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tThis study aimed to compare the accuracy of two diagnostic methods, rectal palpation and transrectalultrasound of the ovaries, and assess the reliability of the diagnosis by palpation method. A total of sixteen (16)dairy farm cows underwent ultrasonographic examinations performed soon after rectal palpation. The accuracyof palpation may seem low, but when we analyze the data, we observed that the main structures diagnosed byultrasound were almost always diagnosed by palpation. Ultrasonography, however, has its strong point in theidentification of complex structures such as hemorrhagic body and follicular cysts. We conclude that ultrasoundas well as rectal palpation can be used as a diagnostic tool, although ultrasound provides good visualization ofovarian structures, allowing their measurement.

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tThis study aimed to compare the accuracy of two diagnostic methods, rectal palpation and transrectalultrasound of the ovaries, and assess the reliability of the diagnosis by palpation method. A total of sixteen (16)dairy farm cows underwent ultrasonographic examinations performed soon after rectal palpation. The accuracyof palpation may seem low, but when we analyze the data, we observed that the main structures diagnosed byultrasound were almost always diagnosed by palpation. Ultrasonography, however, has its strong point in theidentification of complex structures such as hemorrhagic body and follicular cysts. We conclude that ultrasoundas well as rectal palpation can be used as a diagnostic tool, although ultrasound provides good visualization ofovarian structures, allowing their measurement.(AU)

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Microbiological analysis and adulterants researches contribute to surveillance of honey: both ensure trade of the products with sanitary and nutritional quality. The main microorganisms identified in honey are fungi and coliforms, and more frequent adulterations by sugars and syrups of the C4 plants, like sugar cane and corn. In this context, the objective of this work was to investigate the microbiological quality of pure and adulterated honeys marketed in Recife - PE, Brazil. In full, 55 honey samples were submitted to isotopic evaluation honey and its protein, and to search of mold and yeasts; coliforms 35ºC and 45ºC. The results revealed that 31 (56,36%) samples were adulterated with sugars or syrup of the C4 plants and 46 (83,64%) honeys were contaminated with molds and yeasts. Pure and adulterated honeys with sugars or syrups of the C4 plants, contaminated for fungi, were marketed in Recife - PE, Brazil. Its ingestion may harm the consumer when acquire products with microbial contamination.

As análises microbiológicas e as pesquisas de adulterantes contribuem à fiscalização do mel: ambas garantem o comércio de produtos com melhor qualidade sanitária e menor depreciação nutricional. Os principais micro-organismos encontrados no mel são os fungos e coliformes, e as adulterações mais frequentes são por adição de açúcares e xaropes de plantas C4, como os de cana-de-açúcar e milho. Neste contexto, o objetivo deste trabalho foi pesquisar a qualidade microbiológica de méis puros e adulterados comercializados em Recife - PE, Brasil. No total, 55 amostras foram submetidas à avaliação isotópica do mel e de sua proteína, assim como à pesquisa de bolores e leveduras; coliformes 35ºC e 45ºC. Os resultados revelaram que 31 (56,36%) méis estavam adulterados com açúcares ou xaropes provenientes de plantas C4 e 46 (83,64%) apresentaram contaminação por bolores e leveduras. Méis puros e adulterados com açúcares ou xaropes provenientes de plantas C4, contaminados por fungos, são comercializados em Recife - PE, Brasil. Sua ingestão pode causar prejuízos à população consumidora pelo risco de pessoas adquirirem produtos com contaminação microbiana.

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Microbiological analysis and adulterants researches contribute to surveillance of honey: both ensure trade of the products with sanitary and nutritional quality. The main microorganisms identified in honey are fungi and coliforms, and more frequent adulterations by sugars and syrups of the C4 plants, like sugar cane and corn. In this context, the objective of this work was to investigate the microbiological quality of pure and adulterated honeys marketed in Recife - PE, Brazil. In full, 55 honey samples were submitted to isotopic evaluation honey and its protein, and to search of mold and yeasts; coliforms 35ºC and 45ºC. The results revealed that 31 (56,36%) samples were adulterated with sugars or syrup of the C4 plants and 46 (83,64%) honeys were contaminated with molds and yeasts. Pure and adulterated honeys with sugars or syrups of the C4 plants, contaminated for fungi, were marketed in Recife - PE, Brazil. Its ingestion may harm the consumer when acquire products with microbial contamination.(AU)

As análises microbiológicas e as pesquisas de adulterantes contribuem à fiscalização do mel: ambas garantem o comércio de produtos com melhor qualidade sanitária e menor depreciação nutricional. Os principais micro-organismos encontrados no mel são os fungos e coliformes, e as adulterações mais frequentes são por adição de açúcares e xaropes de plantas C4, como os de cana-de-açúcar e milho. Neste contexto, o objetivo deste trabalho foi pesquisar a qualidade microbiológica de méis puros e adulterados comercializados em Recife - PE, Brasil. No total, 55 amostras foram submetidas à avaliação isotópica do mel e de sua proteína, assim como à pesquisa de bolores e leveduras; coliformes 35ºC e 45ºC. Os resultados revelaram que 31 (56,36%) méis estavam adulterados com açúcares ou xaropes provenientes de plantas C4 e 46 (83,64%) apresentaram contaminação por bolores e leveduras. Méis puros e adulterados com açúcares ou xaropes provenientes de plantas C4, contaminados por fungos, são comercializados em Recife - PE, Brasil. Sua ingestão pode causar prejuízos à população consumidora pelo risco de pessoas adquirirem produtos com contaminação microbiana.(AU)

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The objective of this study was to evaluate the effect of biostimulation and temporary weaning on the follicular dynamics and pregnancy rates in Nelore cows. We used three groups of 75 cows: a control group without biostimulation and suckling calves (WB), a group that was biostimulated and had suckling calves (BE) and a group that was biostimulated and subjected to temporary weaning for 56 h (BETW). Ovarian dynamics were examined using ultrasonography. All groups showed follicular atresia. The interval between beginning of the treatment and wave emergence was 3.25 ± 0.30 days for BE, 3.40 ± 0.27 days for BETW and 3.37 ± 0.50 days for WB. The time between implant removal and ovulation was 64.50 ± 1.88 h for BE, 66.75 ± 1.35 h for BETW and 60.85 ± 3.10 h for WB. Eight cows were submitted to ultrasound analysis, and the percentages of cows that had ovulatory follicles of the new follicular wave with maximum diameters greater than 0.80 cm were 100 % (8/8) in BE (1.28 ± 0.12 cm), 100 % (8/8) in BETW (1.52 ± 0.07 cm) and 87.5 % (7/8) in WB (1.21 ± 0.10 cm). The pregnancy rate was 52 % (39/75) in BE, 69.3 % (52/75) in BETW and 37.3 % (28/75) in WB. The association of biostimulation and temporary weaning increased follicular development, ovulation synchronisation and, consequently, the pregnancy rate in beef cows.

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This study was aimed to evaluate breeding seasons of different durations under contrasting climatic conditions upon reproductive performance of nulli­parous does submitted to male effect. A total of 240 Anglo-Nubian females at 8 to 12 months of age were used and were kept away from all three bucks at a distance of 300 m for 60 days, which avoided any physical, visual, olfactory or hearing contact between genders. Estrus were visually detected and pregnancy rates were determined by ultrasonography 60 days after the last mating. Breeding season duration and climatic condition did not influence estrus incidence, where 70% does had estrus detected at the dry season, 80% of does cycled at the rainy season within breeding seasons of 25 days. Moreover, 80% of does cycled at the dry season and 95% at the rainy season during breeding seasons of 35 days and 45 days. After breeding seasons of 25 days, pregnancy rate was lower at the dry season (55.0%) compared to the rainy season (80.0%). Mean birth weight of resulting offspring was lower during the rainy season (2.94 kg ± 0.26) than during the dry season (3.03 kg ± 0.16). The results show that estrus onset is affected by breeding season durations and climatic conditions tested, although dry season lowered pregnancy rates. In conclusion, 25-day breeding seasons are recommended to improve standardization of offspring lots, to reduce costs with food supplementation and estrus detection, while preserving pregnancy and kidding rates.

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This study was aimed to evaluate breeding seasons of different durations under contrasting climatic conditions upon reproductive performance of nulli­parous does submitted to male effect. A total of 240 Anglo-Nubian females at 8 to 12 months of age were used and were kept away from all three bucks at a distance of 300 m for 60 days, which avoided any physical, visual, olfactory or hearing contact between genders. Estrus were visually detected and pregnancy rates were determined by ultrasonography 60 days after the last mating. Breeding season duration and climatic condition did not influence estrus incidence, where 70% does had estrus detected at the dry season, 80% of does cycled at the rainy season within breeding seasons of 25 days. Moreover, 80% of does cycled at the dry season and 95% at the rainy season during breeding seasons of 35 days and 45 days. After breeding seasons of 25 days, pregnancy rate was lower at the dry season (55.0%) compared to the rainy season (80.0%). Mean birth weight of resulting offspring was lower during the rainy season (2.94 kg ± 0.26) than during the dry season (3.03 kg ± 0.16). The results show that estrus onset is affected by breeding season durations and climatic conditions tested, although dry season lowered pregnancy rates. In conclusion, 25-day breeding seasons are recommended to improve standardization of offspring lots, to reduce costs with food supplementation and estrus detection, while preserving pregnancy and kidding rates.(AU)

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The wok was aimed to induce multiple ovulations in mares using low doses of deslorelin acetate (GnRH) to enhance the reproductive efficiency. A total of 15 mares of the quarter horse breed were used as embryo donors in a cross over 15 x 2 experimental design (fifteen animals x two treatments), where animals were monitored for two consecutive cycles. On the control group (CG), the estrous of donors occurred on its natural form and follicular development was monitored by ultrasonography. Ovulation was induced with 2500 IU hCG intravenously, when the follicle reached 35 mm in diameter, moment when artificial insemination was performed. On day 8 (D8) after ovulation, donors were submitted to embryo collection. On Treated Group 2 (TG), donors received, by intramuscular shot, 7.5 mg dinoprost thrometamine and after 48 hours of its administration follicular development was monitored twice a day byultrasound, until the larger follicle reached 23 to 25 mm and the second follicle reached at least 18 mm. Moreover, a super-ovulation protocol was performed with 100 g deslorelin in 12 diâmehour intervals until larger follicle reached 33 to 35 mm. The ovulation was induced with 1000 g deslorelin acetate, by an intramuscular shot, concomitant with 1000 IU hCG intravenously. Artificial insemination was performed immediately after ovulation and embryo collection was on D8. The percentage of mares with multiple ovulations on CG was 13.33% (2/15) and TG of 86.66% (13/15), with diference between groups (P 0.05). The percentage of embryo recovery per flushing was 60% CG and 80% on TG (P<0.05).[...]

Objetivou-se induzir ovulações múltiplas em éguas utilizando baixas doses de acetato de deslorelina (GnRH) visando aumentar a eficiência reprodutiva nessa espécie. Foram utilizadas 15 éguas doadoras de embriões da raça Quarto de Milha, em sistema cross over 15 x 2 (quinze animais x dois tratamentos), no qual cada uma foi monitorada por dois ciclos consecutivos. No Grupo Controle (GC), o estro das doadoras ocorreu de forma natural e o desenvolvimento folicular foi monitorado por ultrassonografia. A ovulação foi induzida com 2500 UI de hCG por via intravenosa, quando o foliculo alcançou diâmetro de 35 mm, momento em que se procedeu a inseminação artificial. No oitavo dia (D8) após ovulação, as doadoras foram submetidas aos procedimentos de colheita embrionária. No Grupo Tratado (GT), as doadoras receberam, por via intramuscular, 7,5 mg de dinoprost-trometamina e após 48 horas de sua administração foi realizado o monitoramento folicular duas vezes ao dia com auxilio de ultrassom, até que o maior folículo atingisse o diâmetro de 23 a 25 mm e o segundo do que 18 mm. Posteriormenteiniciou-se o protocolo de superovulação com a administração de 100 g de GnRH em intervalo de 12 horas até que o maior folículo atingisse o diâmetro entre 33 de 35 mm. A ovulação foi induzida com 1000 µg de GnRH, por via intramuscular, associado a 1000 UI de hCG aplicado por via intravenosa. Imediatamente após a ovulação efetuou-se a inseminação artificial e a colheita de embriões realizada no D8. O percentual de éguas com ovulações múltiplas no GC foi de 13,33% (2/15) e no GT de 86,66% (13/15), havendo diferença (P 0,05). [...]

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RESUMEN

The wok was aimed to induce multiple ovulations in mares using low doses of deslorelin acetate (GnRH) to enhance the reproductive efficiency. A total of 15 mares of the quarter horse breed were used as embryo donors in a cross over 15 x 2 experimental design (fifteen animals x two treatments), where animals were monitored for two consecutive cycles. On the control group (CG), the estrous of donors occurred on its natural form and follicular development was monitored by ultrasonography. Ovulation was induced with 2500 IU hCG intravenously, when the follicle reached 35 mm in diameter, moment when artificial insemination was performed. On day 8 (D8) after ovulation, donors were submitted to embryo collection. On Treated Group 2 (TG), donors received, by intramuscular shot, 7.5 mg dinoprost thrometamine and after 48 hours of its administration follicular development was monitored twice a day byultrasound, until the larger follicle reached 23 to 25 mm and the second follicle reached at least 18 mm. Moreover, a super-ovulation protocol was performed with 100 g deslorelin in 12 diâmehour intervals until larger follicle reached 33 to 35 mm. The ovulation was induced with 1000 g deslorelin acetate, by an intramuscular shot, concomitant with 1000 IU hCG intravenously. Artificial insemination was performed immediately after ovulation and embryo collection was on D8. The percentage of mares with multiple ovulations on CG was 13.33% (2/15) and TG of 86.66% (13/15), with diference between groups (P < 0.05). The mean number of embryos per donor was 1.066±0.497 on TG and 0.6±0.48 on CG (P < 0.05). The percentage of embryos per ovulation was 52.94% on CG and 57.14% on TG, with diference between groups (P < 0.05). The percentage of embryos per flushing was 60.00% on CG and 80.00% on TG, with diference between groups (P > 0.05). The percentage of embryo recovery per flushing was 60% CG and 80% on TG (P<0.05).[...](AU)

Objetivou-se induzir ovulações múltiplas em éguas utilizando baixas doses de acetato de deslorelina (GnRH) visando aumentar a eficiência reprodutiva nessa espécie. Foram utilizadas 15 éguas doadoras de embriões da raça Quarto de Milha, em sistema cross over 15 x 2 (quinze animais x dois tratamentos), no qual cada uma foi monitorada por dois ciclos consecutivos. No Grupo Controle (GC), o estro das doadoras ocorreu de forma natural e o desenvolvimento folicular foi monitorado por ultrassonografia. A ovulação foi induzida com 2500 UI de hCG por via intravenosa, quando o foliculo alcançou diâmetro de 35 mm, momento em que se procedeu a inseminação artificial. No oitavo dia (D8) após ovulação, as doadoras foram submetidas aos procedimentos de colheita embrionária. No Grupo Tratado (GT), as doadoras receberam, por via intramuscular, 7,5 mg de dinoprost-trometamina e após 48 horas de sua administração foi realizado o monitoramento folicular duas vezes ao dia com auxilio de ultrassom, até que o maior folículo atingisse o diâmetro de 23 a 25 mm e o segundo do que 18 mm. Posteriormenteiniciou-se o protocolo de superovulação com a administração de 100 g de GnRH em intervalo de 12 horas até que o maior folículo atingisse o diâmetro entre 33 de 35 mm. A ovulação foi induzida com 1000 µg de GnRH, por via intramuscular, associado a 1000 UI de hCG aplicado por via intravenosa. Imediatamente após a ovulação efetuou-se a inseminação artificial e a colheita de embriões realizada no D8. O percentual de éguas com ovulações múltiplas no GC foi de 13,33% (2/15) e no GT de 86,66% (13/15), havendo diferença (P<0,05). O número médio de embrião por doadora foi de 1,066±0,497 no GT e de 0,6±0,48 no GC, registrando-se diferença (P < 0,05). A porcentagem de embrião por ovulação foi de 52,94% no GC e 57,14% no GT, existindo diferença (P < 0,05). A porcentagem de embrião por lavado foi de 60,00% no GC e de 80,00% no GT, havendo diferença (P > 0,05). [...](AU)

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The objective of this study was to test the “male effect” on the reproductive performance of Anglo Nubian does (n = 180), aged between 24 and 60 months, under different male-to-female ratios (1:20 – T20, 1:30 – T30, and 1:40 – T40) and climatic conditions (dry season – DS, and rainy season – RS). Does were randomly distributed into three groups (T20, T30, and T40) and were isolated from bucks at a distance of 300 m for 60 days before the start of the experiments. The first manifestation of estrous during the DS occurred 6.83 ± 4.54 (T20), 6.72 ± 4.56 (T30) and 7.05 ± 5.23 (T40) days following the onset of the breeding season (P>0.05). In the RS, onset of estrous was observed 6.60 ± 4.74 (T20), 6.70 ± 4.43 (T30) and 7.46 ± 4.54 (T40) days after the beginning of the breeding season (P>0.05). Estrous induction in females during the DS occurred in 95% (T20), 80% (T30), and 75.5% (T40) of all females. During the RS, estrous detection reached 100% (T20), 100% (T30), and 97.5% (T40) of all females, with no difference between all RS and DS groups. Estrous synchronization during the DS occurred in 35.00% (T20), 36.66% (T30), and 32.50% (T40) of all females, for an average occurrence of 34.72%. During the RS, synchronization occurred in 65% (T20), 70% (T30) and 62.25% (T40) of all females, for an average occurrence of 65.75%; no difference was detected between the RS and the DS. Pregnancy rates in the DS groups were 65.0% (T20), 70.0% (T30), and 62.5% (T40), while pregnancy rates in the RS were 90.0% (T20), 86.6% (T30), and 95.0% (T40). No difference was observed for conception rates between any of the RS and DS groups. Prolificacy during the DS was 1.30 (T20), 1.30 (T30) and 1.35 (T40), while in the RS prolificacy was 1.29 (T20), 1.25 (T30) and 1.30 (T40). Thus, the male effect can be used effectively for goats under 1:20–1:40 male-to-female ratios in a 45-day mating season under varying climatic conditions.

O objetivo do presente trabalho foi testar o “efeito macho” sobre o desempenho reprodutivo de cabras Anglo Nubianas (n = 180), com idade entre 24 e 60 meses, sob diferentes proporções macho:fêmea (1:20 - T20, 1:30 - T30, e 01:40 - T40) e condições climáticas (estação seca - ES, e estação chuvosa - EC). As fêmeas foram distribuídas aleatoriamente em três grupos (T20, T30 e T40), e foram mantidas separadas dos machos a distância de 300 m por 60 dias antes do início do experimento. A primeira manifestação de estro na ES ocorreu 6,83 ± 4,54 (T20), 6,72 ± 4,56 (T30) e em 7,05 ± 5,23 (T40) dias após o início da época de reprodução (P>0,05). Na EC, o início do estro foi observado 6,60 ± 4,74 (T20), 6,70 ± 4,43 (T30) e 7,46 ± 4,54 (T40) dias após o início da época de reprodução (P>0,05). A indução de estro na ES chegou em 95% (T20), 80% (T30), e 75,5% (T40) das fêmeas. Na EC, a detecção do estro atingiu 100% (T20), 100% (T30), e 97,5% (T40), sem diferença entre as épocas do ano. Sincronização de estro durante ES ocorreu em 34,72%, 35,00% (T20), 36,66% (T30), e 32,5 % (T40) das fêmeas. Na EC, a sincronização ocorreu em 65,75% das fêmeas, 65% (T20), 70% (T30) e 62,25% (T40), sem apresentar diferença estatística (P>0,05) entre EC e ES. A taxa de gestação nos grupos ES foi de 65,0% (T20), 70,0% (T30), e 62,5% (T40), enquanto que em EC foi de 90,0% (T20), 86,6% (T30), e de 95,0% (T40), sem diferença estatística (P>0,05) entre EC e ES. A prolificidade durante ES foi 1,30 (T20), 1,30 (T30) e 1,35 (T40) e na EC foi 1,29 (T20), 1,25 (T30) e 1,30 (T40). Em conclusão, o efeito macho pode ser utilizado com as proporções 1:20-1:40 macho:fêmea, em estação de monta de 45 dias de duração sob condições climáticas diferentes.

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The objective of this study was to test the “male effect” on the reproductive performance of Anglo Nubian does (n = 180), aged between 24 and 60 months, under different male-to-female ratios (1:20 T20, 1:30 T30, and 1:40 T40) and climatic conditions (dry season DS, and rainy season RS). Does were randomly distributed into three groups (T20, T30, and T40) and were isolated from bucks at a distance of 300 m for 60 days before the start of the experiments. The first manifestation of estrous during the DS occurred 6.83 ± 4.54 (T20), 6.72 ± 4.56 (T30) and 7.05 ± 5.23 (T40) days following the onset of the breeding season (P>0.05). In the RS, onset of estrous was observed 6.60 ± 4.74 (T20), 6.70 ± 4.43 (T30) and 7.46 ± 4.54 (T40) days after the beginning of the breeding season (P>0.05). Estrous induction in females during the DS occurred in 95% (T20), 80% (T30), and 75.5% (T40) of all females. During the RS, estrous detection reached 100% (T20), 100% (T30), and 97.5% (T40) of all females, with no difference between all RS and DS groups. Estrous synchronization during the DS occurred in 35.00% (T20), 36.66% (T30), and 32.50% (T40) of all females, for an average occurrence of 34.72%. During the RS, synchronization occurred in 65% (T20), 70% (T30) and 62.25% (T40) of all females, for an average occurrence of 65.75%; no difference was detected between the RS and the DS. Pregnancy rates in the DS groups were 65.0% (T20), 70.0% (T30), and 62.5% (T40), while pregnancy rates in the RS were 90.0% (T20), 86.6% (T30), and 95.0% (T40). No difference was observed for conception rates between any of the RS and DS groups. Prolificacy during the DS was 1.30 (T20), 1.30 (T30) and 1.35 (T40), while in the RS prolificacy was 1.29 (T20), 1.25 (T30) and 1.30 (T40). Thus, the male effect can be used effectively for goats under 1:20–1:40 male-to-female ratios in a 45-day mating season under varying climatic conditions.(AU)

O objetivo do presente trabalho foi testar o “efeito macho” sobre o desempenho reprodutivo de cabras Anglo Nubianas (n = 180), com idade entre 24 e 60 meses, sob diferentes proporções macho:fêmea (1:20 - T20, 1:30 - T30, e 01:40 - T40) e condições climáticas (estação seca - ES, e estação chuvosa - EC). As fêmeas foram distribuídas aleatoriamente em três grupos (T20, T30 e T40), e foram mantidas separadas dos machos a distância de 300 m por 60 dias antes do início do experimento. A primeira manifestação de estro na ES ocorreu 6,83 ± 4,54 (T20), 6,72 ± 4,56 (T30) e em 7,05 ± 5,23 (T40) dias após o início da época de reprodução (P>0,05). Na EC, o início do estro foi observado 6,60 ± 4,74 (T20), 6,70 ± 4,43 (T30) e 7,46 ± 4,54 (T40) dias após o início da época de reprodução (P>0,05). A indução de estro na ES chegou em 95% (T20), 80% (T30), e 75,5% (T40) das fêmeas. Na EC, a detecção do estro atingiu 100% (T20), 100% (T30), e 97,5% (T40), sem diferença entre as épocas do ano. Sincronização de estro durante ES ocorreu em 34,72%, 35,00% (T20), 36,66% (T30), e 32,5 % (T40) das fêmeas. Na EC, a sincronização ocorreu em 65,75% das fêmeas, 65% (T20), 70% (T30) e 62,25% (T40), sem apresentar diferença estatística (P>0,05) entre EC e ES. A taxa de gestação nos grupos ES foi de 65,0% (T20), 70,0% (T30), e 62,5% (T40), enquanto que em EC foi de 90,0% (T20), 86,6% (T30), e de 95,0% (T40), sem diferença estatística (P>0,05) entre EC e ES. A prolificidade durante ES foi 1,30 (T20), 1,30 (T30) e 1,35 (T40) e na EC foi 1,29 (T20), 1,25 (T30) e 1,30 (T40). Em conclusão, o efeito macho pode ser utilizado com as proporções 1:20-1:40 macho:fêmea, em estação de monta de 45 dias de duração sob condições climáticas diferentes.(AU)

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Neste trabalho foi avaliado o efeito do estresse calórico durante a maturação de oócitos sobre a produção in vitro de embriões ovinos. Os ovários foram obtidos em abatedouro e os oócitos colhidos de folículos de 2 a 6 mm de diâmetro. Após seleção, os oócitos, em 10 replicações, foram colocados para maturação in vitro (MIV) durante 24 horas. Os oócitos submetidos ao estresse térmico de 41º C durante 3, 6, 12, 18 e 24 horas foram posteriormente transferidos para completar a MIV a 39 ºC, mesma temperatura utilizada para maturação dos oócitos do grupo controle. O desenvolvimento dos embriões foi determinado nos dias 3, 4, 5 e 8 pós-fecundação. A avaliação da qualidade dos embriões foi efetuada através da contagem total de células coradas pelo DAPI e da determinação do número de blastômeros positivos para apoptose através do teste de TUNEL. Observou-se que o estresse térmico diminuiu (P 0,05) somente até 18 horas de incubação. Os resultados permitem concluir que o estresse calórico durante a maturação in vitro de oócitos reduz a quantidade e a qualidade dos embriões ovinos produzidos in vitro determinadas pela alta incidência de apoptose.

The aim of this work was evaluate the effect of heat stress during oocyte maturation on ovine in vitro embryo production. Ovaries were collected at abattoirs and oocytes retrieved from follicles ranging from 2 and 6 mm in diameter. After selection, all oocytes, in 10 replicates, were placed in in vitro maturation (IVM) during 24 hours. The oocytes were submitted to heat stress of 41º C during 3, 6, 12, 18 and 24 hours and were further transferred to 39º C in order to complete IVM, which was the temperature of maturation of control oocytes. Embryonic development was determined on days 3, 4, 5, and 8 post-fertilization. Embryo evaluation was performed as total cell count by DAPI staining and determination of positive blastomere for apoptosis by the TUNEL assay. We observed that heat stress diminishes (P 0.05) only up to 18 hours of incubation. The results allow the conclusion that heat stress during oocyte in vitro maturation reduces the quantity and quality of ovine embryos produced in vitro determined by the high incidence of apoptosis.

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Neste trabalho foi avaliado o efeito do estresse calórico durante a maturação de oócitos sobre a produção in vitro de embriões ovinos. Os ovários foram obtidos em abatedouro e os oócitos colhidos de folículos de 2 a 6 mm de diâmetro. Após seleção, os oócitos, em 10 replicações, foram colocados para maturação in vitro (MIV) durante 24 horas. Os oócitos submetidos ao estresse térmico de 41º C durante 3, 6, 12, 18 e 24 horas foram posteriormente transferidos para completar a MIV a 39 ºC, mesma temperatura utilizada para maturação dos oócitos do grupo controle. O desenvolvimento dos embriões foi determinado nos dias 3, 4, 5 e 8 pós-fecundação. A avaliação da qualidade dos embriões foi efetuada através da contagem total de células coradas pelo DAPI e da determinação do número de blastômeros positivos para apoptose através do teste de TUNEL. Observou-se que o estresse térmico diminuiu (P < 0,05) a capacidade de maturação dos oócitos de acordo com o tempo de exposição à temperatura de 41º C. No grupo de oócitos incubados a 39 C, 70,70% maturou, enquanto que nos grupos expostos ao estresse térmico, apenas 45,28%, 35,17%, 12,30%, 9,74% e 4,60% maturaram, respectivamente, após 3, 6, 12, 18 e 24 horas de incubação. A duração de exposição dos oócitos ao estresse calórico é inversamente proporcional (P < 0,05) à capacidade de desenvolvimento embrionário e diretamente proporcional (P < 0,05) ao número de blastocistos positivos para apoptose. Todavia, o efeito deletério do estresse térmico sobre a clivagem e os embriões nos estádios de 8 a 16 células e de mórula foi crescente (P > 0,05) somente até 18 horas de incubação. Os resultados permitem concluir que o estresse calórico durante a maturação in vitro de oócitos reduz a quantidade e a qualidade dos embriões ovinos produzidos in vitro determinadas pela alta incidência de apoptose.(AU)

The aim of this work was evaluate the effect of heat stress during oocyte maturation on ovine in vitro embryo production. Ovaries were collected at abattoirs and oocytes retrieved from follicles ranging from 2 and 6 mm in diameter. After selection, all oocytes, in 10 replicates, were placed in in vitro maturation (IVM) during 24 hours. The oocytes were submitted to heat stress of 41º C during 3, 6, 12, 18 and 24 hours and were further transferred to 39º C in order to complete IVM, which was the temperature of maturation of control oocytes. Embryonic development was determined on days 3, 4, 5, and 8 post-fertilization. Embryo evaluation was performed as total cell count by DAPI staining and determination of positive blastomere for apoptosis by the TUNEL assay. We observed that heat stress diminishes (P < 0.05) oocyte maturation capacity in accordance with exposure time of 41º C. In the group of oocytes incubated at 39 ºC, 70.70% matured, while in the groups exposed to heat stress of 41º C, only 45.28%, 35.17%, 12.30%, 9.74% and 4.60% matured, respectively, after 3, 6, 12, 18 and 24 hours of incubation. The duration of exposure to heat stress is inversely proportional (P < 0.05) to embryonic developmental capacity directly proportional (P < 0.05) to the number of blastocysts positive to apoptosis. However, the cleavage rate and embryonic development from 8 to 16 cells and morulae stages were affected by heat stress (P > 0.05) only up to 18 hours of incubation. The results allow the conclusion that heat stress during oocyte in vitro maturation reduces the quantity and quality of ovine embryos produced in vitro determined by the high incidence of apoptosis.(AU)

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Objetivou-se com este estudo determinar a influência das estações seca e chuvosa na maturação de oócitos e produção in vitro (PIV) de embriões na espécie caprina. Os ovários das cabras nas estações seca (outubro a março) e chuvosa (abril a setembro) foram colhidos em abatedouro e transportados ao Laboratório de Biotécnicas da Reprodução da UFRPE. Os complexos cumulus oophorus (CCOs) foram colhidos pela técnica de "slicing" dos folículos entre 2 a 6 mm de diâmetro e selecionados com base na classificação morfológica. Foram realizadas 12 repetições, nas quais os CCOs foram submetidos à maturação, fertilização e cultivo in vitro dos embriões. A média e desvio padrão da taxa de clivagem foi determinada no dia 3 (D-3) e dos embriões que se desenvolveram aos estádios de 8-16 células, mórula e blastocisto foi determinada nos dias 4 (D-4), 5 (D-5) e 8 (D-8) após a fecundação, respectivamente. A quantidade de blastômeros foi determinada com o corante DAPI e os blastômeros positivos para apoptose por meio do teste de TUNEL. A produção de embriões no D-3 e mórulas foram inferiores ao obtido no período chuvoso (P 0,05) quanto às fases de maturação, fertilização, cultivo no D-4 e blastocisto. Os embriões produzidos na estação seca apresentaram maior incidência de apoptose (P < 0,05). Nas condições descritas neste estudo, os resultados permitem concluir que as fases iniciais do desenvolvimento embrionário sofrem maior impacto negativo durante a estação seca em protocolos de PIV na espécie caprina.

This study aimed to determine the influence of dry and rainy seasons on oocyte maturation and in vitro production (IVP) of embryos in goats. The ovaries of does in dry (October to March) and rainy season (April-September) were collected at a slaughterhouse and transported to the Laboratory of Biotechnical Reproduction of UFRPE. The cumulus oophorus complexes (COCs) were collected by the technique of slicing of the follicles from 2 to 6 mm in diameter, and selected based on morphologic classification. We used 12 replicates, in which the COCs were submitted to maturation, fertilization and in vitro culture. The cleavage rate was determined on day 3 (D-3) and embryos that developed to 8-16 cells (D-4), morulae (D-5) and blastocyst (D-8) stages after fertilization. The number of blastomeres was assessed with DAPI staining, and the determination of apoptotic blastomeres was performed by TUNEL assay. In the dry season, D-3 embryos and morulae development was lower than that obtained in the rainy season (P0.05) were observed on oocyte maturation, fertilization, D-4 embryo yield and blastocyst development. Embryos produced during the dry season had a higher incidence of apoptosis on D- 3 and at the morulae stage (P<0.05). Under the conditions described in this study, the results suggest that the early stages of embryonic development suffer greater negative impact during the dry season in IVP protocols in goats.

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Objetivou-se com este estudo determinar a influência das estações seca e chuvosa na maturação de oócitos e produção in vitro (PIV) de embriões na espécie caprina. Os ovários das cabras nas estações seca (outubro a março) e chuvosa (abril a setembro) foram colhidos em abatedouro e transportados ao Laboratório de Biotécnicas da Reprodução da UFRPE. Os complexos cumulus oophorus (CCOs) foram colhidos pela técnica de "slicing" dos folículos entre 2 a 6 mm de diâmetro e selecionados com base na classificação morfológica. Foram realizadas 12 repetições, nas quais os CCOs foram submetidos à maturação, fertilização e cultivo in vitro dos embriões. A média e desvio padrão da taxa de clivagem foi determinada no dia 3 (D-3) e dos embriões que se desenvolveram aos estádios de 8-16 células, mórula e blastocisto foi determinada nos dias 4 (D-4), 5 (D-5) e 8 (D-8) após a fecundação, respectivamente. A quantidade de blastômeros foi determinada com o corante DAPI e os blastômeros positivos para apoptose por meio do teste de TUNEL. A produção de embriões no D-3 e mórulas foram inferiores ao obtido no período chuvoso (P < 0,05). Não apresentaram diferenças (P > 0,05) quanto às fases de maturação, fertilização, cultivo no D-4 e blastocisto. Os embriões produzidos na estação seca apresentaram maior incidência de apoptose (P < 0,05). Nas condições descritas neste estudo, os resultados permitem concluir que as fases iniciais do desenvolvimento embrionário sofrem maior impacto negativo durante a estação seca em protocolos de PIV na espécie caprina.(AU)

This study aimed to determine the influence of dry and rainy seasons on oocyte maturation and in vitro production (IVP) of embryos in goats. The ovaries of does in dry (October to March) and rainy season (April-September) were collected at a slaughterhouse and transported to the Laboratory of Biotechnical Reproduction of UFRPE. The cumulus oophorus complexes (COCs) were collected by the technique of slicing of the follicles from 2 to 6 mm in diameter, and selected based on morphologic classification. We used 12 replicates, in which the COCs were submitted to maturation, fertilization and in vitro culture. The cleavage rate was determined on day 3 (D-3) and embryos that developed to 8-16 cells (D-4), morulae (D-5) and blastocyst (D-8) stages after fertilization. The number of blastomeres was assessed with DAPI staining, and the determination of apoptotic blastomeres was performed by TUNEL assay. In the dry season, D-3 embryos and morulae development was lower than that obtained in the rainy season (P<0.05). However, no differences (P>0.05) were observed on oocyte maturation, fertilization, D-4 embryo yield and blastocyst development. Embryos produced during the dry season had a higher incidence of apoptosis on D- 3 and at the morulae stage (P<0.05). Under the conditions described in this study, the results suggest that the early stages of embryonic development suffer greater negative impact during the dry season in IVP protocols in goats.(AU)

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Background: The low productivity of Northeast goat herds has been circumvented by the importation of exotic species; however, caution is needed due to the susceptibility of these breeds to the high temperatures found in this region. It is now known that the oocyte and the embryo are the primary targets of the deleterious effects induced by heat stress, causing cellular damage that triggers the cascade of apoptosis. Therefore, the purpose of this study was to evaluate the effect of thermal heat stress during in vitro maturation of oocytes and its effects on embryo production in goats. Materials, Methods & Results: The ovaries were collected in a slaughterhouse and transported to the Laboratory of Biotechnical Reproduction of UFRPE. The cumulus oophorus complexes (COCs) were collected by the technique of slicing the follicles from 2 to 6 mm in diameter, selected based on morphology and placed in a basic medium for maturation. In 10 replications, the COCs were submitted to the thermal heat stress at 41°C for 0 (thermoneutrality at 39°C), 3, 6, 12, 18, and 24 h of maturation in vitro. The data was evaluated in maturation, fertilization, cleavage (D-3), stage of 8-16 cells (D4), morula (D-5), and blastocyst (D-8) after fertilization and blastocysts positive for apoptosis through the TUNEL test. For statistics, the results were expressed as mean and standard deviation. Also considering the measurements addressed in percentages, a comparison of variances was carried out, F-test for variances to the level of signifi cance 5% (P < 0.05). Then, a t-test to compare averages was conducted, to the significance level of 5%, for equivalent variances or distinct variances, according to what was observed in the F-test for variances. A signifi cant difference (P < 0.05) was observed during all time periods studied for heat stress on maturation, fertilization, D-3, D-4, and D-5. On D-8 no significant difference (P > 0.05) was observed between the periods of 3 vs 6 and 18 vs 24 h, and in the blastocysts positive by the TUNEL test for the periods of 0 vs 3, 3 vs 6, 12 vs 18, and 18 vs 24 h of heat stress. Discussion: When applying a thermal shock that produces damage to the oocyte maturation in vitro, the characteristic membrane, chromatin configuration, and meiotic spindles are changed, and thus, the developmental potential of oocytes after fertilization is compromised. It was observed in this study that there was a gradual reduction in the number of oocytes as the time of exposure to heat shock increased, reflecting directly on each stage of IVP embryos. These stages are most vulnerable during maturation in vivo (ovulation), fertilization, within two days after fertilization, and in the first division of cleavage, as evidenced in this study in vitro after heat stress, reducing the number of blastocysts. This suggests that apoptosis can be induced in pre-implantation of embryos exposed to maternal hyperthermia. Moreover, the degree of apoptosis experienced by IVP embryos generally reflects the severity of thermal shock. In this study, the percentage of cells that were TUNEL positive increased with the prolongation of thermal shock. Induction of apoptosis was time dependent and the number of apoptotic cells increased proportionally after 6, 12, 18, and 24 h of exposure. Under the conditions observed in this study, the results indicate that the time in which the oocyte is exposed to heat stress during maturation in vitro is of great importance for embryonic development and their level of apoptotic cells.

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The objective of this study was to identify the migration period of the genital tubercle and the period of visualization of external genital structures in fetuses of the Dorper breed of sheep derived from natural mating and from fresh, frozen and vitrified embryo transfer. Transrectal ultrasound was performed using a double-frequency linear transducer (6.0 and 8.0 MHz) to monitor 130 ewe fetuses distributed in the four treatments regarding embryo origin. The accuracy of the ultrasound was 100% in this experiment. The fetuses originated from controlled natural mating (NM) and from fresh (FrE), frozen (FE) and vitrified (VE) embryo transfer, with embryos collected 7 days after breeding. Migration of the genital tubercle occurred earlier (P<0.05) in NM (42.21+/-2.86 days) than in FrE (43.98+/-3.00 days), FE (44.97+/-1.83 days) and VE (44.58+/-1.97 days). Visualization of the scrotal bag, prepuce and vulva occurred, respectively, earlier (P<0.05) in NM (45.22+/-1.25, 45.95+/-1.53 and 45.01+/-1.03 days) than in FrE (48.91+/-1.92, 48.52+/-1.41 and 47.41+/-1.41 days), FE (49.97+/-1.08, 49.18+/-2.00 and 47.64+/-1.82 days) and VE (50.12+/-1.66, 49.27+/-1.61 and 47.93+/-1.92 days). The results show that fetal sexing can be accomplished from the 50th day onward in fetuses produced by natural mating and from the 55th day onward in fetuses derived from fresh, frozen and vitrified embryos. It can also be concluded that real-time ultrasonography is a reliable tool for fetal sex determination in sheep taking into account both the location of the genital tubercle and the identification of external genital structures.

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Procurou-se estabelecer o período para sexar fetos caprinos pela ultrassonografia transretal visibilizando-se as estruturas da genitália externa, sendo que para tal foi monitorado o dia do posicionamento final do tubérculo genital (TG), bem como o dia em que o pênis, a bolsa escrotal, o clitóris e as tetas foram visibilizados. Foram monitorados fetos das raças Boer (n = 36), Parda Alpina (n = 31) e Anglo-nubiana (n = 27), do 40º ao 60º dia de gestação, usando-se transdutor linear de 6,0 e 8,0 MHz. O posicionamento final do TG ocorreu no período de 47,11 ± 1,45 dias, nos machos, e 45,62 ± 1,36 dias, nas fêmeas, e a visibilização das estruturas da genitália externa no período de 49,42 ± 2,20 dias para a bolsa escrotal; 49,37 ± 2,19 dias para o pênis; 49,23 ± 1,75 dias para as tetas e 49,98 ± 2,52 dias para o clitóris. A migração do TG no feto fêmea foi mais precoce (P 0,05) entre as estruturas referentes ao mesmo sexo. Conclui-se que apesar da sexagem de fetos caprinos poder ser efetuada antes do 55º dia de gestação, recomenda-se realizá-la somente após a visibilização das estruturas da genitália externa para uma maior segurança na identificação do sexo.

The aim of this study was to establish the period for sexing caprine fetuses by transrectal ultrasonography through viewing the structures of the external genitalia, monitoring of the final positioning of the genital tubercle, and the day that the penis, scrotum, clitoris and nipples were seen. Fetuses of Boer (n = 36), Brownscale Alpine (n = 31) and Anglo-nubian (n = 27), breeds in the 40th-60th day of gestation, were tracked using linear transducer of 6.0 and 8.0 MHz. The final positioning of the genital tubercle occurred in the period from 47.11 ± 1.45 days in males and 45.62 ± 1.36 days in females. The visualization of structures of the external genitalia was in the period from 49.42 ± 2.20 days for scrotum; 49.37 ± 2.19 days for penis; 49.23 ± 1.75 days for nipples and 49.98 ± 2.52 days for clitoris. The migration of female genital tubercle in the fetus occurred earlier (P 0.05) among the structures of the same sex. In conclusion, although the sexing of caprine fetuses can be done before the 55th day of pregnancy, it is recommended to perform it only after the visualization of the structures of the external genitalia for a greater security in the sex identification.

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O presente estudo teve como objetivo avaliar a influência do diâmetro folicular sobre a qualidade dos oócitos obtidos de ovários de fêmeas ovinas e caprinas. Utilizaram-se 156 ovários de ovelhas e 105 de cabras, provenientes de abatedouro. Os folículos foram mensurados, aspirados e divididos em três classes de diâmetro folicular: classe 1 (CI) – 2 a 3 mm, classe 2 (CII) – 4 a 5 mm e classe 3 (CIII) – 6 mm. Os oócitos recuperados foram avaliados e classificados quanto ao aspecto morfológico em cinco grupos de qualidade: grau 1 (GI), grau 2 (GII), grau 3 (GIII), desnudo (D) e atrésicos (A). Dos 468 folículos aspirados de ovinos, foram encontrados 327 CI, 84 CII, 57 CIII e 83 GI, 78 GII, 95 GIII, 119 D, 93 A. E dos 422 folículos aspirados de caprinos, foram encontrados 197 CI, 132 CII, 92 CIII e 64 GI, 70 GII, 91 GIII, 123 D, 74 A. Os dados obtidos demonstram não haver correlações significativas entre qualidade dos oócitos e diâmetro dos folículos (P > 0,05). Nas condições em que se desenvolveu este estudo, os resultados permitem concluir que o diâmetro folicular não exerce influência sobre a qualidade do complexo cumulus-oócito (COC) recuperado de ovários de fêmeas ovinas e caprinas.

This study aimed to evaluate the influence of follicular diameter on the quality of oocytes from ovaries of female sheep and goats. The ovaries of sheep (156 units) and goats (105 units) from slaughterhouse were used. The follicles were measured, aspirated and divided into three classes of follicular diameter: class 1 (CI) – 2 to 3 mm, class 2 (CII) – 4 to 5 mm and class 3 (CIII) – 6 mm. The recovered oocytes were evaluated and classified according to morphological aspect in 5 quality groups: grade 1 (GI), grade 2 (GII), grade 3 (GIII), naked (N) and atresia (A). Of the 468 follicles aspirated from sheep, 327 CI, 84 CII, CIII 57 and 83 GI, 78 GII, GIII 95, 119 N, 93 A were found, and from the 422 follicles aspirated from goats 197 CI, 132 CII, 92 CIII and 64 GI, 70 GII, 91 GIII, 123 N, 74 A were found. The data did not show significant correlation between quality of oocytes and diameter of follicles (P > 0.05). Under the conditions observed in this study it was concluded that the follicular diameter has no influence on the quality of cumulus-oocyte complex (COC) recovered from the ovaries of female sheep and goats.

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O presente estudo teve como objetivo avaliar a influência do diâmetro folicular sobre a qualidade dos oócitos obtidos de ovários de fêmeas ovinas e caprinas. Utilizaram-se 156 ovários de ovelhas e 105 de cabras, provenientes de abatedouro. Os folículos foram mensurados, aspirados e divididos em três classes de diâmetro folicular: classe 1 (CI) 2 a 3 mm, classe 2 (CII) 4 a 5 mm e classe 3 (CIII) 6 mm. Os oócitos recuperados foram avaliados e classificados quanto ao aspecto morfológico em cinco grupos de qualidade: grau 1 (GI), grau 2 (GII), grau 3 (GIII), desnudo (D) e atrésicos (A). Dos 468 folículos aspirados de ovinos, foram encontrados 327 CI, 84 CII, 57 CIII e 83 GI, 78 GII, 95 GIII, 119 D, 93 A. E dos 422 folículos aspirados de caprinos, foram encontrados 197 CI, 132 CII, 92 CIII e 64 GI, 70 GII, 91 GIII, 123 D, 74 A. Os dados obtidos demonstram não haver correlações significativas entre qualidade dos oócitos e diâmetro dos folículos (P > 0,05). Nas condições em que se desenvolveu este estudo, os resultados permitem concluir que o diâmetro folicular não exerce influência sobre a qualidade do complexo cumulus-oócito (COC) recuperado de ovários de fêmeas ovinas e caprinas.(AU)

This study aimed to evaluate the influence of follicular diameter on the quality of oocytes from ovaries of female sheep and goats. The ovaries of sheep (156 units) and goats (105 units) from slaughterhouse were used. The follicles were measured, aspirated and divided into three classes of follicular diameter: class 1 (CI) 2 to 3 mm, class 2 (CII) 4 to 5 mm and class 3 (CIII) 6 mm. The recovered oocytes were evaluated and classified according to morphological aspect in 5 quality groups: grade 1 (GI), grade 2 (GII), grade 3 (GIII), naked (N) and atresia (A). Of the 468 follicles aspirated from sheep, 327 CI, 84 CII, CIII 57 and 83 GI, 78 GII, GIII 95, 119 N, 93 A were found, and from the 422 follicles aspirated from goats 197 CI, 132 CII, 92 CIII and 64 GI, 70 GII, 91 GIII, 123 N, 74 A were found. The data did not show significant correlation between quality of oocytes and diameter of follicles (P > 0.05). Under the conditions observed in this study it was concluded that the follicular diameter has no influence on the quality of cumulus-oocyte complex (COC) recovered from the ovaries of female sheep and goats.(AU)

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