Comparison of semen characteristics and sperm cryopreservation in common marmoset (Callithrix jacchus) and black-tufted-ear marmoset (Callithrix penicillata).

BACKGROUND: The common marmoset (Callithrix jacchus) is one of the most studied Neotropical primates regarding reproduction. However, little has been reported on the black-tufted-ear marmoset (Callithrix penicillata), which may produce fertile hybrids with other Callithrix. This is the first study to evaluate sperm freezing from black-tufted-ear marmoset. METHODS: Testicles from all animals were measured, and semen was collected by penile vibrostimulation. Samples were analyzed after collection, after chilling and addition of glycerol, and after thawing. RESULTS AND CONCLUSIONS: Fresh semen from both species was similar in many aspects. Additionally, there is a relationship between total motility, plasma membrane integrity, and acrosome integrity observed in sperm samples from both species. We managed to evaluate fresh and thawed sperm with suitable methods for use under zoo or field conditions. However, TEST egg yolk with glycerol at 4% and 6% concentration was not effective for sperm protection in both species during cryopreservation.

Testicular volume and semen characteristics in the endangered southern muriqui (Brachyteles arachnoides).

BACKGROUND: The southern muriqui (Brachyteles arachnoides) is an endangered Neotropical primate. Semen collection and description of its traits, as well as testicular morphometry, have never been reported for this species. METHODS: Testicles from five healthy adult captive southern muriqui were measured, and semen was collected by rectal probe electrostimulation (RPE). RESULTS AND CONCLUSIONS: A solid coagulum was identified in all ejaculates, and none of them liquefied, spontaneously or non-spontaneously. It was possible to collect semen using RPE, and although solids coagula did not liquefy, we managed to describe ejaculates characteristics and also confirmed that southern muriqui have relatively large testes size. Further investigations are needed to improve coagulum handling, to achieve a better spermatozoa recovery aiming its application in assisted reproductive technologies.

Assessment of different sperm functional tests in golden-headed lion tamarins (Leontopithecus chrysomelas).

The golden-headed lion tamarin (Leontopithecus chrysomelas) is an endangered species endemic to Brazil's Atlantic Forest, a shrinking biodiversity hotspot. As in other Neotropical primates, its semen characteristics and freezability are poorly studied. Hence, reproductive technologies for callitrichids would greatly benefit from reliable methods of semen analysis. In a bid to promote reproductive research in tamarins, we validated simple and inexpensive sperm function tests that can be used to monitor sperm-egg binding, plasma membrane and acrosome integrity, mitochondrial activity, and DNA fragmentation. Ejaculates from adult males were individually diluted and divided into control and damage-induced aliquots, and then samples comprising assorted amounts of damaged spermatozoa were examined by organelle-specific tests. Our findings showed that sperm-binding in chicken egg perivitelline membrane (EPM) positively correlated with the number of spermatozoa injured by snap-freezing. Eosin-nigrosin (EN) and propidium iodide readings were correlated with each other, and both provided robust measurements of plasma membrane integrity. A high correlation between expected and measured amounts of acrosome-intact spermatozoa was found using Fast Green-Rose Bengal (FG-RB), Coomassie Blue (CB), and FITC-PSA stains, and all three methods exhibited comparable results. Likewise, different percentages of UV-irradiated spermatozoa were accurately assessed for DNA integrity by Toluidine Blue (TB) and sperm chromatin dispersion (SCD) tests. Comparisons between 3,3'-diaminobenzidine (DAB) and JC-1 stains also indicated the reliability of the former assay to ascertain gradual increases in spermatozoa with greater mitochondrial function. These data confirmed that different parts of the tamarin spermatozoa can be simply and consistently evaluated by EPM, EN, FG-RB, CB, TB, and DAB protocols.

Semen cryopreservation in golden-headed lion tamarin, Leontopithecus chrysomelas.

Wild animal genetic resource banking (GRB) represents a valuable tool in conservation breeding programs, particularly in cases involving endangered species such as the golden-headed lion tamarin (Leontopithecus chrysomelas). Thus, we aimed to assess a sperm freezing protocol for golden-headed lion tamarins using two different exenders: BotuBOV® (BB) and Test Yolk Buffer® (TYB). Ejaculates were collected by penile vibrostimulation from animals housed at São Paulo Zoological Park Foundation, São Paulo, Brazil, and after immediate analysis, two aliquots were diluted in BB and TYB. Postthawing samples were evaluated for total and progressive motility, plasma membrane and acrosome integrities, mitochondrial activity, susceptibility to oxidative stress, and sperm-egg-binding. No differences between BB and TYB were found for most seminal parameters, except for acrosome integrity and susceptibility to oxidative stress (in both cases BB showed higher values). However, in spite of these differences and regardless of the extender used, postthaw sperm motility and viability with the described protocol were encouraging (on average >50% and >80%, respectively), indicating that sperm cryopreservation may be a short-term measure for the conservation of golden-headed lion tamarins.

Collection, Evaluation, and Coagulum Dissolution of Semen from Goeldi's Monkey, Callimico goeldii.

Research on Neotropical primates' reproduction is necessary due to the lack of available information and the increasing threat to these species. Callimico goeldii is listed as Vulnerable on the IUCN Red List of Threatened Species. This study aimed to test rectal electrostimulation for semen collection and evaluate seminal characteristics. Therefore, semen from 6 captive Goeldi's monkeys was collected and, for the first time, seminal characteristics are described. Coagulum formation was noted in all ejaculates, and we obtained partial or complete liquefaction of the samples. Results were (means ± SD): volume = 26.9 ± 11.87 µL; pH = 7.61 ± 0.28; concentration = 143.18 ± 174.96 × 106 spermatozoa/mL; total sperm motility = 83.33 ± 5.16%; linear progressive motility = 46 ± 24.08%; plasma membrane integrity = 36.38 ± 16.11%; acrosome integrity using fast-green/bengal-rose staining = 63.41 ± 11.72%, and kit Spermac® = 69.36 ± 11.81%; abnormal sperm = 72.5 ± 17.7%, with 16.2 ± 7.7% major defects and 56.3 ± 10% minor defects; sperm with high mitochondrial activity class I = 16.45 ± 22.25%. Rectal electrostimulation was an efficient method for semen collection in this species. Investigations are required to improve semen collection and handling, including cryopreservation methods.

Seasonal effects on testes size and sustained semen quality in captive golden-headed lion tamarins, Leontopithecus chrysomelas.

The golden-headed lion tamarin (Leontopithecus chrysomelas) is an endangered primate that, according to timing of births, there seems to have seasonal reproductive patterns in captive populations inhabiting the Southern Hemisphere. The extent to which male tamarins have circannual cyclic fluctuations in testicular functions, however, remains unknown. Changes in testis size and seminal variables, therefore, were evaluated during the rainy and dry seasons of the year in seven males. There was estimation of mating and birth seasons from the breeding colony using a 27 year database (229 birth records). Births were distributed throughout the year with peaks occurring in August-October and January-March (39.7 % and 30.5 % of all births, respectively). Semen collection using penile vibrostimulation was successfully induced in more than 96 % of the attempts regardless of the season (total of 75 ejaculates). Body mass did not vary significantly between seasons, but relative testes size was larger during the dry season. Values for none of the seminal variables (total sperm count, total and progressive motility, plasma membrane and acrosome integrity, and total sperm defects), however, were different during the rainy and dry seasons. These results indicate that testicular function in golden-headed lion tamarins may not be affected by daylength changes, and that seasonal patterns of female reproduction is perhaps more relevant for the reproductive timing of the species. Furthermore, the possibility of year-round production of ejaculates containing viable sperm broadens our perspective of preserving genetic diversity within the species because there is a greater opportunity for semen collection and freezing.