Effect of different semen extenders for the storage of chilled sperm in Tigrina (Leopardus tigrinus).

To enhance the conservation of endangered populations, the present study aimed to evaluate whether Tigrinas (Leopardus tigrinus) sperm could be conserved under refrigeration for short periods while maintaining sufficient quality for use in assisted-reproductive techniques (i.e., cryopreservation, in vitro fertilization). For this purpose, semen samples from 15 Tigrinas individuals were submitted to conventional and functional tests after different cooling periods (4 °C; 0, 12, and 24 hours postcooling), using TCM 199 (TCM), Ham's F10 (HAM), Ham's F10 with bovine serum albumin (HBSA), and Tris-citrate egg yolk (TEYC) extenders. In a second step, semen cooled using TEYC was supplemented with reduced glutathione (GSH) at different concentrations (0, 0.5, 1.0, and 1.5 mM). TEYC yielded superior results compared with TCM, HAM, and HBSA even after 24 hours of cooling in regard to the sperm motility index (SMI-TEYC: 50.2 ± 1.7%), high mitochondrial activity (TEYC: 51.4 ± 1.9%), plasma membrane integrity (TEYC: 53 ± 2.1%), and DNA integrity (TEYC: 56.3 ± 2.9%). In regard to the concentration of thiobarbituric-acid-reactive substances (TBARS), TEYC (1900.1 ± 341.4 ng/106 spermatozoa) showed higher levels compared with the other extenders (HAM: 638.7 ± 121.6 ng/106 spermatozoa; HBSA: 468.7 ± 95.6 ng/106 spermatozoa; TCM: 169.6 ± 31.6 ng/106 spermatozoa). However, GSH therapy had no effect. In conclusion, the TEYC extender may be useful in maintaining sperm parameters of Tigrinas for up to 24 hours at 4 °C. Furthermore, these results allow the transport of this material at a minimum quality to be further used for artificial insemination, in vitro fertilization, and the development of semen cryopreservation protocols.

Validation of non-fluorescent methods to reliably detect acrosomal and plasma membrane integrity of common marmoset (Callithrix jacchus) sperm.

Simple, rapid and stable sperm evaluation methods which have been optimized for common marmoset (Callithrix jacchus) are critical for studies involving collection and evaluation of sperm in the field. This is particularly important for new species groups such as Callitrichidae where the sperm have been little studied. Of this family, C. jacchus is the best known, and has been chosen as a model species for other members of the genus Callithrix. The fundamental evaluation parameters for sperm of any species are viability and acrosomal status. Semen samples were collected by penile vibratory stimulation. To evaluate sperm plasma membrane integrity, Eosin-Nigrosin was tested here for the common marmoset sperm to be used under field conditions. Further, a non-fluorescent stain for acrosome, the "Simple" stain, developed for domestic and wild cats, was tested on common marmoset sperm. This was compared with a fluorescent staining, Fluorescein isothiocyanate-Pisum sativum agglutinin (FITC-PSA), routinely used and validated for common marmoset at the German Primate Centre to evaluate acrosomal integrity. Results obtained with the "Simple" stain showed a marked differentiation between sperm with intact and non-intact acrosome both with and without ionophore treatment and closely correlated with results obtained with FITC-PSA. Temperature had no effect on the results with the "Simple" stain and the complete processing is simple enough to be carried out under field conditions. These findings indicated that the "Simple" stain and Eosin-Nigrosin provide rapid and accurate results for C. jacchus sperm and that those methods can be reliably used as field tools for sperm evaluation for this species.

Collection and evaluation of semen from captive howler monkeys (Alouatta caraya).

Semen samples (n=58) were collected by electroejaculation from nine adult male howler monkeys (Alouatta caraya) between November 2000 and August 2001 at the National Primates Center, Ananindeua, Brazil. The ejaculates were free of coagulum. Mean (+/-S.D.) values were: volume, 0.09 +/- 0.05 ml; pH, 8.1 +/- 0.5; concentration 649.5 +/- 926.7 x 10(6) sperm/ml; progressive motility, 75.8 +/- 18.1%; forward progressive sperm motility (scale, 0-5), 3.5 +/- 1.0; live spermatozoa, 68.3 +/- 15.0%; primary defects, 9.6 +/- 4.5%; and secondary defects, 11.8 +/- 4.6%. There were high correlations between motility and live sperm (r = 0.91, P < 0.01), motility and forward progressive sperm motility (r = 0.84, P < 0.01) and between forward progressive sperm motility and live sperm (r = 0.78, P < 0.01). There were no alterations observed during clinical examinations and hematological analysis performed before and after semen collection. Therefore, the method was considered safe and efficient. It can be used for the evaluation of the breeding potential of male howler monkeys in captivity and for the establishment of new assisted reproductive technology (ART) for threatened species of neotropical primates.

Bacterial septicemia in water snakes (Helicops modestus) in Brazil

Relata-se infecção septicêmica em cobras d’água (Helicops modestus) causada por Proteus vulgaris. Os animais iniciavam o processo apresentando abscessos puntiformes de coloração esbranquiçada na pele e após rápida evolução, morriam (100% de mortalidade). Descreve-se, ainda, a sensibilidade do P. vulgaris isolado dos animais a drogas antibacterianas.

Toxoplasmosis in emperor tamarin (Saguinus imperator): case report

A case of toxoplasmosis in an adult male emperor tamarin (Saguinus imperator) is reported. The primate was found dead and no clinical sign was noticed before death. Pathological findings included moderate to severe interstitial pneumonia, multifocal necrotizing hepatitis and multifocal to coalescing necrotizing lymphadenitis. Immunohistochemistry assays (strepto-avidin-biotin-peroxidase) performed on paraffin embedded tissues (lung, lymph nodes, liver, spleen, heart, intestine and adipose tissue) were strongly positive for Toxoplasma gondii

Colheita e avaliaçäo do sêmen em onça pintada (Panthera onca) / Semen collection and evaluation in the jaguar (Panthera onca)

Foram submetidas à eletroejaculaçäo 10 onças pintadas, de origem desconhecida, mantidas na Fundaçäo Parque Zoológico de Säo Paulo (n = 7) e Parque Zoológico Municipal "Quinzinho de Barros"- Sorocaba (n = 3), recebendo alimentaçäo de acordo com a dieta de rotina de cada instituiçäo. Nos 54 procedimentos realizados, foram obtidos ejaculados, ocorrendo contaminaçäo por urina em 3,7 por cento (n = 2) e em 11 (20,4 por cento) ocasiöes em que näo foram observados espematozóides. O volume médio obtido foi 7,42 ñ 3,69 ml, com motilidade média de 62,6 ñ 11,0 por cento e vigor de 2,71 ñ 0,52. A concentraçäo espermática foi 6,20 ñ 3,03 x 10 elevado a sexta potência espematozóides/ml, sendo que a porcentagem média de espermatozóides morfologicamente normais foi 46,7 ñ 5,8 por cento. A técnica de colheita de sêmen por eletroejaculaçäo é eficiente na espécie Panthera onça