Male reproductive cycle of Bothrops pubescens (Serpentes, Viperidae) from southern Brazil

We describe the seasonal variation in the male reproductive cycle of Bothrops pubescens, a pitviper from southern Brazil. All specimens were collected in the state of Rio Grande do Sul, a region with climates varying from subtropical to subtemperate and temperate. We characterized male reproduction considering histological analyses of the testes, sexual segment of the kidneys (SSK), and ductus deferens, as well as macroscopic indicators of reproductive activity (testes volume and ductus deferens width in the distal portion) to allow comparisons. We also evaluated the seasonal variation of epithelial height and tubule diameter of the testes and SSK. These reproductive parameters showed significant seasonal variation, with a high increase in summer and a peak in autumn for spermatogenic and SSK activity. Courtship behavior occurs on March (end of summer/beginning of autumn), in synchrony with sperm production and SSK hypertrophy. Sperm storage occurred in the ductus deferens throughout the year. We detected an ampulla ductus deferentis in the distal portion of the ductus deferens in some individuals. Male B. pubescens exhibit a seasonal reproductive cycle, and the use of microscopic methods was highly informative for supporting this conclusion.

Nós descrevemos a variação sazonal no ciclo reprodutivo de Bothrops pubescens, uma espécie de serpente da subfamília Crotalinae, da região Sul do Brasil. Os machos foram amostrados no estado do Rio Grande do Sul, que é uma região com climas variando de subtropical a subtemperado e temperado. Os ciclos dos testículos, segmento sexual renal (SSR) e ducto deferente foram caracterizados utilizando análises histológicas para descrever o ciclo reprodutivo dos machos. Nós também avaliamos indicadores macroscópicos da atividade reprodutiva (volume do testículo e largura do ducto deferente na região distal) para fins de comparação. Considerando os dados histológicos, foram avaliadas as variações sazonais da altura do epitélio e diâmetro dos túbulos tanto dos testículos quanto do SSR. Estes parâmetros reprodutivos apresentaram variação sazonal significativa, com grande aumento no verão e pico no outono, tanto para a atividade espermatogênica quanto para o SSR. Corte em vida livre ocorre em março (fim do verão/início do outono), em sincronia com a produção de esperma e a hipertrofia do SSR. Estocagem de esperma ocorreu no ducto deferente ao longo do ano. A ampulla ductus deferentis foi observada na porção distal do ducto deferente de alguns indivíduos. O ciclo reprodutivo dos machos de B. pubescens é sazonal e o uso de métodos microscópicos foi essencial para chegar a esta conclusão.

A histological and ultrastructural investigation of the female reproductive system of the water snake (Erythrolamprus miliaris): oviductal cycle and sperm storage

We studied the structural and cellular organisation of the oviduct of Erythrolamprus miliaris including its morphological variation during the reproductive cycle using light microscopy, scanning electron microscopy and transmission electron microscopy. Four anatomically distinct regions compose the oviduct of E. miliaris including the anterior and posterior infundibulum, glandular uterus, non-glandular uterus and pouch. The cells of the oviductal epithelium secrete material by apocrine and merocrine processes, which vary between the anatomical regions and according to each phase of the reproductive cycle. The infundibular epithelium secretes electron dense vacuoles, which suggests the production of lipids, whereas the epithelial secretion of the glandular uterus, non-glandular uterus and pouch creates lucent and slightly electron dense vacuoles, indicating the production of glycoproteins. The timing of mating, vitellogenesis and sperm storage directly influences the morphofunctional alterations in the oviducts of E. miliaris. Sperm storage occurs only in the infundibular receptacles with increased production of the neutral carbohydrates in the presence of male gametes. Sperm storage happens in vitellogenic, non-vitellogenic and pregnant females of E. miliaris. Thus, females may be able to produce multiple clutches at different seasons of the year regardless of mating during autumn.

Infundibular sperm storage and uterine muscular twisting in the Amazonian lancehead, bothrops atrox

Although knowledge about the reproduction of snakes of the genus Bothrops has increased recently, many questions remain regarding sperm storage by these neotropical pitvipers. The relationship between two known sperm storage sites—the nonglandular uterus and the infundibulum—has yet to be detailed for Bothrops. In addition, the formation of uterine muscular twisting (UMT) and its function in sperm storage in the nonglandular uterus have been questioned. This article presents data on both sperm storage sites (posterior infundibulum and nonglandular uterus) in the Amazonian lancehead, Bothrops atrox, aiming to address the role of UMT in bothropoid reproduction considering female reproductive stages, timing of sperm storage, and morphology and histochemistry of sperm receptacles. The oviduct of B. atrox is composed of the infundibulum (anterior and posterior), the glandular uterus, the nonglandular uterus and the pouch. Sperm storage occurs in the posterior infundibulum and nonglandular uterus in vitellogenic females. In the posterior infundibulum sperm is stored in receptacles, while in the nonglandular uterus it is stored in crypts in the UMT of vitellogenic females. Both sperm storage sites possess granules testing positive for neutral mucopolysaccharides in the presence of spermatozoa, possibly favoring the conditions for spermatozoa surveillance during sperm storage in the reproductive tract.

Infundibular sperm storage and uterine muscular twisting in the Amazonian lancehead, bothrops atrox

Although knowledge about the reproduction of snakes of the genus Bothrops has increased recently, many questions remain regarding sperm storage by these neotropical pitvipers. The relationship between two known sperm storage sites—the nonglandular uterus and the infundibulum—has yet to be detailed for Bothrops. In addition, the formation of uterine muscular twisting (UMT) and its function in sperm storage in the nonglandular uterus have been questioned. This article presents data on both sperm storage sites (posterior infundibulum and nonglandular uterus) in the Amazonian lancehead, Bothrops atrox, aiming to address the role of UMT in bothropoid reproduction considering female reproductive stages, timing of sperm storage, and morphology and histochemistry of sperm receptacles. The oviduct of B. atrox is composed of the infundibulum (anterior and posterior), the glandular uterus, the nonglandular uterus and the pouch. Sperm storage occurs in the posterior infundibulum and nonglandular uterus in vitellogenic females. In the posterior infundibulum sperm is stored in receptacles, while in the nonglandular uterus it is stored in crypts in the UMT of vitellogenic females. Both sperm storage sites possess granules testing positive for neutral mucopolysaccharides in the presence of spermatozoa, possibly favoring the conditions for spermatozoa surveillance during sperm storage in the reproductive tract.

A histological and ultrastructural investigation of the female reproductive system of the water snake (Erythrolamprus miliaris): oviductal cycle and sperm storage

We studied the structural and cellular organisation of the oviduct of Erythrolamprus miliaris including its morphological variation during the reproductive cycle using light microscopy, scanning electron microscopy and transmission electron microscopy. Four anatomically distinct regions compose the oviduct of E. miliaris including the anterior and posterior infundibulum, glandular uterus, non-glandular uterus and pouch. The cells of the oviductal epithelium secrete material by apocrine and merocrine processes, which vary between the anatomical regions and according to each phase of the reproductive cycle. The infundibular epithelium secretes electron dense vacuoles, which suggests the production of lipids, whereas the epithelial secretion of the glandular uterus, non-glandular uterus and pouch creates lucent and slightly electron dense vacuoles, indicating the production of glycoproteins. The timing of mating, vitellogenesis and sperm storage directly influences the morphofunctional alterations in the oviducts of E. miliaris. Sperm storage occurs only in the infundibular receptacles with increased production of the neutral carbohydrates in the presence of male gametes. Sperm storage happens in vitellogenic, non-vitellogenic and pregnant females of E. miliaris. Thus, females may be able to produce multiple clutches at different seasons of the year regardless of mating during autumn.

Mating plugs and male sperm storage in Bothrops cotiara (Serpentes, Viperidae)

We describe two new aspects of male reproduction in the pit viper Bothrops cotiara: the formation of mating plugs and epididymal sperm storage. Based on histological analyses of specimens (four males and three females), we propose that long-term sperm storage occurs in the ductus deferens, whereas macroscopic hypertrophy indicates that the epididymis may be a short-term sperm storage site. Secretions of the sexual segment of the kidneys probably form the mating plug that males deposit in the female vagina during copulation. A mating plug without sperm was observed in the spring. Future studies should address the functions and longevity of mating plugs and the role of the epididymis in Bothrops reproduction.

Reproductive biology of the brazilian lancehead, bothrops moojeni (Serpentes, Viperidae), from the state of Sao Paulo, Southeastern Brazil

Bothrops moojeni inhabits riparian forests and adjacent open, wet areas in the Brazilian Cerrado. We address several aspects of the reproductive biology of a population from southeastern Brazil, including sexual maturation and dimorphism, male and female reproductive cycles, and sperm storage. We also provide the first record of male-male combat behavior in the wild. We sampled specimens held in the scientific collection of the Butantan Institute, Sao Paulo, Brazil. After macroscopic analyses and dissections, we collected the testes of males and the posterior uterus of females showing uterine muscular twisting for microscopic analyses. Males < 54.5 cm and females < 58.0 cm were sexually immature. Mature females are larger than males, although male-male combat is reported for this species. Male-male fighting and copulation occur during late summer in the wild. Histological analysis of the testes confirmed that sperm production occurs simultaneously with those reproductive events. Females showed an extended period of vitellogenesis, from February to November (late summer to spring). We observed stored sperm in the female reproductive tract during autumn. Because mating is not synchronous with ovulation, sperm storage is an obligatory component of the reproductive cycle of B. moojeni. Ovulation begins in August (winter) but most females with the presence of embryos in the oviducts occurred from January to March (summer). Parturition in captivity occurred in February, confirming that births are concentrated in the summer. Given that the Cerrado is highly fragmented in southeastern Brazil, studies on the reproductive biology of additional B. moojeni populations might provide useful information regarding intraspecific variation in reproduction in pit vipers.