Effect of photoperiod on characteristics of semen obtained by electroejaculation in stump-tailed macaques (Macaca arctoides)

Some environmental variables determining seasonal reproduction in mammals are temperature, humidity, food availability, and photoperiod. Among these, photoperiod is considered the main regulator of primates’ seasonal reproduction, thus the latitudinal distribution of primate populations is a key factor determining the appearance of seasonal reproduction. The present work presents supporting discrete seasonality in male stump-tailed macaques (Macaca arctoides). We investigated whether semen quality and testosterone covaried with Mexico City’s photoperiod and relative humidity by analyzing variations in the portions that form the ejaculate: the seminal liquid, the seminal coagulum, and the copulatory plug. Five male adult stump-tailed macaques were electroejaculated once a month, obtaining three semen samples per male, from August 2011 to July 2012 (except for December 2011) (n = 165). Our results showed that stump-tailed macaque sperm counts were significantly different between the portions of the ejaculate. The seminal coagulum contained the significantly largest number of spermatozoids, followed by the copulatory plug and the seminal fluid. Photoperiod and relative humidity had major influence on the sperm count in the seminal coagulum and the testosterone concentrations. Testosterone reached its highest values around the time when days and nights lasted the same hours, decreasing when days either grew longer or became shorter. Concerning relative humidity, sperm counts in the seminal coagulum were highly variable on dry days, but decreased as the relative humidity increased. We conclude that stump-tailed macaques have a discrete seasonality, occurring in spring and fall when macaques’ reproductive condition and readiness for postcopulatory intrasexual competition increase.     

Highly Polymorphic Microsatellite Markers for the Assessment of Male Reproductive Skew and Genetic Variation in Critically Endangered Crested Macaques (<Emphasis Type="Italic">Macaca nigra</Emphasis>)

Genetic analyses based on noninvasively collected samples have become an important tool for evolutionary biology and conservation. Crested macaques (Macaca nigra), endemic to Sulawesi, Indonesia, are important for our understanding of primate evolution as Sulawesi macaques represent an exceptional example of primate adaptive radiation. Crested macaques are also Critically Endangered. However, to date we know very little about their genetics. The aim of our study was to find and validate microsatellite markers useful for evolutionary, conservation, and other genetic studies on wild crested macaques. Using fecal samples of 176 wild macaques living in the Tangkoko Reserve, Sulawesi, we identified 12 polymorphic microsatellite loci through cross-species polymerase chain reaction amplification with later modification of some of these primers. We tested their suitability by investigating and exploring patterns of paternity, observed heterozygosity, and evidence for inbreeding. We assigned paternity to 63 of 65 infants with high confidence. Among cases with solved paternity, we found no evidence of extragroup paternity and natal breeding. We found a relatively steep male reproductive skew B index of 0.330 ± 0.267; mean ± SD) and mean alpha paternity of 65% per year with large variation across groups and years (29–100%). Finally, we detected an excess in observed heterozygosity and no evidence of inbreeding across our three study groups, with an observed heterozygosity of 0.766 ± 0.059 and expected heterozygosity of 0.708 ± 0.059, and an inbreeding coefficient of ?0.082 ± 0.035. Our results indicate that the selected markers are useful for genetic studies on wild crested macaques, and possibly also on other Sulawesi and closely related macaques. They further suggest that the Tangkoko population of crested macaques is still genetically variable despite its small size, isolation, and the species’ reproductive patterns. This gives us hope that other endangered primate species living in small, isolated populations may also retain a healthy gene pool, at least in the short term.     

Male fertility in natural populations of red deer is determined by sperm velocity and the proportion of normal spermatozoa

Male reproductive success is determined by the ability of males to gain sexual access to females and by their ability to fertilize ova. Among polygynous mammals, males differ markedly in their reproductive success, and a great deal of effort has been made to understand how selective forces have shaped traits that enhance male competitiveness both before and after copulation (i.e., sperm competition). However, the possibility that males also may differ in their fertility has been ignored under the assumption that male infertility is rare in natural populations because selection against it is likely to be strong. In the present study, we examined which semen traits correlate with male fertility in natural populations of Iberian red deer (Cervus elaphus hispanicus). We found no trade-offs between semen traits. Our analyses revealed strong associations between sperm production and sperm swimming velocity, sperm motility and proportion of morphologically normal spermatozoa, and sperm viability and acrosome integrity. These last two variables had the lowest coefficients of variation, suggesting that these traits have stabilized at high values and are unlikely to be related to fitness. In a fertility trial, our results show a large degree of variation in male fertility, and differences in fertility were determined mainly by sperm swimming velocity and by the proportion of morphologically normal sperm. We conclude that male fertility varies substantially in natural populations of Iberian red deer and that, when sperm numbers are equal, it is determined mainly by sperm swimming velocity and sperm morphology.     

Male facial attractiveness and masculinity may provide sex‐ and culture‐independent cues to semen quality

Phenotype‐linked fertility hypothesis (PLFH) predicts that male secondary sexual traits reveal honest information about male fertilization ability. However, PLFH has rarely been studied in humans. The aim of the present study was to test PLFH in humans and to investigate whether potential ability to select fertile partners is independent of sex or cultural background. We found that on the contrary to the hypothesis, facial masculinity was negatively associated with semen quality. As increased levels of testosterone have been demonstrated to impair sperm production, this finding may indicate a trade‐off between investments in secondary sexual signalling (i.e. facial masculinity) and fertility or status‐dependent differences in investments in semen quality. In both sexes and nationalities (Spanish and Colombian), ranked male facial attractiveness predicted male semen quality. However, Spanish males and females estimated facial images generally more attractive (gave higher ranks) than Colombian raters, and in both nationalities, males gave higher ranks than females. This suggests that male facial cues may provide culture‐ and sex‐independent information about male fertility. However, our results also indicate that humans may be more sensitive to facial attractiveness cues within their own populations and also that males may generally overestimate the attractiveness of other men to females.     

Effect of preservation of human semen sample at 4–6 and 25 °C on sperm motility

Sperm motility is the result of transverse movements that exist along its tail. It plays an important role in male fertility. The aim of this study was to evaluate the influence of keeping washed normozoospermic semen samples at 4–6 and 25 °C on the motility of spermatozoa. 26 semen samples of normozoospermic were washed twice in modified Ham’s F10 medium. Then, thirteen of the semen samples were kept in refrigerator (4–6 °C) and the remaining samples were stored in incubator (25 °C) for 12 days. On the 0 (immediately after sampling as control group), 1st, 2nd, 5th, 7th and the 12th days, the percentage of fast progressive (grade a), slow progressive (grade b), non-progressive (grade c) and immotile (grade d) sperm cells were calculated for each temperature. The data obtained from this study showed that the percentages of a, b and c grades of motile spermatozoa were significantly decreased (p?<?0.001) during 12 days at the both temperatures but reduction of these percentages has a gentle slope at 4–6 °C. There was no motile sperm after 12 days of storage. This study suggests that motile spermatozoa could be retrieved up to 7 days after the storage of washed normozoospermic men semen samples at 4–6 and 25 °C. Also, there were no motile sperm cells 12 days after sampling.     

Patterns of infection by intestinal parasites in sympatric howler monkey (Alouatta palliata) and spider monkey (Ateles geoffroyi) populations in a tropical dry forest in Costa Rica

In primate populations, endoparasite species richness and prevalence are associated with host traits such as reproductive and social status, age, sex, host population density, and environmental factors such as humidity. We analyzed the species richness and prevalence of intestinal parasites in two sympatric primate populations, one of Alouatta palliata and one of Ateles geoffroyi, found in a tropical dry forest in Costa Rica. We identified three species of intestinal parasites (Controrchis sp., Trypanoxyuris sp., and Strongyloides sp.) in these two primate species. We did not find any differences in species richness between the primate species. However, the prevalences of Controrchis sp. and Trypanoxyuris sp. were higher in Alouatta palliata. Similarly, males and lactating females of Alouatta palliata showed higher Controrchis sp. prevalences. We did not observe any differences in parasite richness and prevalence between seasons. Infectious diseases in endangered primate populations must be considered in conservation strategies, especially when defining protected areas.     

The use of nonmetal electrodes in electroejaculation of restrained but unanesthetized macaques

A successful technique for electroejaculation with nonmetallic electrodes cut from defibrillation pads is described. Twenty-six adult male cynomolgus and eleven adult male rhesus macaques were electroejaculated while immobilized with chair restraint. From 123 attempted electroejaculations in both species of macaques, 119 semen specimens were obtained. The volume, concentration, % motility, and % normal forms of cynomolgus and rhesus macaque semen are presented. The use of nonmetallic electrodes provides a high quality ejaculate while minimizing the risks of adverse affects on valuable populations of macaques.     

The breeding management affects fresh and cryopreserved semen characteristics in Melopsittacus undulatus

Melopsittacus undulatus is a companion parrot worldwide diffused. Many parrots are considered endangered or vulnerable. The preservation of semen is crucial in endangered species, thus, M. undulatus could be a good model to study sperm characteristics and semen cryopreservation in these other endangered parrots. In this study the effect of the breeding management (males bred in promiscuous aviary or in couple) on sperm characteristics (motility, membrane integrity and morphometry) of fresh and cryopreserved semen was evaluated. The computer-assisted sperm analysis (CASA) revealed a significant effect of the husbandry method on semen characteristics in budgerigars: male housed in couple with the female in individual cages allowed the higher results in term of both semen quantity and sperm quality. Total and progressive motility were significantly higher in males bred in couple (68.7 ± 8.9% and 54 ± 15.9%, respectively) than in promiscuous aviary (48.3 ± 15.1% and 24.4 ± 12.4%, respectively), such as sperm velocity (average path velocity, straight line velocity, and curvilinear velocity). The type of sperm movement (amplitude of lateral head displacement, beat cross frequency, straightness, and linearity), sperm membrane integrity and morphometry parameters seemed not affected by the husbandry method. The standardization of a CASA procedure for the semen analysis in M. undulatus allow further studies on parrot semen manipulation and cryopreservation, but the method used for the breeding of the male could have a significant effect on the semen quality.     

Intraspecies variation in dominance style of Macaca fuscata

Knowledge of intraspecific variation is important to test the evolutionary basis of covariation in primate social systems, yet few reports have focused on it, even in the best-studied species of the Macaca genus. We conducted a comparative study of the dominance styles among three provisioned, free-ranging groups of Japanese macaques at Shodoshima Island, Takasakiyama Mountain and Shiga Heights, and collected standard data on aggressive and affiliative behavior during a period of 5 years. Our data in the Takasakiyama and Shiga groups support previous studies showing that Japanese macaques typically have despotic social relations; nevertheless, our data in the Shodoshima group are inconsistent with the norm. The social traits of Shodoshima monkeys suggested that: (1) their dominance style is neither despotic nor tolerant but is intermediate between the two traits; (2) some measures of dominance style, e.g., frequency and duration of social interactions, covary as a set of tolerant traits in Shodoshima monkeys. This study suggests broad intraspecific variation of dominance style in Japanese macaques as can be seen in some other primate species.     

Fertility assessment of cheetah males with poor quality semen

Reports on semen quality of the cheetah (Acinonyx jubatus) indicate that high percentages of abnormal morphs and sperm concentrations, 10 times lower than in domestic cats, are found in all populations. These characteristics are believed to result from unusual genetic homozygosity, hypothesized to have been caused by passage of the species through one or more population bottlenecks during its recent history. In a sample of 12 captive-living males, we found semen characteristics to be equal or inferior to those previously reported for males living in other captive facilities. Ten of these males (83.3%) nevertheless produced pregnancies. Seventeen of 19 pregnancies resulted from matings during a single estrus. This examination of the reproductive potential of males having comparatively inferior ejaculate quality supports the suggestion that husbandry programs may be more significant than physiological impairment in causing the low birth rates of captive cheetahs. These results also have implications for ascertaining fertility thresholds in mammalian populations undergoing increased levels of inbreeding as a consequence of habitat deterioration. © 1993 Wiley-Liss, Inc.     

Effect of Dietary Selenium and Vitamin E on Ganders’ Response to Semen Collection and Ejaculate Characteristics

Compared to other domestic bird species, geese exhibit the lowest reproductive efficiency (poor semen quality, low egg production, and poor fertility and hatchability rates). From an economic perspective, it is a necessity of improve these reproductive traits. Studies have demonstrated that the essential trace element—selenium—plays key roles in testicular development and the maintenance of spermatogenesis. The aim of the present study was to determine the effect of feed supplementation with organic selenium and vitamin E on ganders’ response to manual semen collection and semen quality. Sixteen 3-year-old White Koluda ganders were randomly divided into two groups. The control group was provided commercial feed while the experimental group was provided with the same commercial feed supplemented with selenium (0.3 mg/kg) and vitamin E (100 mg/kg). The response of individual ganders from both groups to manual semen collection and the quality of the semen collected were evaluated. The supplements increased (P?≤?0.05) the frequency and decreased the time interval of a complete ejaculatory response of the ganders to manual semen collections (82.7 % supplement vs. 73.5 % control). Males from the supplemented group had significantly higher (P?≤?0.01; P?≤?0.05) ejaculate volumes, sperm concentrations, and percentages of viable sperm and lower percentages of immature sperm (spermatids). Lipids peroxidation, expressed in terms of the malondialdehyde concentration, was lower (P?≤?0.01) in semen of the supplemented group (0.172 nmol/50?×?10⁶) as compared to the controls (0.320 nmol/50?×?10⁶). Moreover, the duration of the reproductive period of the ganders in the experimental group was 1 week longer. The results show that supplemental dietary selenium and vitamin E improved both the ganders’ response to manual semen collection and semen quality. We conclude that such feed supplementation could lead to greater economic benefits through increased reproductive efficiency within the goose production industry.     

Apyrene sperm from the triploid donors restore fecundity of cryopreserved semen in Bombyx mori

Female moths of Bombyx mori were artificially inseminated with cryopreserved semen. The fertility of inseminated females varied from 0% to 76.9% depending on the strain. Addition of fresh semen from triploid males, which are infertile but whose semen includes intact apyrene sperm, greatly improved fecundity of cryopreserved semen from normal males. Frozen apyrene sperm from the triploid donors also improved the fecundity of females, inseminated with cryopreserved normal semen, but less than fresh semen from triploid males. Fertilization success in B. mori requires the presence of both, intact eupyrene and apyrene sperm. Our results show that eupyrene sperm tolerate the cryopreservation process better than apyrene sperm. Hence, we recommend to add apyrene sperm from the triploid donors as helper sperm routinely to cryopreserved semen in artificial insemination. This may advance the application of cryopreservation as a routine technique to maintain silkworm resources. The technique may also be applicable to other moth and butterfly species which, like B. mori, possess eupyrene and apyrene sperm.     

Meiotic behavior and pollen fertility in triploid and tetraploid natural populations of Campuloclinium macrocephalum (Eupatorieae,Asteraceae)

Campuloclinium macrocephalum DC. is a perennial herb widely distributed in the New World and introduced in South Africa, where it is commonly called “pompom weed”. This species is considered one of the most important weeds of Brazil and one of the problematic invasive plants of South Africa. The meiotic system can be studied to assess the ability of a weed to spread, but only few studies on C. macrocephalum have been realized. In this study, we examined the meiotic behavior and pollen fertility of 14 natural populations of C. macrocephalum from Argentina and Uruguay. Meiotic analysis revealed 2 triploid (2n = 3x = 30), 11 tetraploid (2n = 4x = 40) and 1 mixed population (2n = 2x = 20, 2n = 4x = 40). Both, triploid and tetraploid specimens showed a widely variable meiotic behavior with irregular chromosome pairing showing univalents, bivalents, trivalents (in triploids) and tetravalents (in tetraploids) at diacinesis of first meiotic division. Different abnormalities were observed, such as: laggard chromosomes, chromatin bridges, and out of plate chromosomes at metaphase I. During meiosis I (prophase), some cells showed the phenomenon of cytomixis or chromatin transfer between pollen mother cells. The meiotic indexes suggest that only four populations were normally fertile (over 90 % of fertile pollen), indicating meiotically stable plants. The remaining populations share variable pollen fertility, with triploids ranging from 46.64 to 54.83 % and tetraploids varying from 3.54 to 45.30 %. We suggest that polyploidy seems to be recurrent in C. macrocephalum, promoting partial sterility of pollen grains, generating large numbers of individuals by apomixis promoting invasion of crop fields. This study presents the meiotic behavior of this weed, these could be useful for future studies of biological control in areas with no natural enemies.     

Male-to-male differences in post-thaw motility of rhesus spermatozoa after cryopreservation of replicate ejaculates

BACKGROUND: The efficiency of controlled propagation to produce rhesus monkeys of particular genotypes can be maximized by use of cryopreserved spermatozoa collected from specific males to inseminate appropriate females. But this assumes that semen from males with different genotypes can be cryopreserved with equal effectiveness. METHODS: To investigate whether spermatozoa from different Macaca mulatta males can be effectively cryopreserved when frozen under identical conditions, we collected and froze semen specimens from 13 adult, fertile males maintained at three primate research centers. RESULTS: Survival, based on post-thaw motility normalized to the pre-freeze value, was assayed within 30 minutes after thawing; it varied from 50% to 70% but declined thereafter. To examine the response of semen from individual males, we collected and froze three to six ejaculates per male from each of seven males. CONCLUSIONS: In general, semen from a given male responded reproducibly to freezing, but there were significant differences among males. The cause of these differences among M. mulatta males in post-thaw sperm survival remains unidentified.     

Fertility of undiluted ram epididymal spermatozoa stored for several days at 4°C

In vitro preservation of the male gamete is a challenge in the development of artificial insemination techniques for domestic animals. Specific strategies and diluents have been developed for the preservation of the fertilizing ability of the semen for each species. However, the epididymal medium has been demonstrated to be the best sperm environment to maintain sperm viability over several days and weeks for mammals. The aims of this study were to evaluate the motility and in vivo fertility of ram epididymal spermatozoa when the semen was stored for up to 4 days at 4°C undiluted in epididymal plasma. The study was undertaken with two ovine breeds (Ile de France and Corriedale). The motility of epididymal spermatozoa was better preserved in the undiluted epididymal fluid than when epididymal spermatozoa were diluted in classic ovine extender such as skim milk. During storage, the decrease in the percentage of motile sperm was lower if the epididymal spermatozoa were collected immediately after epididymal sampling than 24 h after castration or animal death. The fertility obtained after cryopreservation of the stored sperm and subsequent intrauterine insemination ranged from 55% to 24% following 24 to 96-h sperm storage. There was a linear regression relationship between fertility and the number of motile sperm inseminated for both breeds. These results show that it is possible to keep epididymal sperm motile and fertile for several days without dilution. Such a method of sperm preservation could be a final possibility for animals of high genetic value or for endangered species when the collection of semen before death of the animal is not possible.     

Preservation of Turkey spermatozoa by the use of emulsions and supercooling methods

Techniques are described for the emulsification and supercooling of turkey spermatozoa. Selection of an appropriate emulsification medium was carried out after tests for spermicidal properties and droplet stability were performed. Differential thermal analysis (DTA) studies were employed to measure the temperatures of spontaneous nucleation of the suspended semen droplets during cooling.During the fertility trial it was shown that the entrapment of spermatozoa in Dow 230 carrying fluid reduced the fertility (P < 0.05) from 91.6 to 81.2%. Additional significant reductions were observed for 1 hr at ?21 °C (64.9%) and during the separation of the carrying fluid and semen via centrifugation (57.2%). This indicates that the separation of the Dow 230 from the semen is not necessary and the semen can be artificially inseminated in its emulsified form.The decline in fertility of the emulsified semen stored at ?21 °C for 1 hr could be attributed to possible damage during freezing of some suspended semen droplets as seen in thermograms of emulsified semen. Additional work in this area should be directed toward elucidating the controlling factors of supercooling as they relate to possible in vitro cryopreservation.     

Predation of artificial nests by introduced rhesus macaques (<Emphasis Type="Italic">Macaca mulatta</Emphasis>) in Florida,USA

Native throughout Asia, rhesus macaques are believed to have the widest native range of any non-human primate and are capable of adapting to an extensive diversity of habitats. Rhesus macaques have caused environmental degradation in introduced habitats, including decreasing bird populations through nest predation. In the 1930s, rhesus macaques were intentionally introduced into what is today Silver Springs State Park (SSSP), central Florida, in an effort to increase tourism. Our objective was to determine whether introduced rhesus macaques in SSSP would consume eggs presented in artificial nests. We used camera traps adjacent to 100 open-cupped artificial bird nests baited with quail eggs near the Silver River. Nests were placed in shrubs and left in the field site for 12 days, representative of the incubation period of native passerine species. Twenty-one nests were depredated by rhesus macaques, nine by nest predators other than macaques, and five nests by an unidentified predator. Nests were more likely to be depredated by macaques when located in areas of high macaque relative abundance. This study suggests introduced rhesus macaques may influence nest predation rates of native bird species in natural areas.     

Genotyping from semen of wild Japanese macaques (Macaca fuscata)

The noninvasive collection of animal cells is crucial for DNA analyses in wild populations that cannot be disturbed by capture. We describe the collection of 68 semen samples following copulation and masturbation events in wild habituated and nonhabituated troops of Japanese macaques on the protected island of Yakushima. We used this DNA to amplify 390 base pairs (bp) of the mitochondrial DNA control region in 16 individuals from eight troops, and found a monomorphic pattern in agreement with the low variability imposed by geographic isolation and female philopatry. We also amplified two microsatellite loci from samples collected after the resident males of a focal troop had copulated with different females. We found several different allele combinations in samples collected after the observed mating of a single male, indicating the presence of contaminant DNA, presumably from males that had previously mated with the same female. This discovery made it impossible to assign a given sample to a specific male except when the samples were recovered after masturbation events. Thus, it was not possible to test for kinship or estimate allele frequencies from the semen samples. The mixing of semen, and the pattern of sample collection observed in morphologically identified individuals support the notion that strong mating and sperm competition exists among resident and nonresident males.     

Male display rate reveals ejaculate characteristics in the Trinidadian guppy,Poecilia reticulata.

Previous work (Reynolds and Gross 1992) has demonstrated that mate choice enables female guppies, Poecilia reticulata, to obtain heritable benefits (''good genes'') for their offspring. It is not yet known whether males also signal their functional fertility, that is, their viability as fathers, to female guppies. Our study tested the hypothesis that a male''s behaviour reveals characteristics of his ejaculate. We uncovered a strong correlation between display rate and sperm number in male guppies derived from two wild Trinidadian populations. This result indicates that information on the functional fertility of potential mates is available to females. However, our investigation also found a significant relationship between the frequency of sneaky mating and sperm number. Female guppies in wild populations, particularly those experiencing a high predation regime, are constantly subjected to sneaky mating attempts. As the males responsible for these sneaky matings have highest reserves of sperm they may have the capacity to undermine female choice. The implications of these findings for sperm competition are discussed.     

Progesterone levels in seasonally breeding,free-ranging male <Emphasis Type="Italic">Macaca thibetana</Emphasis>

Progesterone, a “female” hormone, modulates sexual behaviors in male mammals. This modulation is well documented in laboratory animals, but it is as yet unknown whether progesterone is associated with testosterone and/or sexual behaviors in primates living in their natural environments. In this study, we collected a total of 426 fecal samples and approximately 453 h of behavioral data from five male Tibetan macaques (Macaca thibetana) to study relationships between males’ progesterone and testosterone levels and sexual behaviors. Our results showed a negative correlation between fecal testosterone and progesterone levels. Both hormones seasonally varied. For three of the five subjects, we also found that copulatory and sexually motivated behaviors were negatively correlated with fecal progesterone levels. While not significant for the other two males, this study provides insight into the relationship between progesterone and the sexual behaviors of male primates living in a natural environment.