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Characteristics of artificially ejaculated and ductus deferens semen in the guinea-fowl
Animal Reproduction Science, 12 (1986) 69--7.4
69
Elsevier Science Publishers B.V., Amsterdam --Printed in The Netherlands
CHARACTERISTICS OF ARTIFICIALLY EJACULATED AND DUCTUS DEFERENS SEMEN IN THE GUINEA-FOWL
NOBORU FUJIHARA 1, HISAYOSHI NISHIYAMA2 and OSAMU KOGAI
IDepartment of Animal Science, Kyushu University 46-06, Hakozaki, Fukuoka 812 (Japan) ZDepartment of Animal Science, Kyushu Tokai University, Choyo, Aso, Kumamoto 869-14 (Japan) (Accepted 31 October 1985)
ABSTRACT Fujihara, N., Nishiyama, H. and Koga, O., 1986. Characteristics of artificially ejaculated and ductus deferens semen in the guinea-fowl. Anim. Reprod. Sci., 12: 69-74. This study was conducted to determine whether the ductus deferens semen of the guinea-fowl is diluted with a fluid at ejaculation and to locate the region ejecting the fluid. Some of the ejaculated semen, which was collected by the lumbar massage method, gave a slight positive aldose reaction when examined by chromatography in contrast to a negative aldose reaction in the ductus deferens semen. No significant differences were found in the pH value and concentration of spermatozoa between the ejaculated and ductus deferens semen. However, the volume of the ejaculate was significantly (P<0.05) greater than that of the semen contained in the receptacular region which corresponds to the receptacle of the ductus deferens in cocks and drakes. These results suggest that a fluid containing aldose may be added in small amounts to the ductus deferens semen during natural ejaculation. It is probably a transudate from the tissue in the vicinity of the papilla of the ductus deferens.
INTRODUC~ON Guinea-fowl have been i n t r o d u c e d r e c e n t l y to Japan by the p o u l t r y i n d u s t r y . H o w e v e r , i n f o r m a t i o n a b o u t t h e r e p r o d u c t i o n o f t h i s s p e c i e s is sparse. Investigations of the r e p r o d u c t i v e p h y s i o l o g y of the guinea-fowl h a v e so f a r i n c l u d e d w o r k o n t h e c o p u l a t o r y o r g a n ( H a s i m o t o a n d M a t u u r a , 1934; Hasimoto, 1936), paracloacal vascular body and phallus (Sasaki e t al,, 1 9 8 3 ; 1 9 8 4 ) , s p e r m a t o g e n e s i s ( A i r e e t al., 1 9 8 0 ) , s p e r m a t o z o a ( T h u r s t o n e t al., 1 9 8 2 a ) , s e m e n ( L a k e , 1 9 7 8 ; B r i l l a r d a n d R e v i e r s , 1 9 8 1 ; T h u r s t o n e t al., 1 9 8 2 b ) a n d f e r t i l i t y ( O f f i o n g a n d A b e d , 1 9 8 0 ) , b u t l i t t l e w o r k h a s b e e n d o n e o n t h e p r o p e r t i e s o f its s e m e n .
0378-4320]86]$03.50
© 1986 Elsevier Science PUblishers B.V.
70 We have previously reported on possible male accessory reproductive organs in the domestic chicken (Nishiyama, 1955; Nishiyama and Fujishima, 1961; Nishiyama and Ogawa, 1961), duck (Nishiyama et al., 1976; Fujihara et al., 1976; Fujihara and Nishiyama, 1976a,b,c) and turkey t o m (Fujihara and Nishiyama, 1984; Fujihara et al., 1985a,b), which produce fluid added to the ductus deferens semen at the time of natural ejaculation. The purpose of the present experiment was to compare the artificially ejaculated semen with the ductus deferens contents to characterize guineafowl semen, with special reference to the possible addition of the accessory reproductive fluid during natural ejaculation. MATERIALS AND METHODS Mature guinea-fowl were caged individually and reared similarly to chickens under a regular lighting regimen (14L:10D). Semen was collected by the lumbar massage m e t h o d used for the rooster (Burrows and Quinn, 1937). The following properties were examined: pH value, volume of semen, aldose color reaction, concentration of spermatozoa, motility and abnormality of spermatozoa, according to the methods described by Wheeler and Andrews (1943), Lake (1957), Ashizawa et al. (1976), and Nishiyama et al. (1976). A t t e m p t s were made to take consecutive semen collections at intervals of a few minutes to confirm the ejection of fluid other than ductus deferens semen in response to lumbar massage. At the end of the experiment, male guinea-fowl were necropsied and semen from the ductus deferens was collected. Although the receptacle of the ductus deferens (Lake, 1981) in the guinea-fowl was n o t obvious, in contrast to other male birds such as cock, drake and turkey tom, the ductus deferens was divided into three portions; receptacular, posterior and anterior regions and the properties of the contents of these regions were compared with each other and with artificially ejaculated semen. Observations on the cloacal region were made through a duckbill vaginal speculum when the male was anesthetized, and particular attention was paid to the tissue at the vicinity of the papilla of the ductus deferens (TVP). All data were subjected to statistical analysis using analysis of variance followed b y t-test and multiple comparisons (Steel and Torrie, 1980). RESULTS The successful collection of semen by natural mating has been reported in several domestic birds, b u t the authors failed to get guinea-fowl males to copulate. Thus, semen reported here was obtained b y the lumbar massage method. Some of the males produced semen with a negative reaction in aldose chromatography, and some birds ejaculated semen containing slightly detectable aldose. Semen samples obtained from the same bird tended to
71
show a similar degree of aldose color reaction. Degree of aldose color reaction depended u p o n individuals. Ductus deferens semen, in contrast, always showed a negative aldose reaction (Table 1). The pH of the ductus deferens semen tested with brom t h y m o l blue (BTB) was almost the same as that of the ejaculated semen. The volume of the ejaculate was significantly (P<0.05) greater than that of the semen contained in the receptacular region o f the ductus deferens, but the quantity o f semen in the posterior region was much more than that of the ejaculate and of the semen in the anterior ductal region (Table 1). The density of spermatozoa was n o t significantly different between the ejaculate and the ductus deferens semen, particularly that in the receptacular and posterior regions of the ducts. However, there was a slightly higher TABLE 1 C h a r a c t e r i s t i c s o f t h e e j a c u l a t e d a n d d u c t u s d e f e r e n s s e m e n in t h e guinea f o w l Semen source
Birds (No.)
pH
Aldose reaction
Semen v o l u m e (~!)
Spermatozoal concentration (× 109 ceUs/ml)
Ejaculated R e c e p t a c u l a r region Posterior region Anterior region
6 6 6 6
7.0+0.1 al 7.0±0.1 a 7.0+ 0.1 a 7.0+0.1 a
± -
17.8±11.0 b 5.9+ 2,3 a 57.6±19.1 c 13.8±11.8 b
4.14+1.02 a 4.20i0.99 a 4.70+0.94 a 5.12±0.73 a
1Means± S.D. Values w i t h i n a c o l u m n w i t h d i f f e r e n t s u p e r s c r i p t l e t t e r s d i f f e r significantly (P<0.05).
Fig. 1. V e n t r a l cloaca o f t h e m a t u r e g u i n e a - f o w l , o p e n e d a l o n g w i t h t h e dorsal m i d l i n e o f Cloacal wall. G = glandular tissue; LP = lateral phallic b o d y ; P A = papilla; P H = phallic b o d y ; T V P = tissue at t h e vicinity o f t h e papilla o f t h e d u c t u s d e f e r e n s .
72 concentration of spermatozoa in the anterior portion of the ductus deferens (Table 1). The motility of spermatozoa immediately after semen collection was scored 4 to 5 according to the assessment of Wheeler and Andrews (1943), and the proportion of morphologically abnormal spermatozoa ranged from 10 to 30 percent, depending u p o n individuals. Most of the abnormal s p e r m a t o z o a h a d a bent neck. Successive collection of semen was n o t ; possible because the male was n o t able to ejaculate again for a few-minutes. Observations of the cloacal region revealed wminute a m o u n t of transudate from the TVP (Fig. 1 ) ; f i l t e r - p a p e r - p l a c e d on this tissue absorbed fluid which showed a slight positive aldose~reaction. DISCUSSION Male guinea-fowl were n o t successfully trained to copulate, thus the analytical results obtained w i t h s e m e n collected manually in this study m a y n o t reflect t h e characteristics o f na~tural semen. In the turkey, the male produced a frothy fluid only during copulation and none was obtained b y l u m b a r massage (Fujihara and Nishiyama, 1984). On the other hand, the rooster produces transparent fluid in response to lumbar massage as well as during natural mating (Nishiyama, 1955; Nishiy a m a and Fujishima, 1961). A similar reaction w a s d e m o n s t r a t e d in the drake (Nishiyama et al., 1976). Since the volume of semen contained in the receptacular region was significantly less than that of the ejaculate, it is suggested that some semen from the posterior portion of the ductus deferens could be emitted during ejaculation induced artificially. In the rooster, drake and turkey tom, the quantity o f semen in the receptacular region is nearly comparable to the volume of ejaculated semen. These species unlike the guinea-fowl have noticeable dilated receptacle portions of the ducti deferentes. Since no significant differences were found in the concentration of spermatozoa or pH values between the ejaculated and ductus deferens semen, it is unlikely that accessory reproductive fluid is added to the ductus deferens semen at the m o m e n t of ejaculation. However, some of the artificially ejaculated semen samples gave a slight aldose color reaction, suggesting that in these cases a fluid containing aldose was included in the ejaculated semen because the ductus deferens semen did n o t contain this compound. Addition of fluids containing aldose to the ductus deferens semen at the time of ejaculation has so far been demonstrated in the semen collected b y natural copulation in the rooster (Nihiyama and Fujishima, 1961), drake (Nishiyama et al., 1976) and turkey t o m (Fujihara and Nishiyama, 1984). Inspection of the cloacal region through a vaginal speculum revealed that the mucosal region in the vicinity of the papilla was red, presumably because it was rich in engorged blood vessles. This region referred to as TVP in this paper was probably the area producing a fluid containing aldose because it
73 gave a slight positive aldose c o l o r r e a c t i o n in s o m e birds in m a r k e d c o n t r a s t t o t h e negative r e a c t i o n b y fluid f r o m the o t h e r cloacal regions. T h e r e was a d i f f e r e n c e in the degree o f aldose c o l o r r e a c t i o n b e t w e e n t h e e j a c u l a t e d s e m e n and the fluid w h i c h o o z e d o u t f r o m the T V P o n t o a filter p a p e r o r seen w i t h t h e vaginal s p e c u l u m . This m a y be d u e t o the v o l u m e o f fluid t r a n s u d e d at e j a c u l a t i o n d u r i n g high sexual e x c i t e m e n t being greater t h a n was a p p a r e n t b y visual i n s p e c t i o n . T h e T V P in the guinea-fowl m a y be similar in h i s t o l o g y a n d f u n c t i o n t o t h e vascular tissue in t h e r o o s t e r ( L a k e and E1 J a c k , 1 9 6 6 ) , t h e E G R (ejacu l a t o r y g r o o v e region) in t h e d r a k e ( F u j i h a r a e t al., 1 9 7 6 ) and t h e T V P in t u r k e y t o m ( F u j i h a r a e t al., 1 9 8 5 a , b ) . T h e T V P , w h i c h is rich in b l o o d vessels, m a y be a special tissue c o m m o n t o male birds and a s o u r c e o f aldose in t h e c l o a c a d u r i n g ejaculation. T h e e j a c u l a t e d s e m e n o f t h e r o o s t e r (Nishiy a m a , 1 9 5 5 ; N i s h i y a m a a n d Fujishima, 1 9 6 1 ) , d r a k e ( N i s h i y a m a et al., 1 9 7 6 ) and t u r k e y t o m ( F u j i h a r a and N i s h i y a m a , 1 9 8 4 ) c o n t a i n s a b o u t 20% o f a c c e s s o r y r e p r o d u c t i v e fluids in the male t u r k e y and 50 p e r c e n t in the r o o s t e r and drake, d e p e n d i n g u p o n t h e m e t h o d o f collection. This c o u l d r e f l e c t t h e greater cloacal tissue swelling in t h e s e birds at ejaculation. T h u r s t o n e t al. ( 1 9 8 2 b ) m e n t i o n e d t h a t the seminal plasma a m i n o acids and p r o t e i n p r o f i l e s o f t h e guinea-fowl were similar t o t h o s e r e p o r t e d f o r t h e c h i c k e n and t u r k e y . T h e results d e s c r i b e d h e r e suggest t h a t male guinea-fowl a p p e a r t o have, in t h e cloaca, tissue similar t o t h e a c c e s s o r y r e p r o d u c t i v e organs in t h e t u r k e y t o m ( F u j i h a r a e t al., 1 9 8 5 a , b ) t h o u g h f u r t h e r e x p e r i m e n t s are n e e d e d t o s u b s t a n t i a t e the characteristics o f t h e s e m e n c o l l e c t e d b y n a t u r a l c o p u l a t i o n . ACKNOWLEDGEMENTS E x c e l l e n t t e c h n i c a l assistance was p r o v i d e d b y Mr. N. Y o s h i h i r o . REFERENCES Aire, T.A., Olowo-okorun, M.O. and Ayeni, J.S., 1980. The seminiferous epithelium in the guinea fowl (Numida meleagris). Cell Tissue Res., 205: 319--325. Ashizawa, K., Nishiyama, H. and Nagae, T., 1976. Effects of oviducal cells on the survival and fertilizing ability of fowl spermatozoa. J. Reprod. Fertil., 47: 305--311. Brillard, J.P. and Reviers, M., 1981. Testis development and daily sperm output in guineafowl raised under constant daily photoperiods. Reprod. Nutr, Develop., 21 (6B): 1105--1112. Burrows, W.H. and Quinn, J.P., 1937. The collection of spermatozoa from the domestic fowl and turkey. Poultry Sci., 16: 19--24. Fujihara, N. and Nishiyama, H., 1976a. Studies on the accessory reproductive organs in the drake. 3. Ejection mechanism of the fluid from the ejaculatory groove region. Poultry Sci., 55 : 1026--1030. Fujihara, N. and Nishiyama, H., 1976b. Studies on the accessory reproductive organs in the drake. 4. Effects of androgen on the ejaculatory groove region of the drake. Poultry Sci., 55: 1324--1330.
74 Fujihara, N. and Nishiyama, H., 1976c. Studies on the accessory reproductive organs in the drake. 5. Effects of the fluid from the ejaculatory groove region on the spermatozoa of the drake. Poultry Sci., 55: 2415-2420. Fujihara, N. and Nishiyama, H., 1984. Addition to semen of a fluid derived from the cloaca1 region by male turkey. Poultry Sci., 63: 554-557. Fujihara, N., Nishiyama, H. and Nakashima, N., 1976. Studies on the accessory reproductive organs in the drake. 2. Macroscopic and microscopic observations on the cloaca of the drake with special reference to the ejaculatory groove region. Poultry Sci., 55: 927-935. Fujihara, N., Nishiyama, H. and Koga, O., 1985a. Studies on the accessory reproductive organs in the male turkey. Proceedings of 3rd AAAP Animal Science Congress, 6 May-10 May 1985, at Seoul, Korea, Vol. 1: 393-395. Fujihara, N., Nishiyama, H. and Koga, O., 198513. The mechanism of the ejection of frothy fluid from the cloaca in the male turkey. Poultry Sci., 64: 1377-1381. Hasimoto, D., 1936. Development of the phallus in the guinea-fowl (Numida meleagris L.). Jpn. J. Zootech. Sci., 10: 200-204 (in Japanese). Hasimoto, D. and Matuura, H., 1934. Copulatory organ of the guinea-fowl (Numida meleagris L.). Jpn. Zootech. SC!., 7: 209-215 (in Japanese). Lake, P.E., 1957. Fowl semen as collected by the massage method. J. Agric. Sci., 49: 120-127. Lake, P.E., 1978. The principles and practice of semen collection and preservation in birds. Symp. Zool. Sot. Lond. No. 43: 31-49. Lake, P.E., 1981. Male genital organ. In: A.S. King and J. McLelland (Editors), Academic Press, New York, Vol. 2. pp. 1-61. Lake, P.E. and El Jack, M.H., 1966. The origin and composition of fowl semen. In: C. Horton-Smith and E.C. Amoroso (Editors), Physiology of the Domestic Fowl. Oliver and Boyd Ltd., Edinburgh, pp. 44-51. Nishiyama, H., 1955. Studies on the accessory reproductive organs in the cock. J. Fat. Agr. Kyushu Univ., 10: 277-307. Nishiyama, H. and Fujishima, T., 1961. On the ejection of the accessory reproductive fluid of the cock during natural copulation. Memo. Fat. Agr. Kagoshima Univ., 4: 27-42. Nishiyama, H. and Ogawa, K., 1961. On the function of the vascular body, an accessory reproductive organ, of the cock. Jpn. J. Zootech. Sci., 32: 98-98. Nishiyama, H., Nakashima, N. and Fujihara, N., 1976. Studies on the accessory reproductive organs in the drake. 1. Addition to semen of the fluid from the ejaculatory groove region. Poultry Sci., 55: 234-242. Offiong, S.A. and Abed, S.M., 1980. Fertility, hatchability and malformations in guinea fowl embryos as affected by dietary manganese. Br. Poult. Sci., 21: 371-375. Sasaki, H., Nishida, T. and Mochizuki, K., 1983. Intercellular junction of urodeal and phallic epithelial cells in the guinea fowl, Numida meleagris. Jpn. J. Vet. Sci., 45: 313-321. Sasaki, H., Nishida, T., Fujimura, H. and Mochizuki, K., 1984. Vascular system of paracloacal-vascular body in the guinea fowl, Numida meleagris. Jpn. J. Vet, Sci., 46: 425-435. Steel, R.G.D. and Torrie, J.H., 1980. Principles and Procedures of Statistics. A biometrical approach. 2nd. ed. McGraw-Hill, New York, pp. 86-238. Thurston, R.J., Hess, R.A., Hughes, B.L. and Froman, D.P., 1982a. Ultrastructures of the guinea fowl (Numida meleagris) spermatozoa. Poultry Sci., 61: 1738-1743. Thurston, R.J., Hess, R.A., Hughes, B.L. and Froman, D.P., 1982b. Seminal plasma free amino acids and seminal and blood plasma proteins of the guinea fowl (Numida meleagris). Poultry Sci., 61: 1744-1747. Wheeler, N.C. and Andrews, F.N., 1943. Seasonal variation in semen production in the domestic fowl. Poultry Sci., 22: 361-367.
69
Elsevier Science Publishers B.V., Amsterdam --Printed in The Netherlands
CHARACTERISTICS OF ARTIFICIALLY EJACULATED AND DUCTUS DEFERENS SEMEN IN THE GUINEA-FOWL
NOBORU FUJIHARA 1, HISAYOSHI NISHIYAMA2 and OSAMU KOGAI
IDepartment of Animal Science, Kyushu University 46-06, Hakozaki, Fukuoka 812 (Japan) ZDepartment of Animal Science, Kyushu Tokai University, Choyo, Aso, Kumamoto 869-14 (Japan) (Accepted 31 October 1985)
ABSTRACT Fujihara, N., Nishiyama, H. and Koga, O., 1986. Characteristics of artificially ejaculated and ductus deferens semen in the guinea-fowl. Anim. Reprod. Sci., 12: 69-74. This study was conducted to determine whether the ductus deferens semen of the guinea-fowl is diluted with a fluid at ejaculation and to locate the region ejecting the fluid. Some of the ejaculated semen, which was collected by the lumbar massage method, gave a slight positive aldose reaction when examined by chromatography in contrast to a negative aldose reaction in the ductus deferens semen. No significant differences were found in the pH value and concentration of spermatozoa between the ejaculated and ductus deferens semen. However, the volume of the ejaculate was significantly (P<0.05) greater than that of the semen contained in the receptacular region which corresponds to the receptacle of the ductus deferens in cocks and drakes. These results suggest that a fluid containing aldose may be added in small amounts to the ductus deferens semen during natural ejaculation. It is probably a transudate from the tissue in the vicinity of the papilla of the ductus deferens.
INTRODUC~ON Guinea-fowl have been i n t r o d u c e d r e c e n t l y to Japan by the p o u l t r y i n d u s t r y . H o w e v e r , i n f o r m a t i o n a b o u t t h e r e p r o d u c t i o n o f t h i s s p e c i e s is sparse. Investigations of the r e p r o d u c t i v e p h y s i o l o g y of the guinea-fowl h a v e so f a r i n c l u d e d w o r k o n t h e c o p u l a t o r y o r g a n ( H a s i m o t o a n d M a t u u r a , 1934; Hasimoto, 1936), paracloacal vascular body and phallus (Sasaki e t al,, 1 9 8 3 ; 1 9 8 4 ) , s p e r m a t o g e n e s i s ( A i r e e t al., 1 9 8 0 ) , s p e r m a t o z o a ( T h u r s t o n e t al., 1 9 8 2 a ) , s e m e n ( L a k e , 1 9 7 8 ; B r i l l a r d a n d R e v i e r s , 1 9 8 1 ; T h u r s t o n e t al., 1 9 8 2 b ) a n d f e r t i l i t y ( O f f i o n g a n d A b e d , 1 9 8 0 ) , b u t l i t t l e w o r k h a s b e e n d o n e o n t h e p r o p e r t i e s o f its s e m e n .
0378-4320]86]$03.50
© 1986 Elsevier Science PUblishers B.V.
70 We have previously reported on possible male accessory reproductive organs in the domestic chicken (Nishiyama, 1955; Nishiyama and Fujishima, 1961; Nishiyama and Ogawa, 1961), duck (Nishiyama et al., 1976; Fujihara et al., 1976; Fujihara and Nishiyama, 1976a,b,c) and turkey t o m (Fujihara and Nishiyama, 1984; Fujihara et al., 1985a,b), which produce fluid added to the ductus deferens semen at the time of natural ejaculation. The purpose of the present experiment was to compare the artificially ejaculated semen with the ductus deferens contents to characterize guineafowl semen, with special reference to the possible addition of the accessory reproductive fluid during natural ejaculation. MATERIALS AND METHODS Mature guinea-fowl were caged individually and reared similarly to chickens under a regular lighting regimen (14L:10D). Semen was collected by the lumbar massage m e t h o d used for the rooster (Burrows and Quinn, 1937). The following properties were examined: pH value, volume of semen, aldose color reaction, concentration of spermatozoa, motility and abnormality of spermatozoa, according to the methods described by Wheeler and Andrews (1943), Lake (1957), Ashizawa et al. (1976), and Nishiyama et al. (1976). A t t e m p t s were made to take consecutive semen collections at intervals of a few minutes to confirm the ejection of fluid other than ductus deferens semen in response to lumbar massage. At the end of the experiment, male guinea-fowl were necropsied and semen from the ductus deferens was collected. Although the receptacle of the ductus deferens (Lake, 1981) in the guinea-fowl was n o t obvious, in contrast to other male birds such as cock, drake and turkey tom, the ductus deferens was divided into three portions; receptacular, posterior and anterior regions and the properties of the contents of these regions were compared with each other and with artificially ejaculated semen. Observations on the cloacal region were made through a duckbill vaginal speculum when the male was anesthetized, and particular attention was paid to the tissue at the vicinity of the papilla of the ductus deferens (TVP). All data were subjected to statistical analysis using analysis of variance followed b y t-test and multiple comparisons (Steel and Torrie, 1980). RESULTS The successful collection of semen by natural mating has been reported in several domestic birds, b u t the authors failed to get guinea-fowl males to copulate. Thus, semen reported here was obtained b y the lumbar massage method. Some of the males produced semen with a negative reaction in aldose chromatography, and some birds ejaculated semen containing slightly detectable aldose. Semen samples obtained from the same bird tended to
71
show a similar degree of aldose color reaction. Degree of aldose color reaction depended u p o n individuals. Ductus deferens semen, in contrast, always showed a negative aldose reaction (Table 1). The pH of the ductus deferens semen tested with brom t h y m o l blue (BTB) was almost the same as that of the ejaculated semen. The volume of the ejaculate was significantly (P<0.05) greater than that of the semen contained in the receptacular region o f the ductus deferens, but the quantity o f semen in the posterior region was much more than that of the ejaculate and of the semen in the anterior ductal region (Table 1). The density of spermatozoa was n o t significantly different between the ejaculate and the ductus deferens semen, particularly that in the receptacular and posterior regions of the ducts. However, there was a slightly higher TABLE 1 C h a r a c t e r i s t i c s o f t h e e j a c u l a t e d a n d d u c t u s d e f e r e n s s e m e n in t h e guinea f o w l Semen source
Birds (No.)
pH
Aldose reaction
Semen v o l u m e (~!)
Spermatozoal concentration (× 109 ceUs/ml)
Ejaculated R e c e p t a c u l a r region Posterior region Anterior region
6 6 6 6
7.0+0.1 al 7.0±0.1 a 7.0+ 0.1 a 7.0+0.1 a
± -
17.8±11.0 b 5.9+ 2,3 a 57.6±19.1 c 13.8±11.8 b
4.14+1.02 a 4.20i0.99 a 4.70+0.94 a 5.12±0.73 a
1Means± S.D. Values w i t h i n a c o l u m n w i t h d i f f e r e n t s u p e r s c r i p t l e t t e r s d i f f e r significantly (P<0.05).
Fig. 1. V e n t r a l cloaca o f t h e m a t u r e g u i n e a - f o w l , o p e n e d a l o n g w i t h t h e dorsal m i d l i n e o f Cloacal wall. G = glandular tissue; LP = lateral phallic b o d y ; P A = papilla; P H = phallic b o d y ; T V P = tissue at t h e vicinity o f t h e papilla o f t h e d u c t u s d e f e r e n s .
72 concentration of spermatozoa in the anterior portion of the ductus deferens (Table 1). The motility of spermatozoa immediately after semen collection was scored 4 to 5 according to the assessment of Wheeler and Andrews (1943), and the proportion of morphologically abnormal spermatozoa ranged from 10 to 30 percent, depending u p o n individuals. Most of the abnormal s p e r m a t o z o a h a d a bent neck. Successive collection of semen was n o t ; possible because the male was n o t able to ejaculate again for a few-minutes. Observations of the cloacal region revealed wminute a m o u n t of transudate from the TVP (Fig. 1 ) ; f i l t e r - p a p e r - p l a c e d on this tissue absorbed fluid which showed a slight positive aldose~reaction. DISCUSSION Male guinea-fowl were n o t successfully trained to copulate, thus the analytical results obtained w i t h s e m e n collected manually in this study m a y n o t reflect t h e characteristics o f na~tural semen. In the turkey, the male produced a frothy fluid only during copulation and none was obtained b y l u m b a r massage (Fujihara and Nishiyama, 1984). On the other hand, the rooster produces transparent fluid in response to lumbar massage as well as during natural mating (Nishiyama, 1955; Nishiy a m a and Fujishima, 1961). A similar reaction w a s d e m o n s t r a t e d in the drake (Nishiyama et al., 1976). Since the volume of semen contained in the receptacular region was significantly less than that of the ejaculate, it is suggested that some semen from the posterior portion of the ductus deferens could be emitted during ejaculation induced artificially. In the rooster, drake and turkey tom, the quantity o f semen in the receptacular region is nearly comparable to the volume of ejaculated semen. These species unlike the guinea-fowl have noticeable dilated receptacle portions of the ducti deferentes. Since no significant differences were found in the concentration of spermatozoa or pH values between the ejaculated and ductus deferens semen, it is unlikely that accessory reproductive fluid is added to the ductus deferens semen at the m o m e n t of ejaculation. However, some of the artificially ejaculated semen samples gave a slight aldose color reaction, suggesting that in these cases a fluid containing aldose was included in the ejaculated semen because the ductus deferens semen did n o t contain this compound. Addition of fluids containing aldose to the ductus deferens semen at the time of ejaculation has so far been demonstrated in the semen collected b y natural copulation in the rooster (Nihiyama and Fujishima, 1961), drake (Nishiyama et al., 1976) and turkey t o m (Fujihara and Nishiyama, 1984). Inspection of the cloacal region through a vaginal speculum revealed that the mucosal region in the vicinity of the papilla was red, presumably because it was rich in engorged blood vessles. This region referred to as TVP in this paper was probably the area producing a fluid containing aldose because it
73 gave a slight positive aldose c o l o r r e a c t i o n in s o m e birds in m a r k e d c o n t r a s t t o t h e negative r e a c t i o n b y fluid f r o m the o t h e r cloacal regions. T h e r e was a d i f f e r e n c e in the degree o f aldose c o l o r r e a c t i o n b e t w e e n t h e e j a c u l a t e d s e m e n and the fluid w h i c h o o z e d o u t f r o m the T V P o n t o a filter p a p e r o r seen w i t h t h e vaginal s p e c u l u m . This m a y be d u e t o the v o l u m e o f fluid t r a n s u d e d at e j a c u l a t i o n d u r i n g high sexual e x c i t e m e n t being greater t h a n was a p p a r e n t b y visual i n s p e c t i o n . T h e T V P in the guinea-fowl m a y be similar in h i s t o l o g y a n d f u n c t i o n t o t h e vascular tissue in t h e r o o s t e r ( L a k e and E1 J a c k , 1 9 6 6 ) , t h e E G R (ejacu l a t o r y g r o o v e region) in t h e d r a k e ( F u j i h a r a e t al., 1 9 7 6 ) and t h e T V P in t u r k e y t o m ( F u j i h a r a e t al., 1 9 8 5 a , b ) . T h e T V P , w h i c h is rich in b l o o d vessels, m a y be a special tissue c o m m o n t o male birds and a s o u r c e o f aldose in t h e c l o a c a d u r i n g ejaculation. T h e e j a c u l a t e d s e m e n o f t h e r o o s t e r (Nishiy a m a , 1 9 5 5 ; N i s h i y a m a a n d Fujishima, 1 9 6 1 ) , d r a k e ( N i s h i y a m a et al., 1 9 7 6 ) and t u r k e y t o m ( F u j i h a r a and N i s h i y a m a , 1 9 8 4 ) c o n t a i n s a b o u t 20% o f a c c e s s o r y r e p r o d u c t i v e fluids in the male t u r k e y and 50 p e r c e n t in the r o o s t e r and drake, d e p e n d i n g u p o n t h e m e t h o d o f collection. This c o u l d r e f l e c t t h e greater cloacal tissue swelling in t h e s e birds at ejaculation. T h u r s t o n e t al. ( 1 9 8 2 b ) m e n t i o n e d t h a t the seminal plasma a m i n o acids and p r o t e i n p r o f i l e s o f t h e guinea-fowl were similar t o t h o s e r e p o r t e d f o r t h e c h i c k e n and t u r k e y . T h e results d e s c r i b e d h e r e suggest t h a t male guinea-fowl a p p e a r t o have, in t h e cloaca, tissue similar t o t h e a c c e s s o r y r e p r o d u c t i v e organs in t h e t u r k e y t o m ( F u j i h a r a e t al., 1 9 8 5 a , b ) t h o u g h f u r t h e r e x p e r i m e n t s are n e e d e d t o s u b s t a n t i a t e the characteristics o f t h e s e m e n c o l l e c t e d b y n a t u r a l c o p u l a t i o n . ACKNOWLEDGEMENTS E x c e l l e n t t e c h n i c a l assistance was p r o v i d e d b y Mr. N. Y o s h i h i r o . REFERENCES Aire, T.A., Olowo-okorun, M.O. and Ayeni, J.S., 1980. The seminiferous epithelium in the guinea fowl (Numida meleagris). Cell Tissue Res., 205: 319--325. Ashizawa, K., Nishiyama, H. and Nagae, T., 1976. Effects of oviducal cells on the survival and fertilizing ability of fowl spermatozoa. J. Reprod. Fertil., 47: 305--311. Brillard, J.P. and Reviers, M., 1981. Testis development and daily sperm output in guineafowl raised under constant daily photoperiods. Reprod. Nutr, Develop., 21 (6B): 1105--1112. Burrows, W.H. and Quinn, J.P., 1937. The collection of spermatozoa from the domestic fowl and turkey. Poultry Sci., 16: 19--24. Fujihara, N. and Nishiyama, H., 1976a. Studies on the accessory reproductive organs in the drake. 3. Ejection mechanism of the fluid from the ejaculatory groove region. Poultry Sci., 55 : 1026--1030. Fujihara, N. and Nishiyama, H., 1976b. Studies on the accessory reproductive organs in the drake. 4. Effects of androgen on the ejaculatory groove region of the drake. Poultry Sci., 55: 1324--1330.
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