respectively. This finding SPSS ® version 20 statistical package. contrast the result presented by Fitzgerald (1969) and Johnson (1986) that reported slightly different values. They reported slightly higher values for the RESULTS magnum(13±1 cm) and isthmus(6±2 cm) while slightly lower values were reported for the uterus(2±1 cm) and The mean and standard deviation(SD) of the vagina(1 cm). These observed variations may be due morphometics data of the female reproductive tract of the environment, physiological state (active laying adult) and ten adult Japanese quail are shown in Table 1. The result different coturnix species. showed that the birds had a mean body weight of The study also revealed the measurement for the 143.70±10.03 g and weight of the dissected reproductive external diameters of the various segment of the oviduct tract(whole tract) to be 7.09±2.65 g. of the female Japanese quail to be 0.56±0.21 cm, The longest segment and shortest segment of the 0.33±0.02 cm, 1.19±0.08 cm and 0.41±0.03 cm for the oviduct is the magnum and vagina with a mean length of magnum, isthmus, uterus and vagina respectively. 11.69±1.12 cm and 1.34±0.43 cm respectively. The Sufficient reported literature and research work is uterus had a wider diameter of 1.19±0.25 cm and the unavialable for the comparative analysis with other isthmus with a smaller diameter of 0.33±0.75 cm. studies on this aspect of the biometrics of the female Direct Res. J. Agric. Food. Sci. 119

f f f

f m

A B

Figure 1 . Photomicrograph of quail ovary H&E x100. (A) Showing medulla (M) and eggs (f) at various stages of development and (B) showing concentric layers of the ovary.

ciliation

Lumen G TM M

A B

Figure 2. Photomicrograph of quail magnum H & E. (A)x 40 showing the lumen and arrow showing deep crypt and (B) x400 showing strongly basophilic glands (G), Tunica muscularis (TM) and arrow showing ciliation of the epithelium.

quail. This research happened to be first of it kind to concentric layers of the ovarian follicles can be outline the baseline information external diameters of the distinguished as seen in (Figure 1A and B) of the various segment of the oviduct of the female Japanese photomicrograph of the ovary. Similar observations were quail. As with other species of birds, fertilization of the also reported by Anonymous, (2010). In addition the egg takes place in the oviduct, usually by sperm that magnum showed well-developed mucosal folds and a have been stored in the lower end of the tract and ciliated psuedstratified epithelium with its lamina propria squeezed out by the passage of an egg. They then swim abundantly filled with tubular glands. More over, the up the tract and meet the oncoming next egg. isthmus also showed a ciliated epithelium. The mucosa of Contractions of the muscular oviduct and body the shell gland and vagina has ciliated pseudostratified movements help shove the egg along (Ogwuegbu and and thrown into leaf-shaped folds but that of the vagina Aire, 1990; Aughey and Frye, 2001). has numerous, tall, narrow folds bearing many small Histological study of the ovary from this study revealed secondary folds. These obsevations can be seen in the that the ovary is made up of a cortex and medulla. A various photomicrograph of the segments of the oviduct membranagranulosa surrounds the oocyte (follicles) that as represented in (Figures 1 to 5). These observations are at various stages of develpoment. Also the different agrees with similar report of various researchers on the Majama et al. 120

CF

G G

TM TM Bv

A B

Figure 3. Photomicrograph of quail isthmus H & E. (A) x100 and (B) x400 showing deep crypt (CF), arrow showing Blood vessel (Bv), strongly basophilic glands (G), Tunicamuscularis (TM).

Ep

G

TM

A B

Figure 4. Photomicrograph of quail Uterus H & E. (A) x100 arrow showing mucosal folds and, (B) x 400 arrow showing Epithelium of the uterus, strongly basophilic glands (G), Tunica muscularis (TM).

ciliation

TM

P S

A B

Figure 5. Photomicrograph of quail Vagina H & E. (A) x 40 showing numerous tall, narrow mucosal folds and Tunica muscularis (TM). (B) x 400 arrow showing ciliation of Epithelium of the vagina, primary mucosal folds (P) and secondary mucosal folds(S). Direct Res. J. Agric. Food. Sci. 121

histology of the quail bird and other avian species. Fitzgerald TC (1969). The coturnix Quail Anatomy and Histology. The Anonymous (2010) reported similar obsevations on Iowa state University press, Ames.Iowa. Haruna ES, Musa U, Lombin IH, Tat DB, Shamaki DD, Okewale DA magnum, shell gland and vagina, Thomas (2010) also Molokwe JU (1997). Introduction of quails production in Nigeria. Nig. noted similar results on the histology of the hen isthmus Vet. J. 18:104 – 107. while Johnson (1986) observed similar results on the Hoffmann E (1988). Coturnix Quail.Taipei : Yi Hisien publishing co. shell gland on the prominent muscular layer lined with Hubrecht R Kirkwood J (2010). The UFAW Handbook on the care and management of laboratory and other research . John Wiley & tubular and unicellular goblets cells. Mohammadpour and sons Pp : 655 – 674. Keshtmandi (2008) reported results in animal in Johnsgard P (1988). The Quails, Patridges, and of the world. agreement with the findings of this study on the histologic Oxford: Oxford University press. nature of the magnum with other avian species such as Johnson AL (1986). Reproduction in the female. In: Avian physiology, fourth Ed. Sturkie PD, editor. Springer-Verlag, New York. and pigeon. Luna LG (1992). Histopathologic methods and color Atlas of special stains and Tissue Artifacts. American HistolabsInc. Gaithersburg. Mills AD, Crawford LL, Domjan M, Faure JM (1997). “The Behaviour of Conclusion the Japanese or Domestic quail Coturnix japonica” Neuroscience and Biobehavioural reviews 21(3):261:281. Mizutani M (2003).The Japanese quail, laboratory animal research In conclusion, it can be stated that the biometric data and station, Nippon Institute for biologic science, Kobuchizawa, histologic observations from this study provides a Yamanashi, Japan, pp. 408-441. baseline data for the female reproductive tract of the Mohammadpour AA, Keshtmandi M (2008). Histomorphometrical study of infundibulum and magnum in Turkey and Pigeon. World J. adult japanese quail. Though slight variations with the zoology 3(2):47 – 50. values of the morphometric were observed, this study NRC (1991). National Research Council Quail: In microlivestock little provides a baseline data for subsequent studies and known small animals with a promising economic future. National comparative analysis with other avian species. Academy press, Washington DC.Pp 147-155. Ogwuegbu SO, Aire TA (1990).Ultrastructural studies of the magnum and isthmus of the active oviduct of the indigenous helmeted guinea (Numidameleagrisgaleata, Pallas). Veterinary Arhiv. 60: 101- AUTHORS` DECLARATION 108. Okuyama M (2004).Current status of the Japanese quail coturnix Japonica as a game bird. Journal of the Yamashina institute for We declare that this study is an original research by our 35: 18 – 202 research team and we agree to publish it in the Journal. PoultryHub.www.poultryhub.org/physiology/body system/reproductive system Retrieved 3 rd April 2014 nd Smith AJ (2001). Poultry 2 Ed Macmillan publishers Ltd London pp 1- 11. REFERENCES Thear K. (2003). Keeping quail (A guide to domestic and commercial

management), third edition, Broad leys publishing company, London Adeola O (2006). Review of research in nutrient utilization. Inter. J. E9 5EN. Poult. Sci.,5:210-218. Thomas C (2014). VM 8054 Veterinary Histology, Exercise 29: Female Ainsworth SJ, Stanley RL, Evan DJR (2010). Developmental stages of reproductive system the Japanese quail” J. Anatomy 216: 3-15 II:Avianshttp://www.vetmed.vt.edu./education/curriculum/vm8054/labs Akinola LAF, Sese BT (2011). Feeding value of different dietary nd /lab29/lab29.htm Retrieved 2 April 2014. nutrients for Japanese quail ( Coturnixcoturnix japonica ). In proceedings of 36 th Annual conference of Nigeria society of Animal production (eds Adeniji AA, Olatunji EA, Gana ES) pp. 384 – 387. Akpan IA, Nsa EE (2009). Growth performances of growing Japanese quail ( Coturnixcoturnix japonica ) fed diets containing different crude protein levels. Pro.14 th Ann. Conf. of Anim. Sci. Asso.OfNig (ASAN). 14 th – 17 th sept LAUTECH, Ogbomoso, Nigeria, pp 553 – 555 Anonymous (2010). Comparative Histology VANB 602; Histology laboratory manual, University of Pennslyvania, 2010 cal.vet.upenn/projects/histologylabmanualpdf Accessed 2 nd April 2014 Aughey E, Frye FL (2001). Comparative veterinary histology with clinical correlates. Manson Publishing. Carson FL (1990). Histopathology A self-instructional Text, 2 nd Ed, ASCP press, Chicago. Del Hoyo J, Eliott A, Sargatal J (1994). Handbook of the birds of the world Vol. 2: New world vultures to . Lynx Edicions, Barcelona, Spain Duckworth JW (2009). Recent observations of Galliformes in degraded parts of Loas.Galliformed 1: 18-20 Edache JA, Musa U, Haruna RS, karsin PD, Esiliono JO, Jibrin II (2005). Calcium requirement of Japanese quails ( Coturnixcoturnix japonica ) chicks in Plateau state, Nigeria; Nig. J. An. prod. 32:236- 252. Farhan S (2013). The poultry Guide. Quick review of raising Quail birds. Thepoultryguide.com/quick-view-of-raising-quail-birds.