Sperm morphology in neotropical primates

The morphological and morphometric characterization of spermatozoa has been used as a taxonomic and phylogenetic tool for different species of mammals. We evaluated and compared the sperm morphometry of five neotropical primate species: Alouatta caraya, Ateles belzebuth and Ateles chamek of family Atelidae; and Cebus cay (=Sapajus cay) and Cebus nigritus (=Sapajus nigritus) of family Cebidae. After the collection of semen samples, the following parameters were measured on 100 spermatozoa from each specimen: Head Length, HeadWidth, Acrosome Length, Midpiece Length, MidpieceWidth and Tail Length. Considering the available literature on sperm morphometry, we gathered data of 75 individuals, from 20 species, 8 genera and 2 families. These data were superimposed on a phylogeny to infer the possible direction of evolutionary changes. Narrower and shorter spermatozoa seem to be the ancestral form for Cebidae, with a trend toward wider and larger heads in derived groups. The spermatozoa of Atelidae may show an increase in total length and midpiece length. Sperm heads would have become narrower in the more derived groups of Ateles. Sperm length may increase in the more derived species in both families. Our results are discussed in the context of sperm competition and sexual selection.Fil: Steinberg, Eliana Ruth. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución. Grupo de Investigación de Biología Evolutiva; ArgentinaFil: Sestelo, Adrián J.. Ecoparque Interactivo. Laboratorio de Biotecnología Reproductiva; ArgentinaFil: Ceballos, Maria Beatriz. Ecoparque Interactivo. Laboratorio de Biotecnología Reproductiva; ArgentinaFil: Wagner, Virginia. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución. Grupo de Investigación de Biología Evolutiva; ArgentinaFil: Palermo, Ana María. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución. Grupo de Investigación de Biología Evolutiva; ArgentinaFil: Mudry, Marta Dolores. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Ecología, Genética y Evolución. Grupo de Investigación de Biología Evolutiva; Argentin

Effect of thawing procedure on cryosurvival of deer spermatozoa: Work in progress

The objective of this study was to evaluate the effect of the thawing procedure on deer semen freezability. Frozen semen from the Genetic Resource Bank (GRB) of the Zoological Park of Buenos Aires (Argentina) was used. Seven mature stags (two red deer, two Père David’s deer and three fallow deer) were used as semen donors. Semen was diluted with a TRIS-egg yolk medium, packed in 0.25 ml straws and frozen in nitrogen vapour. For thawing, the frozen straws were subjected to the following procedures: (I) 70 °C, 5 s; (II) 50 °C, 8 s and (III) 37 °C, 10 s. Freeze-thaw motility percentage (FMP) and spermatozoa rating (FMR) were determined subjectively. Viability and acrosome integrity (NAR) were also assessed and the hypo-osmotic swelling test (HOST) was used to assess membrane integrity. Freeze-thaw motility percentage, FMR and NAR were assessed after an incubation of 1 h in citrate-yolk at 42 °C, and FMP and FMR after 2 h of incubation under the same conditions. The thawing procedure did not have an effect on the seminal characteristics evaluated immediately after this process. However, differences in FMP after 2 h of incubation (P<0.05) were found between the procedures, with the best overall recovery rates after freezing and thawing found with the use of protocols II (intermediate thawing) and III (slow thawing). Therefore, thawing protocols II and III, those that provide intermediate and slow thawing rates, were the most beneficial for semen thawing of the different cervid species analysed in this study.This study was sponsored by research funds of the Ministerio Español de Ciencia y Tecnologı́a (AGL2000-0671) and by Grant 190/PA-35 from the Consejerı́a de Agricultura y Medio Ambiente of the Junta de Comunidades de Castilla-La Mancha (Spain). The authors thank ALGAR SA for lending us the Père David’s deer stags, and Ma Dolores Pérez-Guzmán, for her help during the analysis of the data. Ana J. Soler is the recipient of a scholarship from the Consejerı́a de Ciencia y Tecnologı́a de la Junta de Comunidades de Castilla-La Mancha.Peer reviewe

Isolation of primary fibroblast culture from wildlife: the Panthera Onca case to preserve a South American endangered species

The Buenos Aires Zoo together with scientists from IByME-CONICET have created the Biobank of Genetic Resources (BGR) dedicated to the acquisition, processing, storage, use and supply of biomaterials to the scientific and conservation industry that are viable, diverse and representative of South American wildlife populations. Cell line establishment of somatic cells is a valuable resource to preserve genetic material of rare, difficult-to-find, endangered and giant species like Jaguar (Panthera onca), the largest South American felid. This work focuses on the isolation and culture of fibroblasts from Jaguar skin and muscle biopsies, and ear cartilage dissection immediately after death to preserve one of the several endangered species in this biome. These culture techniques enabled us to contribute 570 samples from 45 autochthonous and endangered species, including Jaguar. The fibroblasts obtained are a part of the 6700 samples, including tissues such as muscle, ovarian, testicular, blood, fibroblast cultures, sperm, hair, and fluids and cells from 450 individuals of 87 different species.Fil: Mestre Citrinovitz, Ana Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Sestelo, Adrián Jorge. Jardín Zoológico de la Ciudad de Buenos Aires S.A.; ArgentinaFil: Ceballos, María Belén. Jardín Zoológico de la Ciudad de Buenos Aires S.A.; ArgentinaFil: Barañao, Jose Lino Salvador. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Saragueta, Patricia Esther. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentin

Male reproductive traits, semen cryopreservation, and heterologous in vitro fertilization in the bobcat (Lynx rufus)

There is limited information on bobcat ejaculate traits and sperm cryopreservation and fertilizing ability. Bobcats were electroejaculated under general anesthesia in November (autumn) and April (spring), and endocrine and sperm traits were characterized. Testosterone (mean ± SEM: 0.90 ± 0.15 ng/mL) was not different between sampling times, but cortisol (average: 13.95 ± 1.73 μg/dL) was significantly higher in April. Average number of spermatozoa was 10.0 ± 3.4 × 106 sperm/ejaculate, with values being significantly higher in April. Sperm motility (average 55.7 ± 5.8% motile sperm) was not different between sampling times. The proportion of normal spermatozoa in the ejaculate (average: 14.7 ± 2.1%) was significantly higher in April, but the percentage of spermatozoa with intact acrosomes (average: 43.7 ± 3.8%) was significantly higher in autumn. Spermatozoa were cryopreserved in a Tes-Tris–based diluent (TEST) or Biladyl, both containing 20% egg yolk and 4% glycerol. Diluted sperm were loaded into straws, refrigerated using a programmable thermoblock with a dry chamber, frozen in nitrogen vapors, thawed, and incubated in F-10 medium with 5% fetal bovine serum for up to 3 h. After cryopreservation in TEST, there were about 50% motile sperm upon thawing, and survival was high during incubation post-thaw. Cryopreservation in Biladyl led to similar results, but motility decreased substantially during incubation post-thaw. Bobcat spermatozoa fertilized domestic cat oocytes matured in vitro. Fertilization rates were higher for sperm collected in April and cryopreserved in TEST (46%) than for those cryopreserved using Biladyl (<3%). Fertilized oocytes cleaved in culture, and some (27%) reached the morula stage. This study has allowed us to gain further baseline information on bobcat reproduction, explore sperm cryopreservation conditions, and show that fertilizing capacity can be tested using in vitro–matured cat oocytes. These results will be important for future conservation efforts.Natalia Gañán enjoyed scholarships from CSIC (I3P programme) and the BBVA Foundation. Raquel González enjoyed a scholarship from CSIC (I3P programme). Adrián Sestelo received a BBVA Foundation scholarship. E.R.S. Roldan is the recipient of a Royal Society Wolfson Research Merit Award. Funding from the Spanish Ministry of the Environment, CSIC, and the BBVA Foundation is gratefully acknowledged.Peer reviewe

Reproductive traits in captive and free-ranging males of the critically endangered Iberian lynx (Lynx pardinus)

et al.The Iberian lynx (Lynx pardinus) is the most endangered felid in the world. Adequate genetic management of in situ and ex situ populations, and linkage between both, require knowledge on male reproductive biology and factors influencing it. We examined the influence of age, free-ranging versus captive conditions and seasonality on phenotypic, endocrine and semen traits, and links between reproductive traits and male fertility. Males had relatively small testes, produced low sperm numbers, a low proportion of normal sperm, and a high proportion of motile sperm. Young (2-year-old) males had lower testosterone levels, fewer sperm, and a lower proportion of motile and normal sperm than R4-year-old males. No major differences were found in semen traits before and after the mating season or between free-ranging and captive males, although the latter had better sperm motility. Males with larger relative testes weight and more sperm copulated more frequently, whereas males that produced more sperm with higher motility produced more cubs per female. In conclusion, small relative testes size and low sperm quality could indicate either low levels of sperm competition or high levels of inbreeding. Young males are probably subfertile; there is a slight trend for males in the captive breeding programme to have better semen quality than wild males, and males with higher sperm production are sexually more active and more fertile. These findings have major implications for decisions regarding which males should breed, provide samples for the genetic resource bank, or participate in programmes involving the use of assisted reproductive techniques.Natalia Gañán received postgraduate support from the Spanish Research Council (CSIC-I3P programme) and the BBVA Foundation. Adrián Sestelo received a scholarship funded by the BBVA Foundation. E R S Roldan is the recipient of a Royal Society Wolfson Research Merit Award. This work was carried out with grants from the Spanish Ministry of the Environment (DG Biodiversity to E R S R, M G, and National Parks to F P), the Spanish Research Council (to E R S R and M G), the Ministry of Education and Science (to F P) and the BBVA Foundation (to M G).Peer reviewe