Cryopreservation of collared peccary (Pecari tajacu) semen using different freezing curves, straw sizes, and thawing rates

AbstractThe objective of this study was to verify the effect of different freezing curves, straw sizes, and thawing rates on the cryopreservation of collared peccary semen. Twelve ejaculates were obtained from captive adult males by electroejaculation, and evaluated for sperm motility, kinetic rating, viability, morphology, and functional membrane integrity. The ejaculates were diluted in a coconut water extender (ACP-116c) with egg yolk and glycerol, packaged into 0.25mL or 0.50mL plastic straws and cryopreserved in liquid nitrogen following a slow (−10°C/min) or a fast (−40°C/min) freezing curve. After one week, samples were thawed at 37°C/1min or 70°C/8s and evaluated as reported for fresh semen, and also for kinematic parameters (computerized analysis). A significant decrease in sperm motility and kinetic rating was observed after glycerol addition at 5°C and also after thawing for all the treatments (P<0.05). Regarding post-thaw semen variables, no differences were verified between freezing curves when the same straw size and thawing rate were taken as reference (P>0.05). In general, values for sperm characteristics found after thawing at 37°C were better preserved than at 70°C (P<0.05), both in the use of 0.25mL or 0.50mL straws, which were similar for semen packaging (P>0.05). The evaluation of the kinematic parameters of sperm motility confirmed these results at values varying from 20% to 30% motile sperm for the samples thawed at 37°C, and values fewer than 12% motile sperm for samples thawed at 70°C (P<0.05). In conclusion, we recommend the use of a fast freezing curve that reduces the time spent on the cryopreservation of collared peccary semen, which could be packaged both in 0.25mL or 0.50mL straws, but the thawing should be conducted at 37°C/1min

Simple-sequence repeat markers of Cattleya coccinea (Orchidaceae), an endangered species of the Brazilian Atlantic Forest

Microsatellite markers were developed for the endangered Brazilian orchid species Cattleya coccinea to describe its genetic diversity and structure and to support conservation studies. Nine microsatellite loci were isolated and characterized using an enriched genomic library. All loci are polymorphic at least in the 2 populations sampled, except for loci Cac05 and Cac09 for the Petropolis population. the mean number of alleles per locus was 8.8 between populations. the mean values of the observed and expected heterozygosities were 0.541 (ranging from 0 to 1) and 0.639 (ranging from 0 to 0.9), respectively. Cross-amplifications were performed in 7 additional Epidendroideae species, and at least 2 loci were successful in 3 additional Cattleya species, Epidendrum secundum, and Brasiliorchis gracilis. All markers described herein will be useful in further studies evaluating the genetic diversity, population dynamics, and conservation genetics of C. coccinea and related species

Establishing the hypoosmotic swelling test for sperm analysis in collared peccaries (Pecari tajacu)

Soluções hiposmóticas com diferentes concentrações (0, 50, 100, 150, 200mOsm/L) foram testadas para a avaliação funcional da membrana espermática de catetos (n=13). Foi verificado que o número de espermatozoides reagidos diminuía (P<0,05) de acordo com o aumento da osmolaridade do meio. A maior porcentagem (71,8%) de espermatozoides reagidos, bem como a menor variação nas respostas osmóticas, foi detectada com o uso de água destilada (0mOsm/L) (P<0,05), a qual também apresentou a menor variação nos resultados, de acordo com os erros padrão verificados. Em conclusão, a água destilada aparenta ser uma solução adequada para o uso no teste hiposmótico para sêmen de catetos

Morphoagronomic genetic diversity in american wild rice species

To characterize the genetic variability among species and populations of South American wild rice, eleven populations of Oryza glumaepatula, seven of O. grandiglumis, four of O. latifolia and one of O. alta, from Brazil and Argentina, were evaluated. A greenhouse experiment was conducted in completely randomized blocks with 23 treatments. Twenty morphoagronomic traits were assessed. Univariate analyses were performed with 16 quantitative traits with the partitioning of populations within species. Significant differences (p<0.001) between species were observed for all the traits as well as among populations within the species. The most variable was O. glumaepatula followed by O. latifolia. Multivariate discriminant canonical and cluster analyses confirmed the separation of the highly diverse O. glumaepatula populations from the tetraploid species, and the high genetic variation among O. latifolia populations. Morphological differences among the three tetraploid species seemed to be enough to ascribe them at least the condition of species in statu nascendi.<br>Visando caracterizar a diversidade genética entre espécies e populações de arroz selvagem da América do Sul, foram avaliadas 11 populações de Oryza glumaepatula, sete de O. grandiglumis, quatro de O. latifolia e uma população de O. alta, originárias do Brasil e Argentina. Foi conduzido um experimento em casa-de-vegetação em blocos ao acaso com 23 tratamentos. Vinte caracteres agro-morfológicos foram avaliados. Análises univariadas foram realizadas para 16 caracteres quantitativos, desdobrando-se o efeito de populações dentro de espécies. Diferenças significativas (p<0,001) entre espécies foram observadas para todos os caracteres bem como entre populações dentro de espécies. A mais variável foi O. glumaepatula seguida de O. latifolia. Análises de agrupamento e discriminante canônica confirmaram a separação das populações de O. glumaepatula das espécies tetraplóides, e a grande variação genética entre populações de O. latifolia. Diferenças morfológicas entre as três espécies tetraplóides parecem suficientes para classificá-las como espécies pelo menos na condição statu nascendi

Morphology and vascularization of the corpus luteum of peccaries (Pecari tajacu, Linnaeus, 1758) throughout the estrous cycle

The current paper characterizes the changes in morphology and vascularization of the corpus luteum of collared peccaries during the estrous cycle and correlates progesterone synthesis (P4). Twenty females were subjected to a treatment for estrus synchronization; an ear implant containing 1.5 mg of norgestomet was implanted on D0, whereas on D9 the peccaries received an IM injection of eCG 200UI and 50g of PGF2a. The animals were divided into four groups (G1, G2, G3 and G4) and euthanized on post-ovulation days 3, 12, 18 and 22. The ovaries were collected and the corpora lutea were measured and processed for histological and vascular density (Dv). Blood was collected for dosage of P4 serum. The morphology of the ovaries, the corpora lutea and P4 varied significantly during the estrous cycle (P<0.001). There was a significant co-relationship between weight and length of the ovaries and CL (r = 0.66, r = 0.52, P<0.05, respectively) and between weight, length and width of the CL and P4 (r = 0.51, r = 0.54 and r = 0.68, P<0.05, respectively). The luteal Dv was highly influenced by the estrous cycle phase (P<0.0001). The P4 and luteal Dv concentrations were higher in G2 and evidenced maximum secretory activity, with a highly significant correlation (P<0.0001). Assessed lutein parameters may estimate the phase of the estrous cycle in peccaries and the functional activity of the corpus luteum

Indução do estro em cutias (Dasyprocta leporina) utilizando-se protocolos à base de prostaglandina isolada ou em associação com análogo de GnRH

RESUMO Comparou-se a eficiência de protocolos para indução de estro em cutias. Em cinco fêmeas, foram administradas duas doses de cloprostenol (5µg) com intervalo de nove dias, via intraperitoneal; em outras cinco, administraram-se 30µg de análogo do hormônio liberador de gonadotrofinas (GnRH), via intravulvar, seguidos de 5µg de cloprostenol, via intraperitoneal, após sete dias e, após mais dois dias, nova dose do análogo de GnRH. A cada três dias, a ciclicidade reprodutiva dos animais foi monitorada, por meio de coleta de sangue, para dosagem hormonal, ultrassonografia ovariana e citologia vaginal. Duas das fêmeas que receberam apenas prostaglandina, as quais estavam em fase luteal no início do tratamento, manifestaram o estro aos três e seis dias após a segunda administração da droga. Já nas fêmeas que receberam a prostaglandina associada ao análogo do GnRH, duas que originalmente estavam em fase luteal apresentaram estro aos quatro dias após o tratamento, e uma outra apenas após 10 dias. Não foram evidenciadas diferenças estatísticas quanto à eficiência dos tratamentos (P>0,05). Conclui-se que, de acordo com os protocolos utilizados, o uso da prostaglandina isolada ou em associação com análogo do GnRH para a indução do estro em cutias D. leporina apresenta eficiência limitada às fêmeas que estejam em fase luteal por ocasião do início do tratamento

Allelic Diversity In Populations Of Solanum Lycocarpum A. St.-hil (solanaceae) In A Protected Area And A Disturbed Environment [diversidade Alélica Em Populações De Solanum Lycocarpum A. St.-hil (solanaceae) Em Unidade De Conservação E Em Ambiente Sob Influência Antrópica]

This study aimed to compare the genetic diversity of populations of Solanum lycocarpum A.St.-Hil between natural and human disturbed environments, with the assumption that protected areas have greater genetic diversity than disturbed areas. For this study, two populations were sampled in Goiás State, Brazil. One was located in a conservation unit, Serra de Caldas Novas State Park, in the Caldas Novas municipality. The other was located in a pasture area in the municipality of Morrinhos. The two populations are 41 km apart. We sampled 60 individuals from each population, which were genotyped with five microsatellite loci (SSR). The highest number of alleles was recorded in the population of the conservation unit, where we found 11 exclusive and five rare alleles. In the disturbed area, we recorded only three exclusive alleles and one rare allele. Although we did not observe significant inbreeding in these populations, genetic divergence between them was high (G ST (Hedrick)=0.147 =0.147) for a species with long distance seed dispersal. The results corroborate the hypothesis that the population in the less disturbed area harbors greater allelic diversity. They also confirm the effectiveness of using protected areas to preserve the genetic diversity of the species.254937940Buso, G.S.C., Brondani, R.V., Amaral, Z.P.S., Reis, A.M.M., Ferreira, M.E., (2000) Desenvolvimento De Primers SSR Para Análise Genética Depimentas E Pimentões (Capsicum Spp.) Utlizando Biblioteca Genômica Enriquecida, p. 15. , Brasília, Embrapa/Cenargen. (Boletim de pesquisaCouternay, Y., Conservation of the Maned Wolf: Fruitful relations in a changing environment (1994) Canid News, 2, pp. 41-43Crow, J.F., Aoki, K., Group selection for polygenic behavioral trait: Estimating the degree of population subdivision. Proceedings of the National Academy of Sciences of United States of América (1984) PNAS, 81 (19), pp. 607-677Degen, B., (2006) Genetic Data Analysis and Numerical Tests, , GDA_NT 2006, Grosshansdorf, Institute for Forest GeneticsFerreira, M.E., Grattapaglia, D., (1988) Introdução Ao Uso De Marcadores RAPD E RFLP Em Análise Genética, , 3 ed. Brasília, Embrapa - CenargenGoodman, S.J., RstCalc: A collection of computer programs for cal- culating estimates to genetic differentiation from microsatellite data and determining their significance (1997) Molecular Ecology, 6 (9), pp. 881-885Hedrick, P.W., A standardized genetic differentiation measure (2005) Evolu- Tion, 59 (8), pp. 1633-1638Lewis, P.O., Zaykin, D., (2000) Genetic Data Analysis: Computer Program For the Analysis of Allelic Data, , http://lewis.eeb.uconn.edu/lewishome/software.html, Version 1.0 (d15). Free program dis- tributed by the author over the internet from the GDA., Acesso em 23/01/2007Martins, K., Chaves, L.J., Buso, G.S.C., Kageyama, P.Y., Mating system and fine-scale spatial genetic structure of Solanum lycocarpum St. Hill. (Solanaceae) in the Brazilian Cerrado (2006) Conservation Genetics, 7 (6), pp. 957-969Martins, K., Chaves, L.J., Vencovski, R., Kageyama, P.Y., Genetic structure based on nuclear and chloroplast microsatellite loci of Sola- num lycocarpum A. St.-Hil (Solanaceae) in Central Brazil (2011) Genetics and Molecular Research, 10 (2), pp. 665-677Moura, T.M., Sebbenn, A.M., Chaves, L.J., Coelho, A.S.G., Oliveira, G.C.X., Kageyama, P.Y., Diversidade e estrutura genética espacial em populações fragmentadas de Solanum spp. do Cerrado, estimados por meio de locos microssatélites (2009) Scientia Forestalis, 37 (82), pp. 143-150Oliveira, F.A.T., Oliveira, L.C.A., Biologia floral de uma população de S. lycocarpum St. Hill. (Solanaceae) em Lavras MG (1988) Revista Brasileira De Botânica, 11 (1-2), pp. 23-32Vidal, M.C., Stacciarini-Seraphin, E., Camara, H.H.L.L., Crescimento de plântulas de Solanum lycocarpum St. Hil. (lobeira) em casa de vegetação (1999) Acta Botanica Brasilica, 13 (3), pp. 271-275Weir, B.S., Cockerham, C.C., Estimating F-statistics of the analysis of populations structure (1984) Evolution, 38 (6), pp. 1358-137