The present study evaluated the effects of heat stress on the ram seminal plasma proteome. Six Morada Nova rams were scrotal insulated for 8 days. Scrotal circumference, sperm parameters, and seminal fluid proteins were evaluated before (Day 0) and twice during scrotal insulation (Days 4 and 8), and weekly until semen parameters returned to preinsulation values (normal). Seminal proteins were analyzed by two-dimensional SDS-PAGE and mass spectrometry. Scrotal circumference decreased from 30 ± 0.4 cm on Day 0 to 22.6 ± 0.6 cm on Day 36 (P < 0.05) and became equivalent to preinsulation values on Day 71. Motile sperm became nearly absent from Day 8 to Day 64 but returned to normal on Day 113. Percentage of normal sperm changed similarly and returned to normal on Day 106. Rams were azoospermic between Days 29 and 64, and sperm concentration came back to normal on Day 92. The number of spots/two-dimensional gel reduced from 256 ± 31 on Day 0 to 104 ± 14 on Day 29 (when rams were azoospermic) and then increased to 183 ± 9 on Day 113 (P < 0.05), similar to spot counts before insulation. The intensities of 24 spots, referring to 17 seminal plasma proteins, were affected by treatment (P < 0.05). After insulation, seminal plasma had greater expression of actin (two isoforms), albumin, heat shock protein 70 kDa, protein DJ-1, HRPE773-like, C-reactive protein precursor, bodhesin-2 (one isoform), spermadhesins. Most protein spots had the greatest intensity between Days 8 and 29, returning to preinsulation values on Day 113 (when many sperm criteria returned to normal). Proteins downregulated after scrotal insulation included dipeptidyl peptidase 3, isoforms of heat shock protein 90 kDa, RSVP22, MMP2 and of Bdh2. In this case, RSVP22 was reduced on Day 113 and all others, on Day 134. Expression of MMP2 and HSP90.1 was reduced throughout the study. Integrin β5, V-type H(+)-ATPase subunit A, ZBTB 42-like protein, isoforms of Bdh2, PSP-I, and RSVP22 were upregulated after testis insulation. Intensities of these spots were maximum (P < 0.05) 8 days after insulation started or on Day 29. Expression of most of such proteins returned to normal on Day 113. In conclusion, scrotal insulation affected testis and sperm parameters of rams, indicating alterations in both spermatogenesis and sperm maturation. Changes of seminal plasma proteome were coincidental with variations in semen parameters. Proteins affected by heat challenge are potentially involved in sperm protection, maturation, and fertilization.

Resumo -O objetivo deste trabalho foi avaliar o efeito de um sistema silvipastoril no conforto térmico de 20 búfalas Murrah, das quais 10 criadas em piquetes sem sombra (SS) e 10 com sombreamento (CS) de Racosperma mangium, em Belém, PA. Os animais foram alimentados em pasto, com Urochloa humidicola, com acesso livre à água para beber e sal mineral. A cada três dias, foram mensuradas: temperatura do ar (TA), umidade relativa do ar (UR), temperatura de globo negro (TGN), temperatura retal (TR), frequências respiratória (FR) e cardíaca (FC), e a temperatura da superfície corporal (TSC), pela manhã (7h) e à tarde (13h). Os valores de TR, TSC, FR e FC foram maiores à tarde, especialmente no grupo SS. Mais altas no período menos chuvoso, a TR, TSC e FR apresentaram correlação linear positiva com a TA e o índice de temperatura e umidade (ITGU) e negativa com a UR. Tanto na estação mais chuvosa quanto na menos chuvosa, a FC apresentou correlações significativas positivas com a TA e ITGU e negativas com a UR, apenas no período mais chuvoso. A arborização da pastagem é eficiente para melhorar o conforto térmico das búfalas Murrah, principalmente à tarde.Termos para indexação: índice de temperatura e umidade, somreamento, bioclimatologia, Bubalus bubalis, Racosperma mangium, fisiologia. Thermal comfort of female buffaloes in a silvipasture system in the eastern AmazonAbstract -The objective of this work was to evaluate the effects of a silvipasture system on the thermal comfort of 20 female buffaloes, from which 10 raised in under nonshaded paddocks (SS) and 10 in paddocks shaded by Racosperma mangium (CS), in Belém county, PA, Brazil. All animals were fed in pastures with Urochloa humidicola, and had free access to drinking water and mineral salt. Every three days, measurements were made for air temperature (TA), relative humidity (UR), black globe temperature (TGN), rectal temperature (TR), respiratory rate (FR), heart rate (FC), and body surface temperature (TSC), in the morning (7 AM) and in the afternoon (1 PM). The values of TR, TSC, RR and FR, mainly for the SS group, were higher in the afternoon. The TR, TSC and FR showed a significant and positive correlation with TA and the temperature and humidity index (ITGU), and a negative correlation with UR. These correlations were higher for the less rainy period. The FR showed significant and positive correlations with TA and ITGU, during both the most and the less rainy seasons, and negative correlations with UR only in the rainy season. The forestation of pasture areas is efficient to increase the thermal comfort for Murrah females, especially in the afternoon.

This study compared the two breed groups of Girolando (½ Holstein ½ Gyr vs. ¾ Holstein ¼ Gyr) through analysis of the percentages (stressed or non-stressed cows) of rectal temperature (RT), respiratory rate (RR) and pregnancy rate (PR), and means of production and reproduction parameters to determine the group best suited to rearing in semiarid tropical climate. The experiment was conducted at the farm, in the municipality of Umirim, State of Ceará, Brazil. Two hundred and forty cows were used in a 2 × 2 factorial study; 120 of each group were kept under an intensive system during wet and dry seasons. The environmental parameters obtained were relative humidity (RH), air temperature (AT), and the temperature and humidity index (THI). Pregnancy diagnosis (PD) was determined by ultrasonography 30 days after artificial insemination (AI). The milk production of each cow was recorded with automated milkings in the farm. The variables were expressed as mean and standard error, evaluated by ANOVA at 5% probability using the GLM procedure of SAS. Chi-square test at 5% probability was applied to data of RT, RR, pregnancy rate (PR), and the number of AIs to obtain pregnancy. The majority of ½ Holstein cows showed mean values of RT and RR within the normal range in both periods and shifts. Most animals of the ¾ Holstein group exhibited the RR means above normal during the afternoon in the rainy and dry periods and RT means above normal during the afternoon in the dry period. After analyses, ½ Holstein crossbred cows are more capable of thermoregulating than ¾ Holstein cows under conditions of thermal stress, and the dry period was more impacting for bovine physiology with significant changes in physiological parameters, even for the first breed group. Knowledge of breed groups adapted to climatic conditions of northeastern Brazil can directly assist cattle farmers in selecting animals best adapted for forming herds.

RESUMO -Um estudo foi conduzido com a finalidade de avaliar o efeito do estresse térmico sobre parâmetros seminais e biometria testicular em carneiros Santa Inês. Oito animais adultos foram submetidos à insulação escrotal por sete dias. Realizaram-se duas avaliações antes da insulação e, após este período, doze coletas durante 118 dias. A insulação escrotal não causou variações no volume dos ejaculados, mas o pH seminal apresentou valores elevados oito dias após a insulação, retornando à normalidade após 15 dias. Ocorreu redução na concentração espermática oito dias após a insulação e os animais apresentaram azoospermia entre 33 e 50 dias. Depois de 79 dias, a concentração espermática retornou aos padrões observados antes da insulação. A circunferência escrotal (26,4 cm) reduziu para 22,4 cm oito dias após o término do tratamento e para 21 cm aos 21 dias, mas retornou a valores próximos à normalidade (24,9 cm) aos 50 dias. Foi observado decréscimo na motilidade e vigor espermático a partir da retirada da bolsa e o retorno destes parâmetros à normalidade ocorreu somente após 90 dias. Antes da insulação, 1,7% dos espermatozóides apresentaram-se com defeitos maiores e 9,9 %, com defeitos menores. Na primeira coleta após a insulação, 3,6% das células apresentaram defeitos maiores e 43,4 %, defeitos menores; aos oito dias, estes valores foram de 8,4 e 60,4%, respectivamente. Entre 15 e 60 dias após a insulação, a percentagem de defeitos maiores variou entre 27,3 e 16,8%, enquanto a de defeitos menores permaneceu em torno de 39%. Aos 118 dias após a insulação, os defeitos maiores representaram somente 0,7%, mas o número de células com defeitos menores continuou elevado (24,4 %). Portanto, o estresse térmico causou interrupção temporária do processo de produção espermática e os parâmetros relativos à motilidade e defeitos menores apresentaram maior susceptibilidade.Palavras-chave: carneiros, estresse térmico, sêmen, testículos Effects of Scrotal Insulation on Testis Size and Semen Criteria in Santa Inês Hairy SheepRaised in the State of Ceará, Northeast of Brazil ABSTRACT -A study was conducted to determine the effect of heat stress on semen criteria and testis size in Santa Inês hairy rams, in the State of Ceará, Northeast of Brazil. The scrotum of eight adult rams was insulated for seven days. Animals were evaluated twice before insulation and, after insulation, at 12 different periods until 118 days. Scrotal insulation did not affect semen volume, but pH was increased after eight days, returning to normal values after 15 days. Sperm concentration was reduced eight days after insulation was removed and animals became azzospermic between 33 and 50 days. After 79 days, sperm concentration returned to values similar to those observed before treatment. Scrotal circumference (26.4 cm) decreased to 22.4 cm eight days after insulation and to 21 cm at 21 days, but returned to 24.9 cm after 50 days. Moreover, sperm motility and vigor were reduced right after insulation was removed and returned to normal profiles only after 90...

We have investigated the reproductive development of the tropically adapted Santa Inês ram, the most common hair sheep in Brazil. From 8 to 48 weeks of age, 16 animals were evaluated for body and testis growth, semen parameters, testosterone concentrations and seminal plasma proteins, using two-dimensional SDS-PAGE. Animals were weaned at 30 days and kept in feedlots thereafter, receiving hay, concentrate (18% of crude protein) and mineral supplement. Body weight increased from 12.3 +/- 0.7 to 54.3 +/- 1.6 kg between 8 and 48 weeks (p < 0.05), but changes in thoracic perimeter and scrotal circumference were non-significant after 36 weeks (p > 0.05). The percentage of motile sperm increased slowly until 23 weeks and more rapidly after that age, but significant changes in progressive motility occurred after 25 weeks. Presence of abnormal sperm related inversely to age. Most significant changes in sperm concentration occurred between 38 and 44 weeks (0.38 +/- 0.05 to 1.14 +/- 0.24 x 10(9) cells/ml, p < 0.05) and testosterone reached its highest concentrations at 42 weeks, decreasing afterwards. Rams reached puberty at 28.2 +/- 0.8 weeks. The number of protein spots on seminal plasma gels was similar from 15 to 18 weeks (45 and 47 spots; p > 0.05), increased until 24 weeks (141 spots) and 28 weeks (170 spots; p < 0.05) and remained without significant (p > 0.05) changes from 28 to 48 weeks (186 +/- 10 spots). Furthermore, the intensity of selected spots on 2D maps increased (p < 0.05) between 15 and 28 weeks, which preceded or coincided with the main developmental changes in sperm motility and percentage of defective sperm in the ejaculates. These results will support future studies designed to characterize specific seminal plasma proteins whose expression relate to the development of testis, epididymis and accessory sex glands.

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The study aimed to develop new and more specific thermal comfort indices for buffaloes reared in the Amazon region. Twenty female Murrah buffaloes were studied for a year. The animals were fed in pasture with drinking water and mineral supplementation ad libitum. The following parameters were measured twice a week in the morning (7 AM) and afternoon (1 PM): air temperature (AT), relative air humidity (RH), dew point temperature (DPT), wet bulb temperature (WBT), black globe temperature (BGT), rectal temperature (RT), respiratory rate (RR), and body surface temperature (BST). The temperature and humidity index (THI), globe temperature and humidity index (GTHI), Benezra's comfort index (BTCI), and Ibéria's heat tolerance index (IHTI) were calculated so they could be compared to the new indices. Multivariate regression analyses were carried out using the canonical correlation model, and all indices were correlated with the physiological and climatic variables. Three pairs of indices (general, effective, and practical) were determined comprising the buffalo comfort climatic condition index (BCCCI) and the buffalo environmental comfort index (BECI). The indices were validated and a great agreement was found among the BCCCIs (general, effective, and practical), with 98.3 % between general and effective a.nd 92.6 % between general and practical. A significant correlation (P < 0.01) was found between the new indices and the physiological and climatic variables, which indicated that these may be used in pairs to diagnose thermal stress in buffaloes reared in the Amazon.