Post-testicular maturation of spermatozoa is crucial for attaining the morphological and functional capabilities needed for successful fertilization. Epididymal epithelia offer a favorable environment for spermatozoa that are stored long term in the turtle epididymis; however, sperm–epithelial interactions during storage, which are enormously important for sperm functional and morphological maturation, are still largely unknown in turtles. The present study examined the epididymis during the sperm-storage period (November–April) in the Chinese soft-shelled turtle (Pelodiscus sinensis). Light and transmission electron microscopy were used to determine the cellular features of each epididymal segment (caput, corpus, and cauda) and their epithelial interactions with the spermatozoa. Spermatozoa were mainly located in the lumena of caput, corpus, and cauda epididymides. Numerous spermatozoa were bound to apical surfaces of the epithelia, and several were even embedded in the epithelial cytoplasm of the caput and corpus epididymides. No embedded spermatozoa were found in the cauda epididymis. In all epididymal segments, principal and clear cells showed the synthetic activity, evidenced by a well-developed endoplasmic reticulum network and high and low electron-dense secretory materials, respectively. Principal and clear cells in the caput and corpus segments showed embedded spermatozoa in electron-dense secretions and in the lipid droplets within the cytoplasm. No lysosomes were observed around the embedded spermatozoa. The lumena of the caput and corpus segments showed few apocrine and low electron density secretions. In the lumen of the cauda epididymidis, different secretions, such as holocrine with low and high electron density and their fragmentation, apocrine, and dictyosome, were found and are summarized. Altogether, sperm physical interactions with secretions either in the cytoplasm of epithelium or in the lumen may support the viability, morphological maintenance, and transfer of various proteins involved in long-term sperm storage in the turtle. This interaction could help us to understand the mechanisms of long-term sperm storage and provide more insights into the reproductive strategies of turtle sperm preservation.

In birds, after natural mating or artificial insemination (AI), sperms are transported through cloaca or vagina at the utero-vaginal junction (UVJ) of oviduct. Only 1% of the deposited sperm passes through the selection process in the vagina to reach the UVJ. In the UVJ, sperm enter the tubular invagination of the surface epithelium of the mucosa collectively called ‘sperm storage tubules’ (SSTs) where they are stored for longer duration retaining their fertilising capacity. The storage capacity and duration varies from species to species. This enables the birds to produce fertile eggs continuously. Shortly before and during egg production, sperm residing in the SST upon release will ascend the oviduct to the site of fertilisation in the infundibulum. Within the SSTs, sperm motility is suppressed and is initiated at some stage before fertilisation. Restoration of sperm motility is necessary for the release of spermatozoa from the SSTs. The exact mechanism by which sperm are stored in these tubules and released has not been established and hence, this is the focus of this review.

Female crayfish stores male gametes after mating until the beginning of egg laying and fertilization. The aim of the present study was to investigate the duration of post-mating spermatophore storage as well as the timing and temperature of spawning in two crayfish species of economic importance, namely the signal crayfish Pacifastacus leniusculus and the noble crayfish Astacus astacus. Results showed that the average duration of the post-mating spermatophore storage is significantly (P<0.05) longer in the noble crayfish (34.6±1.7 days, range: 19 to 60 days) than the signal crayfish (3.9±0.5 days, range: 1 to 18 days). The highest percentages of the post-mating spermatophore storage duration in the signal crayfish (46.5%) and the noble crayfish (44.5%) were 1 and 31 to 40 days, respectively. While there is an overlap in the timings of mating and egg laying in the signal crayfish, these two reproductive processes were not observed at the same days in the noble crayfish and there was at least 2 weeks interval between last mating and first egg laying individuals. Average mating and egg laying temperatures were significantly (P<0.05) higher in the signal crayfish than the noble crayfish. The average temperatures for mating in both species were significantly (P<0.05) higher than the temperatures that they utilized for egg laying. In conclusion, female noble crayfish stores post-mating spermatophores a longer duration compared with the signal crayfish. Also, the signal crayfish mates and lays egg in temperatures that are higher than the noble crayfish. Spawning season is shorter in the signal crayfish compared with the noble crayfish. The results of present study provide information contributing to the crayfish broodstock management in aquaculture.

The sterile insect technique (SIT) has been proposed as an area-wide method to control the South American fruit fly, Anastrepha fraterculus (Wiedemann). This technique requires sterilization, a procedure that affects, along with other factors, the ability of males to modulate female sexual receptivity after copulation. Numerous pre-release treatments have been proposed to counteract the detrimental effects of irradiation, rearing and handling and increase SIT effectiveness. These include treating newly emerged males with a juvenile hormone mimic (methoprene) or supplying protein to the male's diet to accelerate sexual maturation prior to release. Here, we examine how male irradiation, methoprene treatment and protein intake affect remating behavior and the amount of sperm stored in inseminated females. In field cage experiments, we found that irradiated laboratory males were equally able to modulate female remating behavior as fertile wild males. However, females mated with 6-day-old, methoprene-treated males remated more and sooner than females mated with naturally matured males, either sterile or wild. Protein intake by males was not sufficient to overcome reduced ability of methoprene-treated males to induce refractory periods in females as lengthy as those induced by wild and naturally matured males. The amount of sperm stored by females was not affected by male irradiation, methoprene treatment or protein intake. This finding revealed that factors in addition to sperm volume intervene in regulating female receptivity after copulation. Implications for SIT are discussed.

Squids of the family Bathyteuthidae have generally been infrequently encountered in their deep-sea habitat. Remotely operated vehicles were used to observe seven individuals in situ in the Monterey Submarine Canyon, CA, USA. One of these was a female Bathyteuthis berryi holding a sheet with approximately 360 embedded embryos. Examination of this female after collection revealed the presence of a seminal receptacle on the buccal membrane. We present some potential costs and benefits of post-spawning egg care, a strategy that is now known for two families of deep-dwelling squids, but may turn out to be more common with the increasing exploration of the deep sea.

Earwigs (Insecta, Dermaptera) show astonishing diversity in penis morphology. In several families, males have a single penis (termed virga), whereas males of other families possess two functional virgae. Taxonomists have assumed that the two-virgae state is ancestral; however, ecological reasons why the ancestor acquired two virgae have not been explored. This study investigated in detail male and female genital structures, mating behaviour and insemination processes in the earwig Diplatys flavicollis (Diplatyidae). Diplatyidae are considered to be the most primitive family of earwigs. SEM and light-microscopy revealed that males of this species have two gonopores on each of two virgae, similar to those reported in other diplatyids (i.e. two double-barrelled penises), while females have four to six independent sperm-storage organs (spermathecae). Rapid fixation of mating pairs and insemination success of males from which one virga had been removed clearly revealed that only one virga was used for mating and was usually sufficient for inseminating multiple spermathecae. This finding rejects the one-to-one correspondence between male gonopores and female spermathecae. Based on allometric analysis of spermathecal variation, the possible significance of multiple spermathecae in relation to sperm-storage strategies of females is discussed. Compared to studies of male genital morphology, few studies have described spermathecal morphology. Based on compiled data of spermathecal and virgal morphology among earwigs, a parallel evolutionary trend between spermathecae and virgae from complicated (multiple) to simple (single) ones is suggested, and further investigation of the phylogeny, female genital morphology and insemination processes among earwigs is encouraged.

Birds, reptiles and insects (e.g. Hymenoptera) share fundamental similarities in their reproductive strategies due to the presence, in the female tract, of specialized sites in which spermatozoa may reside for prolonged periods upon a single mating. In avian species, two distinct storage sites are present, one located in the utero-vaginal junction and the other in the lower portion of the infundibulum. At both sites, spermatozoa are stored in sperm storage tubules (SSTs) which are discrete, generally non-branched invaginations of the luminal epithelium (see review by Bakst et al., 1994). The SSTs located in the uterovaginal junction are considered as the main site of residence of spermatozoa upon their deposition in the lower portion of the vagina. The fertilizing potential of females along the reproductive season is at first dependent on their ability to store and maintain adequate populations of spermatozoa in their SST in order to repeatedly provide the site of fertilization with sufficient numbers of “fit” spermatozoa after each ovulation. As a consequence in poultry species, eggs from females with prolonged sperm storage potential have also a natural tendency to maintain optimal fertility rates for prolonged periods. This situation may be highly desirable to sustain high fertility in the case of partial failure of the males during the season.

This review is an attempt to address the main intrinsic and extrinsic factors capable of favouring or altering fertility in poultry species. Unfortunately, most of the information available is obtained from two species, namely the domestic fowl and turkey, which, in certain circumstances, may not reveal enough to pinpoint the controlling factors of fertility in other poultry species.

The functioning of female genitalia in many arthropods, especially the site and mode of sperm storage, may have a strong impact on the pattern of sperm usage, leading to differential male fertilization success and even varying offspring sex ratios. The female genital morphology of the linyphiid spider Pityohyphantes phrygianus was investigated, as this species was shown to possess an intriguing mechanism of sex ratio determination, possibly related to genital morphology. By means of SEM and serial semi-thin sectioning, two distinctly different spermathecae plus an additional sperm sac, equipped with several valves, were found on each side of the genital apparatus. Sperm is stored under different conditions, as one of the spermathecae exhibits large gland cells of different types that discharge their products into the spermatheca through long secretory ducts, whereas the other spermatheca is surrounded by a simpler type of gland cell. Furthermore, there are folds instead of closed ducts leading to the spermathecae, and from the spermathecae to the oviduct. The latter fertilization-fold runs inside the copulatory chamber and enters the oviduct from the outside. The mechanisms for sex ratio bias in spiders are discussed. The occurrence of two types of spermatheca of which one is twisted, provides good evidence for the involvement of female genital morphology in sex ratio control of P. phrygianus offspring. Data are compiled on species for which information is available on sex ratio and female genital morphology. We put forward a hypothesis that in species with a skewed sex ratio, there is interaction between female genital morphology and behaviour, i.e. change of body position before, during or immediately after sperm transfer.

The morphology of the sperm storage organ (spermatheca) may influence the outcome of sperm competition in a variety of invertebrate species. Simultaneously hermaphroditic pulmonates show considerable inter- and intraspecific variation in the structure of the spermatheca. The morphological variation in the spermatheca (number of spermathecal tubules and tubule length) and the amount of sperm stored in the tubules in 115 individuals of the simultaneously hermaphroditic land snail Arianta arbustorum (L.) from six natural populations in the Eastern Alps (Gesäuse, Austria) was examined. The populations differed in snail density, therefore in the number of potential mating partners (range: 0.9–39.8 individuals/m2) and thus in the sperm competition risk. The number of spermathecal tubules ranged from two to nine. However, the six populations did not differ in either the mean number of spermathecal tubules, the length of the main tubule (range 0.9–3.4 mm), or in the cumulative length of the tubules (range 1.5–7.8 mm). The number of tubules and tubule length were not correlated with either shell height and shell breadth or with population density. The amount of sperm stored in the spermathecal tubules was estimated. Individuals from different populations did not differ in the amount of sperm stored, and the amount of sperm stored was not correlated with population density. Results indicate that the risk of sperm competition does not affect the complexity of the spermatheca (number of spermathecal tubules) in this snail. However, the possibility that individuals in high-density populations store sperm from more different mating partners than those in low-density populations remains to be examined.

The male reproductive cycles of three species of vespertilionid bat (Nyctophilus geoffroyi, N. major and Vespadelus regulus) are described. Descriptions are based on the degree of testicular and cauda epididymal development estimated from field captures, and for N. geoffroyi, on histological examination of fixed specimens and assays of their testicular testosterone concentrations. All species were found to undergo a dissociated pattern of reproduction; the activity of the testes and accessory glands of reproduction were asynchronous. Spermatogenesis occurred in summer/early autumn (December–March) and testes generally regressed before mating began in autumn (March–May). Spermatozoa were stored in the cauda epididymides during the mating period. Leydig cells of N. geoffroyi involuted prior to mating but the prostate gland remained enlarged and secretory during winter (June–August), regressing in late spring (November). Testicular testosterone concentrations were greatest when Leydig cells were maximally enlarged, and concentrations were low during the mating period. The reproductive cycle of all species apparently involved asynchrony between primary and secondary sexual function, which is typical of vespertilionid bats inhabiting temperate latitudes. As mating occurs when sperm stores cannot be replenished, selection for prudent sperm allocation may be expected.