Use of alternative diluent ACP-Lact® and coconut water powder (ACP) in cryopreservation of goat semen and fixed-time artificial insemination (FTAI) in half-breed goats.

The aim of the present study was to evaluate semen cryopreservation with ACP-Lact® diluent, which consists of coconut water powder (ACP) added to goat milk powder. After thawing, the samples were evaluated for sperm kinetics, membrane evaluation and in vivo insemination. For cryopreservation, a pool was made with the ejaculate of six goats, diluted in four equal aliquots for the respective treatments: T1 (ACP-Lact®); T2 (ACP-Lact® 50%); T3 (ACP + 2.5% egg yolk) and T4 (Tris + 2.5% egg yolk). After dilution of the treatments, the samples were placed in 0.5 ml straws and chilled at a rate of -1.07°C/min. After reaching 4°C and stabilizing for one hour, the straws were placed in nitrogen vapour at -60°C for 15 minutes and then immersed in liquid nitrogen (-196ºC). The straws were thawed in a 37°C water bath and kinetic assessments were performed immediately using a computerized semen analysis program (CSA), viability (EN), membrane functionality (HOST), mitochondrial activity (DAB) and DNA integrity assessment of spermatozoa. For the in vivo experiment, ten goats were inseminated, divided into two groups of five goats each, G1 inseminated with ACP-Lact® and G2 with ACP, by fixed-time artificial insemination (FTAI). Regarding the kinetic parameters, the ACP-Lact® treatment showed higher progressive motility (PM) and sperm velocity than the other treatments (36.77%). In the VSL parameter the ACP-Lact diluent was superior to ACP and Tris. In viability the treatment with ACP-Lact® was superior to the treatment with Tris, 95% and 83% respectively. In FTAI two goats were born out of the 5 goats inseminated with ACP-Lact®. It was concluded that the use of ACP-Lact® for cryopreservation of caprine semen is efficient in maintaining seminal parameters during thawing in vitro and in vivo and proved to be a good alternative extender for the caprine species.

Assessment of coconut water added to Numida meleagris egg yolk as cryoprotectant for goat semen.

Effects were assessed of the dilutants TRIS and ACP - 101c® with the addition of different guinea fowl (Numida meleagris) egg yolk concentrations. Fifteen ejaculates were collected from five goats of the Anglo Nubian breed. The ejaculates were pooled and then divided into 12 groups, two control groups (GC1 TRIS, with 2.5% Gallus gallus domesticus hen egg yolk GOGD), (GC2 Control Group ACP - 101c®, with the addition of 2.5% Gallus gallus domesticus hen egg yolk GOGD) and ten experimental groups (EG), containing TRIS and ACP added with different concentrations of egg yolk from guinea hen (Numida meleagris) (TRIS 2,5% GONM; TRIS 5% GONM; TRIS 10% GONM; TRIS 15% GONM; TRIS 20% GONM; ACP® 2,5% GONM; ACP® 5% GONM; ACP® 10% GONM; ACP® 15% GONM; ACP® 20% GONM). Then cryopreservation was carried out and the samples stored in liquid nitrogen (-196 °C). After seven days, the samples were thawed and assessed for spermatic kinetics, immunofluorescence and sperm morphology. Analysis of GOMN by the CASA system showed that the various parameters were similar to those of GOGD (P>0.05). The membrane integrity, mitochondrial potential and the acrosome were not influenced by the treatment (P>0.05) nor by the dilutant used for cryopreservation (P>0.05). The spermatic morphology was also preserved by the different GOGD and GONM concentrations in the ACP® and TRIS dilutants, with no statistically significant differences (P<0.05). It was concluded that Numida meleagris egg yolk, as external membrane cryoproctant added to the dilutants ACP-101c® and TRIS, improved goat semen quality.

Assessment of coconut water added to Numida meleagris egg yolk as cryoprotectant for goat semen

Effects were assessed of the dilutants TRIS and ACP - 101c® with the addition of different guinea fowl (Numida meleagris) egg yolk concentrations. Fifteen ejaculates were collected from five goats of the Anglo Nubian breed. The ejaculates were pooled and then divided into 12 groups, two control groups (GC1 TRIS, with 2.5% Gallus gallus domesticus hen egg yolk GOGD), (GC2 Control Group ACP - 101c®, with the addition of 2.5% Gallus gallus domesticus hen egg yolk GOGD) and ten experimental groups (EG), containing TRIS and ACP added with different concentrations of egg yolk from guinea hen (Numida meleagris) (TRIS 2,5% GONM; TRIS 5% GONM; TRIS 10% GONM; TRIS 15% GONM; TRIS 20% GONM; ACP® 2,5% GONM; ACP® 5% GONM; ACP® 10% GONM; ACP® 15% GONM; ACP® 20% GONM). Then cryopreservation was carried out and the samples stored in liquid nitrogen (-196 °C). After seven days, the samples were thawed and assessed for spermatic kinetics, immunofluorescence and sperm morphology. Analysis of GOMN by the CASA system showed that the various parameters were similar to those of GOGD (P>0.05). The membrane integrity, mitochondrial potential and the acrosome were not influenced by the treatment (P>0.05) nor by the dilutant used for cryopreservation (P>0.05). The spermatic morphology was also preserved by the different GOGD and GONM concentrations in the ACP® and TRIS dilutants, with no statistically significant differences (P<0.05). It was concluded that Numida meleagris egg yolk, as external membrane cryoproctant added to the dilutants ACP-101c® and TRIS, improved goat semen quality.(AU)

Risk factors associated with seroprevalence of Chlamydia abortus in sheep farms in Ceará, Brazil

Background: Chlamydia abortus infections (Chlamydiosis) can cause reproductive problems in sheep, such as abortionsand birth defects, leading to farm productivity loss. The symptoms, which are similar to other reproductive diseases, andthe microbial pathogenesis make the clinical diagnosis difficult. Chlamydia abortus is a zoonotic pathogen, making it apublic health issue because it can infect and induce abortions in humans. This study investigated anti-C. abortus antibodylevels and infection risk factors in sheep in the State of Ceará, Brazil.Materials, Methods & Results: Forty-three properties from 10 municipalities in 4 mesoregions in the State of Ceará, Brazil(Sertões, metropolitan Fortaleza, North Ceará and Northwest Ceará) with sheep, goats, cattle, and horses were visited.Five hundred and four serological samples from sheep were collected and tested for anti-C. abortus antibodies using anEnzyme-linked Immunosorbent Assay (ELISA) [IDEXX®, Australia] and all procedures were performed in accordancewith the manufacturers instructions at the Clinical Pathology Laboratory of EMBRAPA Goats and Sheep (Sobral, Brazil). Individual questionnaires were completed about sheep breeding practices and to identify possible C. abortus risks.Seropositive results were found in 18.45 % (93/504 individuals) of sheep, and 88.37 % (38/43 properties) of the herds hadat least one seropositive animal. The number of seropositive individuals was significantly different between adults andewes [P < 0.01; Odds Ratio (OR) = 0.510; 95% confidence interval (CI) = 0.306 - 0.850]. Logistic regression modelingidentified a missing health certificate request for newly acquired animals as a chlamydiosis risk factor [P = 0.038; OR =2.672; 95% CI = 1.058 - 6.749].Discussion: The prevalence of anti-C. abortus...(AU)

Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

Risk factors associated with seroprevalence of Chlamydia abortus in sheep farms in Ceará, Brazil

Background: Chlamydia abortus infections (Chlamydiosis) can cause reproductive problems in sheep, such as abortionsand birth defects, leading to farm productivity loss. The symptoms, which are similar to other reproductive diseases, andthe microbial pathogenesis make the clinical diagnosis difficult. Chlamydia abortus is a zoonotic pathogen, making it apublic health issue because it can infect and induce abortions in humans. This study investigated anti-C. abortus antibodylevels and infection risk factors in sheep in the State of Ceará, Brazil.Materials, Methods & Results: Forty-three properties from 10 municipalities in 4 mesoregions in the State of Ceará, Brazil(Sertões, metropolitan Fortaleza, North Ceará and Northwest Ceará) with sheep, goats, cattle, and horses were visited.Five hundred and four serological samples from sheep were collected and tested for anti-C. abortus antibodies using anEnzyme-linked Immunosorbent Assay (ELISA) [IDEXX®, Australia] and all procedures were performed in accordancewith the manufacturer’s instructions at the Clinical Pathology Laboratory of EMBRAPA Goats and Sheep (Sobral, Brazil). Individual questionnaires were completed about sheep breeding practices and to identify possible C. abortus risks.Seropositive results were found in 18.45 % (93/504 individuals) of sheep, and 88.37 % (38/43 properties) of the herds hadat least one seropositive animal. The number of seropositive individuals was significantly different between adults andewes [P < 0.01; Odds Ratio (OR) = 0.510; 95% confidence interval (CI) = 0.306 - 0.850]. Logistic regression modelingidentified a missing health certificate request for newly acquired animals as a chlamydiosis risk factor [P = 0.038; OR =2.672; 95% CI = 1.058 - 6.749].Discussion: The prevalence of anti-C. abortus...

Serological evidence of Toxoplasma gondii occurrence in naturally infected sheep in the Teresina microregion, Piauí, Brazil

ABSTRACT Toxoplasmosis is a zoonosis of reproductive nature that is transmitted to humans mainly by ingestion of undercooked meat or drinking oocyte-contaminated water. The slaughter of small ruminants in clandestine slaughterhouses and the presence of companion animals in these locations may be epidemiologically important for the dissemination of the disease. The objective of the present study was to determine toxoplasmosis prevalence by researching anti- Toxoplasma gondii antibodies in meat sheep herds from farms in Teresina microregion, Piauí, Brazil. A total of 450 blood samples were collected from sheep of both sexes, belonging to 28 herds from the 14 municipalities that comprise the microregion. The samples were analyzed by the enzyme-linked immunosorbent assay (ELISA), and the presence of anti-T. gondii antibodies was observed in 62% (279/450) of the animals in all the municipalities and on at least one farm in each municipality. The high occurrence of positive animals is an indication of reproductive problems in these herds, characterizing a problem for both sheep rearing and public health.

Risk factors and detection of brucella ovis in naturally infected rams and ewes / Fatores de risco e detecção de brucella ovis em carneiros e ovelhas naturalmente infectados

The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.(AU)

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.(AU)

Effect of adding anethole to the cryopreservation medium (powdered coconut water) on morphology, kinetics, and oxidative stress of buck sperm / Efeito da adição de anethole ao meio de criopreserção (água de coco em pó) na morfologia, cinética e estresse oxidativo do sêmen de bode

The quality of post-thawing goat sperm is critical to the success of artificial insemination protocols and may be influenced by extenders, cryoprotectants, and antioxidant substances. Therefore, the objective of this study was to evaluate the effects of the antioxidant anethole on goat sperm diluted in preservation medium based on powdered coconut water (ACP-101c) and frozen. For that, each ejaculate was submitted to the following treatments: ACP-101c (control); control plus supplementation with 30, 300, or 2000 µg/ mL anethole. The samples were thawed and evaluated for morphology, kinetics, membrane integrity, and reactive oxygen species (ROS). The addition of anethole increased morphological abnormalities (P < 0.05), however, it did not affect sperm kinetics. Flow cytometry analysis showed that sperm cells cryopreserved with 300 µg/mL anethole had lower acrosome integrity than those cryopreserved in other treatments. Evaluation of oxidative stress revealed that cells stored in the presence of 2000 µg/mL anethole had small amounts of ROS when compared to those preserved in the control medium alone or supplemented with 300 µg/mL anethole (P < 0.05). After cryopreservation of sperm with 2000 µg/mL anethole, the highest percentage of viable sperm without ROS was observed (P < 0.05). In conclusion, despite reducing ROS levels, the supplementation of anethole in ACP-101c did not affect sperm kinetics or membrane integrity post-thawing, however, it did cause morphological damage to sperm.(AU)

A qualidade do espermatozoide caprino pós-descongelação é crítica para o sucesso dos protocolos de inseminação artificial e pode ser influenciada por extensores, crioprotetores e substâncias antioxidantes. Portanto, o objetivo deste estudo foi avaliar os efeitos do antioxidante anetole sobre espermatozoides caprinos diluídos em meio de conservação à base de água de coco em pó (ACP-101c) e congelados. Para tanto, cada ejaculado foi submetido aos seguintes tratamentos: ACP-101c (controle); controle mais suplementação com 30, 300 ou 2000 µg / mL de anetole. As amostras foram descongeladas e avaliadas quanto à morfologia, cinética, integridade de membranas e espécies reativas de oxigênio. A adição de anetole aumentou as anormalidades morfológicas (P < 0,05), no entanto, não afetou a cinética dos espermatozoides. A análise da citometria de fluxo mostrou que as células de esperma criopreservadas com 300 µg / mL anethole tinham integridade acrosma menor do que aquelas criopreservadas em outros tratamentos. A avaliação do estresse oxidativo revelou que as células armazenadas na presença de 2000 µg / mL anethole apresentaram pequenas quantidades de ROS quando comparadas às preservadas em meio de controle isoladamente ou suplementadas com 300 µg / mL anethole (P < 0,05). Após a criopreservação de espermatozoides com 2000 µg / mL anethole, observou-se a maior porcentagem de espermatozoides viáveis sem ROS (P < 0,05). A população com espermatozoides viáveis sem ROS foi maior quando utilizado 2.000 µg / mL (P < 0,05). Em conclusão, apesar de reduzir os níveis de ROS, a suplementação de anetole em ACP-101c não afetou a cinética espermática e a integridade da membrana pós-descongelação, entretanto, causou danos morfológicos nos espermatozoides.(AU)

Serological evidence of Toxoplasma gondii occurrence in naturally infected sheep in the Teresina microregion, Piauí, Brazil

Toxoplasmosis is a zoonosis of reproductive nature that is transmitted to humans mainly by ingestion of undercooked meat or drinking oocyte-contaminated water. The slaughter of small ruminants in clandestine slaughterhouses and the presence of companion animals in these locations may be epidemiologically important for the dissemination of the disease. The objective of the present study was to determine toxoplasmosis prevalence by researching anti- Toxoplasma gondii antibodies in meat sheep herds from farms in Teresina microregion, Piauí, Brazil. A total of 450 blood samples were collected from sheep of both sexes, belonging to 28 herds from the 14 municipalities that comprise the microregion. The samples were analyzed by the enzyme-linked immunosorbent assay (ELISA), and the presence of anti-T. gondii antibodies was observed in 62% (279/450) of the animals in all the municipalities and on at least one farm in each municipality. The high occurrence of positive animals is an indication of reproductive problems in these herds, characterizing a problem for both sheep rearing and public health.

Epidemiological characterization and risk factors associated with Brucella ovis infection in sheep from the states of Rio Grande do Norte, Paraíba, and Sergipe / Caracterização epidemiológica e fatores de risco associados à infecção por Brucella ovis em ovinos dos estados do Rio Grande do Norte, Paraíba e Sergipe

This study aimed to determine the prevalence of ovine brucellosis in the states of Rio Grande do Norte, Paraíba, and Sergipe. A seroepidemiological survey was carried out in 121 properties under 23 municipalities between 2011 and 2012. The 1,034 serological samples collected from the sheep were subjected to the agar gel immunodiffusion (AGID) test using kits produced by the Paraná Institute of Technology (TECPAR). Anti-Brucella ovis antibodies were observed in 5.37% (70/1,304) of the animals evaluated, and the difference in the incidence of brucellosis between the adults and young animals of both sexes, and between the males and females was statistically significant (p < 0.05). The prevalence of brucellosis in Rio Grande do Norte, Paraíba, and was Sergipe 7.66% (36/470), 5.40% (13/241) and 3.54% (21/593), respectively. Of the municipalities visited, 91.30% (21/23) harbored herds with positive serology, and 42.15% (51/121) of the properties had at least one seropositive animal. The lack of training among the workers (p ? 0.05) (odds ratio = 3.68; 95% CI = 1.08-12.60; p = 0.038) was identified as a risk factor. The results obtained in this study indicate that the sheep from Rio Grande do Norte, Paraíba, and Sergipe are infected by B. ovis, thus necessitating the adoption of proper sanitary measures for preventing the spread of the disease in the herds.(AU)

O estudo teve como objetivo determinar a prevalência da Brucelose Ovina nos Estados do Rio Grandedo Norte, Paraíba e Sergipe. Foi realizado o levantamento soroepidemiológico em 121 propriedadesde 23 municípios durante os anos de 2011 e 2012. As 1.034 amostras sorológicas de ovinos foramsubmetidas à técnica de Imunodifusão em Gel de Ágar (IDGA), utilizando o kit produzido pelo Institutode Tecnologia do Paraná (TECPAR). Anticorpos anti-Brucella ovis foram observados em 5,37%(70/1.304) dos animais testados, havendo diferença estatística significativa (p < 0,05) para adultos ejovens de ambos os sexos, e entre machos e fêmeas. A prevalência encontrada no Estado do Rio Grandedo Norte, Paraíba e Sergipe foi de 7,66% (36/470), 5,40% (13/241) e 3,54% (21/593), respectivamente.Dos municípios visitados, 91,30% (21/23) eram compostos por rebanhos com sorologia positiva e42,15% (51/121) das propriedades possuíam pelo menos um animal soropositivo. A falta de capacitaçãode trabalhadores (p ≤ 0,05) (odds ratio = 3,68; IC 95% = 1,08-12,60; p = 0, 038) foi identificada comofator de risco. Os resultados obtidos neste trabalho indicam que a infecção por B. ovis encontra-sepresente nos ovinos dos estados do Rio Grande do Norte, Paraíba e Sergipe, sendo necessária a adoçãode medidas sanitárias para evitar a propagação da doença nos rebanhos.(AU)

Diluente Tris suplementado com óleo de Mauritia flexuoxa sobre a qualidade do sêmen caprino após a criopreservação / Tris diluent supplemented with Mauritia flexuoxa oil on the quality of goat semen after cryopreservation

Objetivou-se verificar os efeitos, nos parâmetros espermáticos, na integridade mitocondrial, acrossomal e de membrana em células espermáticas, desencadeados pelo uso do Tris (Tris hidroximetil aminometano) suplementado com óleo de Mauritia flexuoxacomo diluente para criopreservação de sêmen caprino. Quatro caprinos clinicamente saudáveis foram utilizados. Os animais eram alimentados diariamente com volumoso (Pennisetum purpureum, Schum.), concentrado (ração peletizada com teor de 20% proteína, 300 g/animal/dia) e sal mineral específico para Caprinos (Caprinofós®), à vontade. Dois ensaios foram realizados: I ­ Teste de toxicidade; II ­ Criopreservação do sêmen com concentrações ideais. No teste de toxicidade as concentrações avaliadas foram: 5%, 10%, 15% e 20% de diluente a base de óleo de Mauritia flexuoxa. Após o teste de toxicidade, foi escolhido a concentraçãoque apresentou o melhor resultado (5%). Logo após, foram realizadas mais 32 coletas, que foram diluídas em Tris-gema-glicerol (grupo controle) ou diluente contendo óleo vegetal (Mauritia flexuoxa). As amostras foram criopreservadas com auxílio do aparelho Tk3000®. Após o período mínimo de uma semana as palhetas foram descongeladas em banho-maria a 37 °C por 30 segundos, acondicionadas em microtubos de centrifugação e homogeneizadas para a análise imediata de motilidade, vigor espermático e morfologia. Em seguida, por meio de sondas fluorescentes foram avaliadas a integridade de acrossomo, membrana plasmática (Diacetato de Carboxifluresceína e Iodeto de Propídeo) e função mitocondrial sob microscopia de epifluorescência. Quanto a motilidade e vigor, integridade mitocondrial e acrossomal, o grupo buriti foi inferior ao grupo controle. O Tris suplementado com óleo de Mauritia flexuoxa na concentração de 5% não influenciou significativamente a qualidade espermática, porém, observou-se morfologia e integridade de membrana favoráveis. Dessa forma, sendo uma alternativa para substituição de diluentes a base de produtos de origem animal.

The objective was to verify the effects, sperm parameters, mitochondrial, acrosomal and membrane integrity in sperm cells, triggered by the use of Tris (Tris hydroxymethyl aminomethane) supplemented with Mauritia flexuoxa oil as a diluent for cryopreservation of goat semen. Four goats clinically healthy were used. The animals were fed daily with bulky (Pennisetum purpureum, Schum.), concentrate (pelleted feed with 20% protein content, 300 g / animal / day) and mineral salt Specific for Goats (Caprinofós®), ad libitum. Two tests were carried out: I - Toxicity test; II - Semen cryopreservation with ideal concentrations. In the toxicity test as selected were: 5%, 10%, 15% and 20% of Mauritia flexuoxa oil-based diluent. After the toxicity test, the concentration that showed the best result (5%) was chosen. Soon after, a further 32 samples were obtained, which were diluted in Tris-glycerol (control group) or diluent containing vegetable oil (Mauritia flexuoxa). The samples were cryopreserved using the Tk3000® machine. After a minimum of one week, the samples were thawed in a 37 ° C water bath for 30 seconds, packed in centrifugation microtubes and homogenized for immediate analysis of motility, sperm vigor and morphology. Then, by means of fluorescent probes, the integrity of the acrosome, plasma membrane (Carboxyflurescein diacetate and Propidium Iodide) and mitochondrial function under epifluorescence microscopy were evaluated. As for motility and vigor, mitochondrial and acrosomal integrity, the buriti group was inferior to the control group. Tris supplemented with Mauritia flexuoxa oil at a concentration of 5% did not significantly influence sperm quality, however, favorable motility, morphology and membrane integrity were observed. Thus, being an alternative to replace diluents based on products of animal origin.

Epidemiological characterization and risk factors associated with Brucella ovis infection in sheep from the states of Rio Grande do Norte, Paraíba, and Sergipe / Caracterização epidemiológica e fatores de risco associados à infecção por Brucella ovis em ovinos dos estados do Rio Grande do Norte, Paraíba e Sergipe

This study aimed to determine the prevalence of ovine brucellosis in the states of Rio Grande do Norte, Paraíba, and Sergipe. A seroepidemiological survey was carried out in 121 properties under 23 municipalities between 2011 and 2012. The 1,034 serological samples collected from the sheep were subjected to the agar gel immunodiffusion (AGID) test using kits produced by the Paraná Institute of Technology (TECPAR). Anti-Brucella ovis antibodies were observed in 5.37% (70/1,304) of the animals evaluated, and the difference in the incidence of brucellosis between the adults and young animals of both sexes, and between the males and females was statistically significant (p < 0.05). The prevalence of brucellosis in Rio Grande do Norte, Paraíba, and was Sergipe 7.66% (36/470), 5.40% (13/241) and 3.54% (21/593), respectively. Of the municipalities visited, 91.30% (21/23) harbored herds with positive serology, and 42.15% (51/121) of the properties had at least one seropositive animal. The lack of training among the workers (p ? 0.05) (odds ratio = 3.68; 95% CI = 1.08-12.60; p = 0.038) was identified as a risk factor. The results obtained in this study indicate that the sheep from Rio Grande do Norte, Paraíba, and Sergipe are infected by B. ovis, thus necessitating the adoption of proper sanitary measures for preventing the spread of the disease in the herds.

O estudo teve como objetivo determinar a prevalência da Brucelose Ovina nos Estados do Rio Grandedo Norte, Paraíba e Sergipe. Foi realizado o levantamento soroepidemiológico em 121 propriedadesde 23 municípios durante os anos de 2011 e 2012. As 1.034 amostras sorológicas de ovinos foramsubmetidas à técnica de Imunodifusão em Gel de Ágar (IDGA), utilizando o kit produzido pelo Institutode Tecnologia do Paraná (TECPAR). Anticorpos anti-Brucella ovis foram observados em 5,37%(70/1.304) dos animais testados, havendo diferença estatística significativa (p < 0,05) para adultos ejovens de ambos os sexos, e entre machos e fêmeas. A prevalência encontrada no Estado do Rio Grandedo Norte, Paraíba e Sergipe foi de 7,66% (36/470), 5,40% (13/241) e 3,54% (21/593), respectivamente.Dos municípios visitados, 91,30% (21/23) eram compostos por rebanhos com sorologia positiva e42,15% (51/121) das propriedades possuíam pelo menos um animal soropositivo. A falta de capacitaçãode trabalhadores (p ≤ 0,05) (odds ratio = 3,68; IC 95% = 1,08-12,60; p = 0, 038) foi identificada comofator de risco. Os resultados obtidos neste trabalho indicam que a infecção por B. ovis encontra-sepresente nos ovinos dos estados do Rio Grande do Norte, Paraíba e Sergipe, sendo necessária a adoçãode medidas sanitárias para evitar a propagação da doença nos rebanhos.

Taxas de maturação de oócitos congelados lentamente de caprinos / Slowly frozen caprine oocyte maturation rate

The objective was to evaluate the maturation rates of goat oocytes submitted to slow freezing in conventional medium for IVM. For this purpose, cumulus-oocyte complexes aspirated from pubic goat ovaries were classified morphologically and slowly frozen with 1.5 M ethylene glycol. After thawing the cryopreserved oocytes, those classified as viable were matured in conventional in vitro maturation medium. Evaluating the maturation rate, the percentage of matured oocytes in the group that underwent the cryopreservation process is significantly lower (17.6%) when compared to the control group (69.2%), also showing a high percentage of immature oocytes. Several oocyte injuries were found, caused by the studied cryopreservation method, interfering with their oocyte competence. Even with nuclear maturation rates, observed through the extrusion of the first polar corpuscle, the morphologies were altered in most oocytes, and further studies using new techniques and / or other cryoprotectants are necessary.(AU)

Desafios e perspectivas na produção in vivo de embriões de pequenos ruminantes no Nordeste do Brasil / Challenges and perspectives in the in vivo production of small ruminant embryos in northeast of Brazil

Em decorrência da necessidade de aproveitamento de matrizes e reprodutores de alto mérito genético biotecnologias reprodutivas como a Multipla Ovulação e Transferência de Embriões (MOTE) vêm sendo cada vez mais utilizada, garantindo assim a produtividade mesmo diante de obstáculos. Nos últimos anos essa atividade vem apresentando um acentuado desenvolvimento e aprimoramento, principalmente em caprinos e ovinos que se constituem espécies de extrema importância para a Região Nordeste do Brasil, por serem uma das culturas mais vantajosas quando comparado com as demais culturas como a pecuária e por se tratar de uma atividade sustentável do ponto de vista ambiental, sociocultural e econômico. Dentre os principais fatores que ainda afetam o desempenho e sucesso dos programas de Transferência de Embriões (TE) em pequenos ruminantes podemos destacar a seleção de fêmeas doadoras e receptoras, variabilidade de resposta aos protocolos de superovulação, regressão prematura de corpo lúteo (CL), necessidade de etapas cirúrgicas como a laparotomia e laparoscopia e baixa disponibilidade de mão de obra especializada. Nesse sentido, este artigo aborda as principais etapas da produção in vivo de embriões em pequenos ruminantes bem como os desafios e as perspectivas de cada etapa a serem enfrentadas na Região Nordeste do Brasil.(AU)

As a result of the need to use matrices and breeders with high genetic merit, reproductive biotechnologies such as Multiple Ovulation and Embryo Transfer (MOTE) have been increasingly used, thus guaranteeing productivity even in the face of obstacles. In recent years, this activity has shown a marked development and improvement, especially in goats and sheep, which are extremely important species for the Northeast Region of Brazil, as they are one of the most advantageous crops for this region when compared to other crops such as livestock and because it is a sustainable activity from an environmental, socio-cultural and economic point of view. Among the main factors that still affect the performance and success of Embryo Transfer (ET) programs in small ruminants we can highlight the selection of donor and recipient females, variability in response to superovulation protocols, premature corpus luteum (CL) regression, need for surgical steps such as laparotomy and laparoscopy and low availability of specialized labor. In this sense, this article addresses the main stages of invivo embryo production in small ruminants as well as the challenges and perspectives of each stage to be faced in the Northeast Region of Brazil.(AU)

Novos enfoques na avaliação andrológica de bovinos e sua contribuição no melhoramento genético do rebanho / New focuses on andrological evaluation of cattle and its contribution in the genetic improvement of the flock

O processo de fertilização é resultante de diversos mecanismos funcionais e estruturais do espermatozoide. Na andrologia veterinária, no que se refere à avaliação do potencial coeundi e generandi, estão disponíveis diversos testes laboratoriais e de imagem que servem como ferramenta de seleção de touros para a reprodução durante a estação de monta. Estas metodologias auxiliam nos protocolos de criopreservação espermática, na investigação de melhores crioprotetores e diluidores para uso in vivo ou in vitro. No entanto, são poucos utilizadas na rotina diária a campo e restritos aos centros de pesquisa. A presente revisão de literatura objetiva descrever as metodologias que estimam o potencial fecundante do sêmen criopreservado do touro.(AU)

The fertilization process is the result of several functional and structural mechanisms of the sperm. In veterinary andrology, with regard to the evaluation of the coeundi and generandi potential, several laboratory and image tests are available that serve as a tool for selecting bulls for breeding during the breeding season. These methodologies assist in sperm cryopreservation protocols, in the investigation of better cryoprotectants and extenders for use in vivo or in vitro. However, they are few used in the daily routine in the field and restricted to research centers. The present literature review aims to describe the methodologies that estimate the fertile potential of the bulls cryopreserved semen.(AU)

Técnica do cometa para avaliação da integridade do DNA de espermatozoides caprinos criopreservados em ACP-101/102® / Comet technique to assess the DNA integrity of cryopreserved goat sperm in ACP-101/102®

Animal reproduction represents one of the most important factors, and the use of reproductive biotechnologies that help increase production is essential. The comet technique is the simple and quick way to detect pre-mutagenic lesions and assist in studies on environmental biomonitoring, toxicological genetics, biological radiation, DNA repair process and genetic ecotoxicology. The objective of this study was to evaluate the viability of DNA from sperm cells from goat semen submitted to the cryopreservation process in powdered coconut water (ACP101c/102c). The experiment was carried out at the Federal University of Piauí with two goats of reproductive age. The evaluation of the spermatic DNA integrity was performed using an immunofluorescence microscope. Classes from 0 to 3 were used, 0 being no damage and 3, the tail of the comet greater than twice the size of the nucleoid. The results obtained in the G2 test group (ACP 101c-102c) showed a slight tail formation, indicating a slight fragmentation of DNA. It was concluded that in the control group GC1 (TRIS + egg yolk of Gallu gallus domesticus) and experimental group (ACP 101c - 102c) there was no significant difference.(AU)

Avaliação estrutural de espermatozoide caprino criopreservado, através da microscopia eletrônica de transmissão / Ultrastructural evaluation of cryopreserved caprine sperm through transmission electronic microscopy

The ultrastructure evaluation allows the analysis of the sperm cell in a subcellular proportion that is not observed in optical microscopy. Transmission electron microscopy (MET) is a tool used to determine the size and shape of inorganic and biological structures based on the interaction of electrons incident on matter. In this sense, with the use of MET, the objective was to evaluate the ultrastructural changes after the semen manipulation, mainly in the freeze-thaw process, which were not observed in tests using optical microscopy, that is, the morphological integrity of the membranes and goat sperm organelles. The evaluation took place at the Institute of Biosciences of the University of Brasília (UnB). The semen straws were thawed and washed in PBS. Then, they were centrifuged and fixed, contrasted in bloc and subjected to dehydration. The samples were placed for polymerization and ultrathin cuts were made and stored until the time of MET evaluation. In the ultrastructural analyzes by MET in the present work, no harmful actions occurred in either the control or experimental groups. The head regions (plasma membrane and acrosome) remained preserved, with no change in DNA. In conclusion, MET is especially useful tool for cell evaluation.(AU)

Uso da bromelina na qualidade do sêmen caprino congelado/descongelado / Use of bromelain in the quality of frozen/thawed goat sêmen

The objective of this study was to evaluate the effect of bromelain on sperm quality, after defrosting, in goats. For this purpose, five Anglo Nubian goats were used. Semen collection was performed with the aid of an artificial vagina. The semen was evaluated for its macroscopic and microscopic parameters. After the analysis, the total volume of the pool was divided into three groups. One belonging to the control group (ACP-101/102®) and two treatment groups with ACP-101/102® enriched with bromelain in concentrations of 5% and 10%. The samples were cryopreserved with the aid of the Tk3000 device. Then, the straws were immersed in liquid nitrogen and stored in cryogenic cylinders. After one week the samples were thawed and evaluated by the system CASA (Computer Assisted Sperm Analysis). After thawing, the total sperm motility assessed by the system CASA was preserved in the control group and in the treatment group at a concentration of 5%. The curvilinear speed (LCV) and mean trajectory speed (VAP) of sperm were higher in the control group compared to the others. Thus, it is concluded that bromelain does not present genotoxicity to goat sperm, as well as its use is satisfactory in terms of the quality of seminal parameters after thawing.(AU)