Biotecnologias na preservação de espermatozoides felinos / Biotechnologies in the preservation of feline sperm

O gato doméstico é a única espécie da família Felídea sem risco ou iminência de extinção, diferente da maior parte dos felinos selvagens. Desta forma, o desenvolvimento e aprimoramento de diferentes biotécnicas reprodutivas, são essenciais para a manutenção da qualidade reprodutiva, tendo em vista a preservação de espécies mais vulneráveis. Além disso, as biotécnicas do sêmen são para as tecnologias reprodutivas, como a inseminação artificial (IA) e a fertilização in vitro (FIV). Sendo assim, o objetivo deste compilado bibliográfico foi abordar as principais técnicas de colheita, análise e preservação de sêmen/espermatozoides felino, assim como o uso dessas células em IA e FIV. Para a colheita do sêmen felino, diferentes métodos têm sido aplicados: ejaculação farmacológica, eletroejaculação e vagina artificial. Em caso de óbito do reprodutor, os espermatozoides recuperados do epidídimo também apresentam viabilidade reprodutiva. Ademais, a cinética espermática avaliada pelo sistema CASA, a morfologia e a morfometria são as principais análises que demonstram a qualidade espermática e refletem na fertilidade do ejaculado. O sistema CASA também avalia a trajetória individual de cada espermatozoide, que ao se agrupar em clusters, demonstra a heterogeneidade do ejaculado nas subpopulações. Contudo, os diluentes para a conservação e refrigeração dos espermatozoides felinos e as curvas de congelação ainda não estão totalmente estabelecidos e influenciam diretamente a viabilidade dos espermatozoides criopreservados. Diante disso, os resultados da utilização do sêmen felino após criopreservação são inconsistentes, sendo necessários mais estudos para elucidar melhores curvas de congelação e meios de diluentes para viabilizar a preservação do material genético dos gatos.

The domestic cat is the only species of the Felidea family without risk or imminence of extinction, unlike most wild cats. Therefore, the development and improvement of different reproductive biotechnologies are essential for the maintenance of reproductive quality for the preservation of the most vulnerable species. Furthermore, semen biotechnologies are the basis for reproductive technologies such as artificial insemination (AI) and in vitro fertilization (IVF). Thus, the objective of this bibliographic compilation was to approach the main techniques of collection, analysis, and preservation of feline semen/sperm, as well as the use of these cells in AI and IVF. For feline semen collection, different methods have been applied: pharmacological ejaculation, electroejaculation, and artificial vagina. In case of death of the sire, sperm recovered from the epididymis also show reproductive viability. Moreover, the sperm kinetics evaluated by the CASA system, the morphology, and the morphometry are the main analyzes that demonstrate sperm quality and reflect on ejaculate fertility. The CASA system also evaluates the individual path of each sperm, which, when grouped into clusters, demonstrates the heterogeneity of the ejaculate in the subpopulations. However, diluents for the conservation and refrigeration of feline sperm and freezing curves are not yet fully established and directly influence the viability of cryopreserved sperm. Therefore, the results of using feline semen after cryopreservation are inconsistent, and further studies are needed to elucidate better freezing curves and diluents to enable the preservation of the genetic material of cats.

Protocolos para contenção química e eletroejaculação para coleta de sêmen de queixadas (Tayassu pecari) / Chemical restraining and electroejaculation protocols for semen collection in white-lipped peccary (Tayassu pecari)

Background: In order to reverse the White-lipped peccary decline, besides protecting its habitat and controlling hunting,it is necessary a captive breeding program. There are reports, however, on the low fertility of white-lipped peccary, makingit difficult its reproduction in captivity, making artificial insemination one of the main tools to prevent the loss of geneticdiversity of species kept in captivity. Information on safe methods of anesthesia and the collection of semen should beinvestigated. Therefore, we aimed to compare the effects of the anesthetic protocols acepromazine/ketamine and xylazine/ketamine, as well as electroejaculation protocols, for semen collection in white-lipped peccary (Tayassu pecari).Materials, Methods & Results: Twelve adult male white-lipped peccaries were submitted both to the xylazine/ketamineand acepromazine/ketamine anesthetic protocols. The anesthetic induction time and duration, the degree of muscle relaxation, the time for anesthetic recovery and the quality of the animals’ recovery were evaluated. Additionally, the qualityof the sedation was evaluated based on the animal’s behavior. We also evaluated the effect of drugs on erectile functionsas well as the efficiency of 3 electroejaculation protocols with increasing or fixed voltages (2 to 4 V; 5 to 12 V; 12 V). Theacepromazine/ketamine combination promotes shorter induction time, duration and recovery from anesthesia than thexylazine/ketamine association. There were no differences, however, between the tested anesthetic protocols in relation toheart rate, respiratory rate and temperature. Ejaculate was obtained from only 2 animals when using the xylazine/ketamineprotocol and adoption of stimuli between 5 and 12 V, with 10 stimuli at each voltage. In turn, ejaculate was obtained from4 animals submitted to the acepromazine/ketamine protocol, 3 of them with the adoption of stimuli between 5 and 12 V...

Protocolos para contenção química e eletroejaculação para coleta de sêmen de queixadas (Tayassu pecari) / Chemical restraining and electroejaculation protocols for semen collection in white-lipped peccary (Tayassu pecari)

Background: In order to reverse the White-lipped peccary decline, besides protecting its habitat and controlling hunting,it is necessary a captive breeding program. There are reports, however, on the low fertility of white-lipped peccary, makingit difficult its reproduction in captivity, making artificial insemination one of the main tools to prevent the loss of geneticdiversity of species kept in captivity. Information on safe methods of anesthesia and the collection of semen should beinvestigated. Therefore, we aimed to compare the effects of the anesthetic protocols acepromazine/ketamine and xylazine/ketamine, as well as electroejaculation protocols, for semen collection in white-lipped peccary (Tayassu pecari).Materials, Methods & Results: Twelve adult male white-lipped peccaries were submitted both to the xylazine/ketamineand acepromazine/ketamine anesthetic protocols. The anesthetic induction time and duration, the degree of muscle relaxation, the time for anesthetic recovery and the quality of the animals recovery were evaluated. Additionally, the qualityof the sedation was evaluated based on the animals behavior. We also evaluated the effect of drugs on erectile functionsas well as the efficiency of 3 electroejaculation protocols with increasing or fixed voltages (2 to 4 V; 5 to 12 V; 12 V). Theacepromazine/ketamine combination promotes shorter induction time, duration and recovery from anesthesia than thexylazine/ketamine association. There were no differences, however, between the tested anesthetic protocols in relation toheart rate, respiratory rate and temperature. Ejaculate was obtained from only 2 animals when using the xylazine/ketamineprotocol and adoption of stimuli between 5 and 12 V, with 10 stimuli at each voltage. In turn, ejaculate was obtained from4 animals submitted to the acepromazine/ketamine protocol, 3 of them with the adoption of stimuli between 5 and 12 V...(AU)

Características do comportamento hiperativado de espermatozoides de felinos domésticos / Characteristics of the cat sperm hyperactivated behavior

Sperm hyperactivity is a physiological behavior, and in the feline species it is characterized by an increase in the curvilinear velocity (VCL) and a greater head beat (ALH) evaluated by the CASA system. The study aimed to compare and correlate kinetic parameters of Hyperactive and Non-Hyperactive feline ejaculates when considering the means of VCL and ALH. Ejaculates were collected by electro ejaculation from 21 cats. The seminal samples had the kinetic parameters evaluated by the CASA system. From the average values of VCL and ALH, the ejaculates were classified in group HP (Hyperactivated, when VCL>190.17µm/s and ALH>6.44µm) and NHP (Non-Hyperactivated, when VCL<190.17 and ALH<6.44 µm). A T test and Pearson's correlation were performed with a significance of p<0.05. Among the groups, were detected higher values of total (HP: 68,2% vs NHP: 35,9%) and progressive (HP:40,1% vs NPH: 17,18%) motility, VAP (HP: 165,85µm/s vs NHP: 97,72 µm/s), VSL (HP:137,63µm/s vs NHP 82,6µm/s), VCL (HP: 237,31µm/s vs NHP: 147,94µm/s) and ALH (HP: 7,28µm vs NHP:5,42µm) for the HP group. There was a correlation in the HP ejaculates between total and progressive motility. In the NHP group, a correlation was observed between motility and progressive motility, and between progressive motility and STR and LIN. It was concluded that HP spermatozoa have a higher curvilinear speed, while NHP spermatozoa stand out due to their straight path.

Características do comportamento hiperativado de espermatozoides de felinos domésticos / Characteristics of the cat sperm hyperactivated behavior

Sperm hyperactivity is a physiological behavior, and in the feline species it is characterized by an increase in the curvilinear velocity (VCL) and a greater head beat (ALH) evaluated by the CASA system. The study aimed to compare and correlate kinetic parameters of Hyperactive and Non-Hyperactive feline ejaculates when considering the means of VCL and ALH. Ejaculates were collected by electro ejaculation from 21 cats. The seminal samples had the kinetic parameters evaluated by the CASA system. From the average values of VCL and ALH, the ejaculates were classified in group HP (Hyperactivated, when VCL>190.17µm/s and ALH>6.44µm) and NHP (Non-Hyperactivated, when VCL<190.17 and ALH<6.44 µm). A T test and Pearson's correlation were performed with a significance of p<0.05. Among the groups, were detected higher values of total (HP: 68,2% vs NHP: 35,9%) and progressive (HP:40,1% vs NPH: 17,18%) motility, VAP (HP: 165,85µm/s vs NHP: 97,72 µm/s), VSL (HP:137,63µm/s vs NHP 82,6µm/s), VCL (HP: 237,31µm/s vs NHP: 147,94µm/s) and ALH (HP: 7,28µm vs NHP:5,42µm) for the HP group. There was a correlation in the HP ejaculates between total and progressive motility. In the NHP group, a correlation was observed between motility and progressive motility, and between progressive motility and STR and LIN. It was concluded that HP spermatozoa have a higher curvilinear speed, while NHP spermatozoa stand out due to their straight path.(AU)

Avaliação do perímetro escrotal e características seminais de touros senepol, de acordo com a idade / Evaluation of the scrotal perimeter and seminal characteristics of Senepol bulls according to age

O objetivo do estudo foi avaliar perímetro escrotal e características seminais de touros da raça Senepol, de acordo com a idade e as correlações entre elas. Foram avaliados 397 touros na raça Senepol, com idade entre 12 e 30 meses, puros de origem, do estado de Minas Gerais. O método de coleta foi massagem das ampolas e eletroejaculação. Foram avaliados motilidade e vigor do sêmen e os defeitos espermáticos. Os dados foram analisados pelo teste de Kruskal-Wallis e pelo método de Spearman. Os testículos apresentaram um crescimento linear com a idade. Houve maior diferença na motilidade espermática em animais com 18 e 20 meses de idade e o vigor espermático apresentou maior diferença em animais com 20 meses. Houve correlação significativa entre todas as características analisadas. O perímetro escrotal em touros Senepol apresentou crescimento linear dos 12 aos 30 meses e mesmo os animais mais jovens, com menor perímetro escrotal, já apresentaram boa qualidade de sêmen, o que mostra a precocidade reprodutiva dos machos dessa raça. Conclui-se que o perímetro escrotal em touros Senepol apresenta crescimento linear dos 12 aos 30 meses e mesmo os animais mais jovens, já apresentam boa qualidade de sêmen, indicando uma precocidade reprodutiva dos machos dessa raça.

The objective of this study was evaluate scrotal perimeter and seminal characteteristics of Senepol bulls acording to age and correlations between them. 397 Senepol bulls were avaliable, with ages between 12 and 30 months, pure breed, in the state of Minas Gerais. The collect method were massage of ampoules and electro-ejaculation. Motility and vigor of semen and sperm defects were avaliable. Data were analyzed by the Kruskal-Wallis test and the Spearman method. The tests had a linear growth with age. The biggest difference of motility were in animals with 18 and 20 months of age and the biggest difference of vigor were in animals with 20 months. There was a significant correlation between all analyzed characteristics. Concludes that the scrotal perimeter in Senepol bulls have a linear growth from 12 to 30 months and even the youngest animals, it is a already a good semen quality, indicating a reproductive precocity in males of this breed.

Avaliação do perímetro escrotal e características seminais de touros senepol, de acordo com a idade / Evaluation of the scrotal perimeter and seminal characteristics of Senepol bulls according to age

O objetivo do estudo foi avaliar perímetro escrotal e características seminais de touros da raça Senepol, de acordo com a idade e as correlações entre elas. Foram avaliados 397 touros na raça Senepol, com idade entre 12 e 30 meses, puros de origem, do estado de Minas Gerais. O método de coleta foi massagem das ampolas e eletroejaculação. Foram avaliados motilidade e vigor do sêmen e os defeitos espermáticos. Os dados foram analisados pelo teste de Kruskal-Wallis e pelo método de Spearman. Os testículos apresentaram um crescimento linear com a idade. Houve maior diferença na motilidade espermática em animais com 18 e 20 meses de idade e o vigor espermático apresentou maior diferença em animais com 20 meses. Houve correlação significativa entre todas as características analisadas. O perímetro escrotal em touros Senepol apresentou crescimento linear dos 12 aos 30 meses e mesmo os animais mais jovens, com menor perímetro escrotal, já apresentaram boa qualidade de sêmen, o que mostra a precocidade reprodutiva dos machos dessa raça. Conclui-se que o perímetro escrotal em touros Senepol apresenta crescimento linear dos 12 aos 30 meses e mesmo os animais mais jovens, já apresentam boa qualidade de sêmen, indicando uma precocidade reprodutiva dos machos dessa raça.(AU)

The objective of this study was evaluate scrotal perimeter and seminal characteteristics of Senepol bulls acording to age and correlations between them. 397 Senepol bulls were avaliable, with ages between 12 and 30 months, pure breed, in the state of Minas Gerais. The collect method were massage of ampoules and electro-ejaculation. Motility and vigor of semen and sperm defects were avaliable. Data were analyzed by the Kruskal-Wallis test and the Spearman method. The tests had a linear growth with age. The biggest difference of motility were in animals with 18 and 20 months of age and the biggest difference of vigor were in animals with 20 months. There was a significant correlation between all analyzed characteristics. Concludes that the scrotal perimeter in Senepol bulls have a linear growth from 12 to 30 months and even the youngest animals, it is a already a good semen quality, indicating a reproductive precocity in males of this breed.(AU)

Exame andrológico e criopreservação de sêmen em felídeos selvagens / Andrological evaluation and semen cryopreservation in wild felids

Uma primeira etapa básica e essencial para a aplicação de biotécnicas da reprodução é a coleta eavaliação do sêmen, seguida da sua criopreservação. Apresentamos aqui alguns protocolos, com base emresultados publicados e observações pessoais, para as diferentes espécies de felídeos selvagens. Para felídeosdomésticos e selvagens, a eletroejaculação é o método mais frequentemente utilizado para coleta de sêmen. Acoleta por vagina artificial, algumas vezes empregada em gatos domésticos e já descrita para guepardos, exige ocondicionamento do animal e a habituação ao coletor. A cateterização uretral, associada a fármacos quepromovem a ejaculação, surge também como uma outra opção para coleta de sêmen.(AU)

A basic and essential first step in the application of reproduction biotechniques is the semen collectionand evaluation, followed by its cryopreservation. We present here some protocols, based on published resultsand personal observations, for the different species of wild felids. For domestic and wild felids,electroejaculation is the most frequently used method for collecting semen. Collection by artificial vagina,sometimes used in domestic cats and already described for cheetahs, requires the conditioning of the animal andhabituation to the collector. Urethral catheterization, associated with drugs that promote ejaculation, alsoappears as another option for semen collection.(AU)

Exame andrológico e criopreservação de sêmen em felídeos selvagens / Andrological evaluation and semen cryopreservation in wild felids

Uma primeira etapa básica e essencial para a aplicação de biotécnicas da reprodução é a coleta eavaliação do sêmen, seguida da sua criopreservação. Apresentamos aqui alguns protocolos, com base emresultados publicados e observações pessoais, para as diferentes espécies de felídeos selvagens. Para felídeosdomésticos e selvagens, a eletroejaculação é o método mais frequentemente utilizado para coleta de sêmen. Acoleta por vagina artificial, algumas vezes empregada em gatos domésticos e já descrita para guepardos, exige ocondicionamento do animal e a habituação ao coletor. A cateterização uretral, associada a fármacos quepromovem a ejaculação, surge também como uma outra opção para coleta de sêmen.

A basic and essential first step in the application of reproduction biotechniques is the semen collectionand evaluation, followed by its cryopreservation. We present here some protocols, based on published resultsand personal observations, for the different species of wild felids. For domestic and wild felids,electroejaculation is the most frequently used method for collecting semen. Collection by artificial vagina,sometimes used in domestic cats and already described for cheetahs, requires the conditioning of the animal andhabituation to the collector. Urethral catheterization, associated with drugs that promote ejaculation, alsoappears as another option for semen collection.

EVOLUÇÃO APÓS QUATRO ANOS DE APLICAÇÃO DA CAPACIDADE ANDROLÓGICA POR PONTOS (CAP) EM TOUROS NELORE E ESTUDO DA RELAÇÃO COM A PRENHEZ EM ESTAÇÃO DE ACASALAMENTO

Abstract We studied the evolution and the relationship between the fertility of a cow herd and the breeding soundness evaluation (BSE). During four years, 1092 Nellore zebu bulls were maintained on pasture conditions in Mato Grosso do Sul State - Brazil. After the clinic evaluation, scrotal circumference was measured and semen was collected by electro ejaculation and analyzed for physical and morphological aspects. Scores for CAP were established by sperm progressive motility (MOT), sperm morphology (MORF), and scrotal circumference (PEP). Modifications to original BSE were established by sperm concentration, sperm motility intensity, and testicles morphology. During the breeding season, 157 bulls were chosen from a-c BSE score. Fifteen reproductive groups of bulls were established randomly among three groups of females (Nellore zebu heifers, first-calf cows, mature cows, and first-calf cross-bred cows). After four years of observations, there was a significant increase in BSE varying, respectively, from 59.5 to 79.0 in 1994 to 1997. The BSE scores PEP, MOT and MORF increased in the period from 17.5, 12.7, and 29.3 to 30.1, 14.8, and 34.4, respectively. Correlations were observed among the BSE components with the year and the studied reproductive characteristics. There was a significant increase of bulls in the BSE categories 'a' and 'b' of 9.0% and 46.9% vs. 43.5% and 44.8%, respectively. The data of 1997 breeding season were: 82.0 ± 13.2 points and 83.1 ± 8.4% for BSE and pregnancy rate, respectively. There was a relationship (P 0.001) between BSE and pregnancy rate (r=0.25). The Breeding Soundness Evaluations (BSE) showed a positive correlation with the pregnancy rate in a herd with different categories of females. The use of 'a' and 'b' BSE bull groups is compatible with the high fertility potential of the herd. The ratio bull:cow used underestimates the reproductive potential of the bulls' fertility. The technique used was important to evaluate the group potential fertility and to modify the reproductive characteristics of bulls herd.

Resumo Estudou-se a evolução da capacidade andrológica por pontos (CAP) de 1092 touros Nelore, mantidos a pasto, durante quatro anos, e sua relação com a porcentagem de prenhez após uma estação reprodutiva. A cada ano, realizaram-se avaliações clínico-andrológicas em que mediu-se o perímetro escrotal (PE) e se coleou sêmen por eletroejaculação, para avaliação dos aspectos físicos e morfológicos dos ejaculados. A partir dos pontos obtidos pelo PE (PEP), motilidade progressiva (MOT) e morfologia espermática (MORF), computaram-se os CAP. Na estação de monta foram utilizadas 4319 fêmeas e 256 touros Nelore, em regime de pasto, para avaliação da fertilidade do rebanho. Após a escolha de 157 touros dentro das faixas A-C do CAP, foram estabelecidos, aleatoriamente, 15 grupos de touros para diferentes categorias e quantidade de fêmeas. Nos quatro anos de estudo houve aumento significativo do CAP, de 59,5 a 79,0. A PEP, MOT e MORF aumentaram no mesmo período de 17,5, 12,7 e 29,3 para 30,1, 14,8 e 34,4, respectivamente (P 0,05). Foram observadas correlações entre o CAP com os seus componentes, o ano e as características reprodutivas estudadas. Houve aumento significativo de touros enquadrados nas categorias A e B do CAP de 9,0% e 46,9% vs. 43,5% e 44,8%, respectivamente. O CAP e a porcentagem de prenhez das fêmeas foram de 82,0 ± 13,2 pontos e 83,1 ± 8,4 %, respectivamente. Houve correlação (P 0,001) entre o CAP e a taxa de prenhez (r=0,25). A utilização de grupos de touros das faixas A e B do CAP favoreceu o aumento da percentagem de prenhez no rebanho. A relação touro:fêmea observada sub-utiliza o potencial reprodutivo dos touros. A técnica empregada foi importante para avaliar a fertilidade potencial e modificar as características reprodutivas da população de touros estudada.

Técnicas reprodutivas para a conservação de felídeos silvestres / Reproductive techniques for the conservation of wild felids

Os felídeos, principalmente os grandes, muitas vezes são vistos como ícones ou, na terminologiaacadêmica, como espécie bandeira simbolizando o ecossistema. Para que as técnicas de reprodução assistida(TRAs) possam desempenhar um papel significativo na conservação de felídeos, é essencial que as técnicas maisfundamentais de IA, FIV e TE, além da criopreservação de espermatozoides e embriões, primeiro sejamcompletamente desenvolvidas via estudos sistemáticos. Para a coleta de sêmen em felídeos, geralmente tem sidousada a eletroejaculação ou a coleta farmacológica, com fármaco que induz a ejaculação. Uma abordagem parasuperar a baixa qualidade do sêmen após a criopreservação, tem sido utilizar a inseminação laparoscópica, quepossibilita depositar o sêmen no ápice do corno uterino ou mesmo no oviduto, para alcançar uma fertilidade maissatisfatória. Considerando que os felídeos geralmente têm uma expectativa de vida curta, baixa produção defilhotes (1-4 filhotes/ninhada) e início precoce da senescência reprodutiva (~7-10 anos de idade), a atualmanutenção de populações geneticamente viáveis em cativeiro é muito difícil. As TRAs oferecem umaferramenta potencial para enfrentar esses desafios de manejo e auxiliar na conservação de felídeos silvestres.(AU)

Felids, especially the large species, are often seen as icons or, in academic terminology, as flag speciessymbolizing the ecosystem. For assisted reproductive techniques (ARTs) to play a significant role in theconservation of felids, it is essential that the most fundamental techniques of AI, IVF and ET, in addition tocryopreservation of sperm and embryos, be first fully developed through systematic studies. For the semencollection in felids, electroejaculation or pharmacological collection has usually been used, with a drug thatinduces ejaculation. One approach to overcome the poor quality of semen after cryopreservation has been to uselaparoscopic insemination, which makes it possible to deposit the semen at the apex of the uterine horn or evenin the oviduct, to achieve a more satisfactory fertility. Considering that felids generally have a short lifeexpectancy, low pups production (1-4 pups/litter) and early onset of reproductive senescence (~ 7-10 years ofage), the current maintenance of genetically viable populations in captivity is very difficult. ARTs offer apotential tool to address these management challenges and assist in the conservation of wild felids.(AU)

Cooling of canine semen contaminated with urine / Resfriamento do sêmen canino contaminado por urina

Contamination of canine semen with urine, caused by urination during ejaculation or semen flow into the urinary bladder, is a common problem of sperm collected by electroejaculation for cryopreservation. The effects of urine on sperm cryopreservation, however, are not fully understood. The present study determined the acceptable upper level of contamination with urine for canine semen preservation after 24-h cooling. Semen samples obtained from 10 dogs were incubated with Ringers solution and different urine concentrations (0, 3, 6, 13, 25 50% and 100%) at 37C for 20 min. The samples were centrifuged and the decanted sperm resuspended and cooled to 5C for up to 24 h in a milk-based semen extender. The quality of the contaminated semen samples only differed from that of the control treatment after exposure to urine concentrations above 13%. After cooling, only sperm incubated with urine solutions above 25% exhibited lower activity than those incubated in simple Ringers solution. In conclusion, canine semen resists contamination with up to 13% urine for 20 min, and the motility of samples contaminated with up to 25% recovers similarly to that of uncontaminated samples if cooled at 5C for 24 h in a milk-based extender.(AU)

A contaminação do sêmen canino pode ser causada pela micção durante a ejaculação, ou quando o sêmen flui diretamentepara a bexiga, que são problemas comuns durante a eletroejaculação para a criopreservação. Os efeitos que a urina causaao processo de criopreservação, entretanto, não estão completamente entendidos. O presente estudo determinou a resistênciado sêmen a diferentes níveis de contaminação por urina após 24h de resfriamento. As amostras de sêmen obtidas de 10 cãesforam incubadas em soluções de Ringer, e em diferentes concentrações de urina (0,3,6,13,25,50 e 100%), a 37°C, por 20 min. Asamostras foram centrifugadas e os espermatozoides decantados foram ressuspendidos, e resfriados a 5°C por 24h em diluidor abase de leite. A qualidade das amostras de sêmen contaminado por urina diferiu do controle apenas após a exposição de soluçõescom concentração superior a 13% de urina. Após o resfriamento, apenas os espermatozoides incubados com soluções de urinasuperiores a 25% exibiram menor atividade que as incubadas na solução simples de Ringer. Em conclusão, o sêmen canino resisteà contaminação com até 13% de urina durante 20 minutos e a motilidade das amostras contaminadas com até 25% recuperam-sede forma semelhante às amostras não contaminadas, se resfriadas a 5°C durante 24 h em um diluidor à base de leite.(AU)

Técnicas reprodutivas para a conservação de felídeos silvestres / Reproductive techniques for the conservation of wild felids

Os felídeos, principalmente os grandes, muitas vezes são vistos como ícones ou, na terminologiaacadêmica, como espécie bandeira simbolizando o ecossistema. Para que as técnicas de reprodução assistida(TRAs) possam desempenhar um papel significativo na conservação de felídeos, é essencial que as técnicas maisfundamentais de IA, FIV e TE, além da criopreservação de espermatozoides e embriões, primeiro sejamcompletamente desenvolvidas via estudos sistemáticos. Para a coleta de sêmen em felídeos, geralmente tem sidousada a eletroejaculação ou a coleta farmacológica, com fármaco que induz a ejaculação. Uma abordagem parasuperar a baixa qualidade do sêmen após a criopreservação, tem sido utilizar a inseminação laparoscópica, quepossibilita depositar o sêmen no ápice do corno uterino ou mesmo no oviduto, para alcançar uma fertilidade maissatisfatória. Considerando que os felídeos geralmente têm uma expectativa de vida curta, baixa produção defilhotes (1-4 filhotes/ninhada) e início precoce da senescência reprodutiva (~7-10 anos de idade), a atualmanutenção de populações geneticamente viáveis em cativeiro é muito difícil. As TRAs oferecem umaferramenta potencial para enfrentar esses desafios de manejo e auxiliar na conservação de felídeos silvestres.

Felids, especially the large species, are often seen as icons or, in academic terminology, as flag speciessymbolizing the ecosystem. For assisted reproductive techniques (ARTs) to play a significant role in theconservation of felids, it is essential that the most fundamental techniques of AI, IVF and ET, in addition tocryopreservation of sperm and embryos, be first fully developed through systematic studies. For the semencollection in felids, electroejaculation or pharmacological collection has usually been used, with a drug thatinduces ejaculation. One approach to overcome the poor quality of semen after cryopreservation has been to uselaparoscopic insemination, which makes it possible to deposit the semen at the apex of the uterine horn or evenin the oviduct, to achieve a more satisfactory fertility. Considering that felids generally have a short lifeexpectancy, low pups production (1-4 pups/litter) and early onset of reproductive senescence (~ 7-10 years ofage), the current maintenance of genetically viable populations in captivity is very difficult. ARTs offer apotential tool to address these management challenges and assist in the conservation of wild felids.

Evolution after four years of breeding soundness evaluation (BSE) in Nellore zebu bulls and assessment of the relationship with fertility in a breeding season / Evolução após quatro anos de aplicação da capacidade andrológica por pontos (CAP) em touros Nelore e estudo da relação com a prenhez em estação de acasalamento

We studied the evolution and the relationship between the fertility of a cow herd and the breeding soundness evaluation (BSE). During four years, 1092 Nellore zebu bulls were maintained on pasture conditions in Mato Grosso do Sul State – Brazil. After the clinic evaluation, scrotal circumference was measured and semen was collected by electro ejaculation and analyzed for physical and morphological aspects. Scores for CAP were established by sperm progressive motility (MOT), sperm morphology (MORF), and scrotal circumference (PEP). Modifications to original BSE were established by sperm concentration, sperm motility intensity, and testicles morphology. During the breeding season, 157 bulls were chosen from a-c BSE score. Fifteen reproductive groups of bulls were established randomly among three groups of females (Nellore zebu heifers, first-calf cows, mature cows, and first-calf cross-bred cows). After four years of observations, there was a significant increase in BSE varying, respectively, from 59.5 to 79.0 in 1994 to 1997. The BSE scores PEP, MOT and MORF increased in the period from 17.5, 12.7, and 29.3 to 30.1, 14.8, and 34.4, respectively. Correlations were observed among the BSE components with the year and the studied reproductive characteristics. There was a significant increase of bulls in the BSE categories 'a' and 'b' of 9.0% and 46.9% vs. 43.5% and 44.8%, respectively. The data of 1997 breeding season were: 82.0 ± 13.2 points and 83.1 ± 8.4% for BSE and pregnancy rate, respectively. There was a relationship (P <0.001) between BSE and pregnancy rate (r=0.25). The Breeding Soundness Evaluations (BSE) showed a positive correlation with the pregnancy rate in a herd with different categories of females. The use of 'a' and 'b' BSE bull groups is compatible with the high fertility potential of the herd. The ratio bull:cow used underestimates the reproductive potential of the bulls’ fertility. [...]

Estudou-se a evolução da capacidade andrológica por pontos (CAP) de 1092 touros Nelore, mantidos a pasto, durante quatro anos, e sua relação com a porcentagem de prenhez após uma estação reprodutiva. A cada ano, realizaram-se avaliações clínico-andrológicas em que mediu-se o perímetro escrotal (PE) e se coleou sêmen por eletroejaculação, para avaliação dos aspectos físicos e morfológicos dos ejaculados. A partir dos pontos obtidos pelo PE (PEP), motilidade progressiva (MOT) e morfologia espermática (MORF), computaram-se os CAP. Na estação de monta foram utilizadas 4319 fêmeas e 256 touros Nelore, em regime de pasto, para avaliação da fertilidade do rebanho. Após a escolha de 157 touros dentro das faixas A-C do CAP, foram estabelecidos, aleatoriamente, 15 grupos de touros para diferentes categorias e quantidade de fêmeas. Nos quatro anos de estudo houve aumento significativo do CAP, de 59,5 a 79,0. A PEP, MOT e MORF aumentaram no mesmo período de 17,5, 12,7 e 29,3 para 30,1, 14,8 e 34,4, respectivamente (P<0,05). Foram observadas correlações entre o CAP com os seus componentes, o ano e as características reprodutivas estudadas. Houve aumento significativo de touros enquadrados nas categorias A e B do CAP de 9,0% e 46,9% vs. 43,5% e 44,8%, respectivamente. O CAP e a porcentagem de prenhez das fêmeas foram de 82,0 ± 13,2 pontos e 83,1 ± 8,4 %, respectivamente. Houve correlação (P<0,001) entre o CAP e a taxa de prenhez (r=0,25). A utilização de grupos de touros das faixas A e B do CAP favoreceu o aumento da percentagem de prenhez no rebanho. A relação touro:fêmea observada sub-utiliza o potencial reprodutivo dos touros. A técnica empregada foi importante para avaliar a fertilidade potencial e modificar as características reprodutivas da população de touros estudada.

Reproductive characteristics of Crioulo Lageano breed bulls (Bos taurus) at puberty

The aim of this research was to evaluate pubertal development characteristics of Crioulo Lageano breed bulls (n = 10) using morphometric measurements and semen analysis, to identify factors that can be used to estimate age at puberty in this locally adapted breed. Monthly measurement of body weight and fortnightly measurement of scrotal circumference, chest girth, testicular length, width, thickness, and volume were recorded for each of the 10 Crioulo Lageano breed bulls, which were between 10 and 20 months old. During this period, semen samples were collected every two weeks using electro ejaculation method and analyzed physically and morphologically. The ages of the appearance of first spermatozoa in the ejaculate (FSE), the first motile spermatozoa in the ejaculate (FSEM), seminal puberty (PUB) and total detachment between glans penis and prepuce (DPP) were ascertained. Crioulo Lageano bulls reached puberty at 14.1 ± 2.0 months old with lower weight and larger testicles than those of other bovine breeds. Similarly, the period from FSEM to PUB was shorter and the period from FSE to FSEM was longer than those reported for other breeds. The most important characteristics studied in order to estimate puberty age in Crioulo Lageano bulls were the measures of testicular length, width, and volume. These parameters can be used as criteria to select young bulls as sires.

Reproductive characteristics of Crioulo Lageano breed bulls (Bos taurus) at puberty

The aim of this research was to evaluate pubertal development characteristics of Crioulo Lageano breed bulls (n = 10) using morphometric measurements and semen analysis, to identify factors that can be used to estimate age at puberty in this locally adapted breed. Monthly measurement of body weight and fortnightly measurement of scrotal circumference, chest girth, testicular length, width, thickness, and volume were recorded for each of the 10 Crioulo Lageano breed bulls, which were between 10 and 20 months old. During this period, semen samples were collected every two weeks using electro ejaculation method and analyzed physically and morphologically. The ages of the appearance of first spermatozoa in the ejaculate (FSE), the first motile spermatozoa in the ejaculate (FSEM), seminal puberty (PUB) and total detachment between glans penis and prepuce (DPP) were ascertained. Crioulo Lageano bulls reached puberty at 14.1 ± 2.0 months old with lower weight and larger testicles than those of other bovine breeds. Similarly, the period from FSEM to PUB was shorter and the period from FSE to FSEM was longer than those reported for other breeds. The most important characteristics studied in order to estimate puberty age in Crioulo Lageano bulls were the measures of testicular length, width, and volume. These parameters can be used as criteria to select young bulls as sires.(AU)

Evolution after four years of breeding soundness evaluation (BSE) in Nellore zebu bulls and assessment of the relationship with fertility in a breeding season / Evolução após quatro anos de aplicação da capacidade andrológica por pontos (CAP) em touros Nelore e estudo da relação com a prenhez em estação de acasalamento

We studied the evolution and the relationship between the fertility of a cow herd and the breeding soundness evaluation (BSE). During four years, 1092 Nellore zebu bulls were maintained on pasture conditions in Mato Grosso do Sul State Brazil. After the clinic evaluation, scrotal circumference was measured and semen was collected by electro ejaculation and analyzed for physical and morphological aspects. Scores for CAP were established by sperm progressive motility (MOT), sperm morphology (MORF), and scrotal circumference (PEP). Modifications to original BSE were established by sperm concentration, sperm motility intensity, and testicles morphology. During the breeding season, 157 bulls were chosen from a-c BSE score. Fifteen reproductive groups of bulls were established randomly among three groups of females (Nellore zebu heifers, first-calf cows, mature cows, and first-calf cross-bred cows). After four years of observations, there was a significant increase in BSE varying, respectively, from 59.5 to 79.0 in 1994 to 1997. The BSE scores PEP, MOT and MORF increased in the period from 17.5, 12.7, and 29.3 to 30.1, 14.8, and 34.4, respectively. Correlations were observed among the BSE components with the year and the studied reproductive characteristics. There was a significant increase of bulls in the BSE categories 'a' and 'b' of 9.0% and 46.9% vs. 43.5% and 44.8%, respectively. The data of 1997 breeding season were: 82.0 ± 13.2 points and 83.1 ± 8.4% for BSE and pregnancy rate, respectively. There was a relationship (P <0.001) between BSE and pregnancy rate (r=0.25). The Breeding Soundness Evaluations (BSE) showed a positive correlation with the pregnancy rate in a herd with different categories of females. The use of 'a' and 'b' BSE bull groups is compatible with the high fertility potential of the herd. The ratio bull:cow used underestimates the reproductive potential of the bulls fertility. [...](AU)

Estudou-se a evolução da capacidade andrológica por pontos (CAP) de 1092 touros Nelore, mantidos a pasto, durante quatro anos, e sua relação com a porcentagem de prenhez após uma estação reprodutiva. A cada ano, realizaram-se avaliações clínico-andrológicas em que mediu-se o perímetro escrotal (PE) e se coleou sêmen por eletroejaculação, para avaliação dos aspectos físicos e morfológicos dos ejaculados. A partir dos pontos obtidos pelo PE (PEP), motilidade progressiva (MOT) e morfologia espermática (MORF), computaram-se os CAP. Na estação de monta foram utilizadas 4319 fêmeas e 256 touros Nelore, em regime de pasto, para avaliação da fertilidade do rebanho. Após a escolha de 157 touros dentro das faixas A-C do CAP, foram estabelecidos, aleatoriamente, 15 grupos de touros para diferentes categorias e quantidade de fêmeas. Nos quatro anos de estudo houve aumento significativo do CAP, de 59,5 a 79,0. A PEP, MOT e MORF aumentaram no mesmo período de 17,5, 12,7 e 29,3 para 30,1, 14,8 e 34,4, respectivamente (P<0,05). Foram observadas correlações entre o CAP com os seus componentes, o ano e as características reprodutivas estudadas. Houve aumento significativo de touros enquadrados nas categorias A e B do CAP de 9,0% e 46,9% vs. 43,5% e 44,8%, respectivamente. O CAP e a porcentagem de prenhez das fêmeas foram de 82,0 ± 13,2 pontos e 83,1 ± 8,4 %, respectivamente. Houve correlação (P<0,001) entre o CAP e a taxa de prenhez (r=0,25). A utilização de grupos de touros das faixas A e B do CAP favoreceu o aumento da percentagem de prenhez no rebanho. A relação touro:fêmea observada sub-utiliza o potencial reprodutivo dos touros. A técnica empregada foi importante para avaliar a fertilidade potencial e modificar as características reprodutivas da população de touros estudada.(AU)

Avaliação andrológica de ovinos do Núcleo de Ruminantes da UEMA Campus Paulo VI / Evaluation andrologic of Ruminant Center of cattle from UEMA - Campus Paulo VI

The aim of the study was to characterize andrologicaly sheeps belonging to Ruminants Center ofUniversidade Estadual do Maranhao. The semen collection was performed by electro ejaculation. The bodycondition score from seven animals showed averaging 4.57 ± 1.13. The scrotal circumference (SC) was 30.58 ±4.91 cm, a scrotal circumference of rams presented mean length and width of the right testicle of 9.43 ± 1.16 cmand 5.44 ± 0.46 cm and the left testicle of 9.0 ± 0.81 cm and 5.51 ± 0.41 cm, respectively. The average volume ofsemen obtained was 0.89 ± 0.32 mL, appearance, color and odor sui generis were for the species. For physicalsemen parameters, the average turbulence was 3.57 ± 0.53; force of 3.14 ± 0.38; Sperm motility of 81.43 ±6.90%; sperm concentration of 0.8 ± 0.18 x109sptz / mL. The proportion of living and dead sperm stained witheosin-nigrosin averaged 74.1 ± 3.02% and 25 ± 3.46%, respectively. According to this study and the resultsobtained, sheep showed morphology and physical aspects consistent with the species and can be used asbreeders.(AU)

Avaliação andrológica de ovinos do Núcleo de Ruminantes da UEMA – Campus Paulo VI / Evaluation andrologic of Ruminant Center of cattle from UEMA - Campus Paulo VI

The aim of the study was to characterize andrologicaly sheeps belonging to Ruminants Center ofUniversidade Estadual do Maranhao. The semen collection was performed by electro ejaculation. The bodycondition score from seven animals showed averaging 4.57 ± 1.13. The scrotal circumference (SC) was 30.58 ±4.91 cm, a scrotal circumference of rams presented mean length and width of the right testicle of 9.43 ± 1.16 cmand 5.44 ± 0.46 cm and the left testicle of 9.0 ± 0.81 cm and 5.51 ± 0.41 cm, respectively. The average volume ofsemen obtained was 0.89 ± 0.32 mL, appearance, color and odor sui generis were for the species. For physicalsemen parameters, the average turbulence was 3.57 ± 0.53; force of 3.14 ± 0.38; Sperm motility of 81.43 ±6.90%; sperm concentration of 0.8 ± 0.18 x109sptz / mL. The proportion of living and dead sperm stained witheosin-nigrosin averaged 74.1 ± 3.02% and 25 ± 3.46%, respectively. According to this study and the resultsobtained, sheep showed morphology and physical aspects consistent with the species and can be used asbreeders.

Establishment of an anesthetic protocol for semen collection by electroejaculation in six-banded armadillos (Euphractus sexcinctus Linnaeus, 1758) / Estabelecimento de um protocolo anestésico para coleta de sêmen por eletroejaculação em tatus-peba (Euphractus sexcinctus Linnaeus, 1758)

The aim was to verify the effects of different anesthetic protocols used during electroejaculation (EEJ) in six-banded armadillos (Euphractus sexcinctus). Four sexually matured animals were physically restrained and subjected to semen collection by the EEJ following three treatments: The control group consisted of no use of anesthesia; in the others, the anesthetic combinations xylazine/ketamine/propofol or butorphanol/ ketamine/propofol were administered. For each group, twelve procedures were conducted for EEJ. Semen was evaluated for volume, color, aspect, motility, sperm concentration, morphology, viability, and functional membrane integrity. The highest efficiency (100% ejaculates) was achieved when the control group was used; the xylazine/ketamine/propofol association provided only 11 ejaculates from a total of 12 attempts (91.6% efficiency), while only 4 ejaculates (33% efficiency) were obtained with butorphanol/ketamine/propofol (P<0.05). Both protocols provided rapid induction and relaxation enough to perform the EEJ. In the use of butorphanol/ketamine/propofol, the animals recovered at 16.5±1.5min, a time shorter than in the use of xylazine/ketamine/propofol protocol, 20.7±1.0min (P>0.05). The semen volume and sperm concentration obtained in the use of xylazine/ketamine/propofol association were significantly higher than those verified for butorphanol/ketamine/propofol protocol. In conclusion, the xylazine/ketamine/propofol association is indicated for anesthesia of six-banded armadillos submitted to EEJ.(AU)

Objetivou-se verificar os efeitos de diferentes protocolos anestésicos usados durante a eletroejaculação (EEJ) em tatus-peba (Euphractus sexcinctus). Quatro animais sexualmente maduros foram contidos fisicamente e submetidos à coleta de sêmen por EEJ, seguindo três tratamentos: o grupo controle consistiu do não uso de anestesia; nos outros, foram administradas combinações anestésicas de xilazina/cetamina/propofol, ou butorfanol/cetamina/propofol. Para cada grupo, foram conduzidos 12 procedimentos de EEJ. O sêmen foi avaliado para volume, cor, aspecto, motilidade, concentração de espermatozoides, morfologia, viabilidade e integridade funcional da membrana. A mais alta eficiência (100% de ejaculados) foi alcançada quando o grupo controle foi utilizado; a associação de cetamina/xilazina/propofol forneceu apenas 11 ejaculados de um total de 12 tentativas (de eficiência 91,6%), enquanto apenas quatro ejaculados (eficiência de 33%) foram obtidos com butorfanol/cetamina/propofol (P<0,05). Ambos os protocolos forneceram rápida indução e relaxamento suficientes para executar a EEJ. Na utilização de butorfanol/cetamina/propofol, os animais se recuperaram em 16,5±1,5min, um tempo mais curto do que no uso de xilazina/cetamina/protocolo de propofol, 20,7±1,0min (P>0,05). O volume de sêmen e a concentração espermática obtidos no uso da associação xilazina/cetamina/propofol foram significativamente maiores do que os verificados para o protocolo butorfanol/cetamina/propofol. Em conclusão, a associação de cetamina/xilazina/propofol é indicada para anestesia de tatus-peba submetidos à EEJ.(AU)