Identical thermal stress coupled with different temperature and humidity combinations affects nutrient digestibility and gut metabolites of laying hens

The present study investigated whether the same temperature-humidity index (THI) values under different conditions of air temperature and relative humidity (RH) would affect the thermoregulatory, nutritional, and behavioral responses of laying hens. One hundred twenty Hy-Line Brown laying hens (60-weeks-old) were divided equally in two environmental chambers: 26 °C with 70% RH (hRH75) and 30 °C with 30% RH (hT75) for 28 days. The two ambient environments (hRH75 and hT75) had an identical THI value of 75, calculated using an empirical formula for laying hens. Neither hRH75 nor hT75 affected rectal and body-surface temperatures and heart and respiratory rates. The concentration of volatile fatty acids in fecal excreta were altered by the thermal treatments. hT75 vs. hRH75 decreased the proportion of acetate and increased the proportion of propionate in fecal samples. hT75 vs. hRH75 lowered the digestibility of dry matter, crude protein, and neutral detergent fiber at 14 days. Thermal treatments did not affect heat stress-associated behavioral responses including feeding, drinking, panting, and wing elevation at any stage. Laying hens exposed to the same THI at different temperatures and RH exhibit equal physiological responses including rectal and body-surface temperatures, heart and respiratory rates, and behavioral responses. Nonetheless, high-temperature treatment (hT75; 30 °C and 30% RH) vs. low temperature treatment (hRH75; 26 °C and 70% RH) affects nutrient digestibility and gut metabolites, suggesting that there are negligible but discernable responses to temperature in the gut physiology.(AU)

Seasonality effect on the physiological and productive responses of crossbred dairy cows to the equatorial Amazon climate / Efeito da sazonalidade sobre as respostas fisiológicas e produtivas de vacas leiteiras mestiças ao clima amazônico equatorial

The objective of this study was to evaluate the seasonal effect of months of the year upon the physiological and productive responses of crossbred dairy cows raised in an Amazonian climate. Twenty lactating cows were evaluated, fed on Brachiaria decumbens pasture, with free access to water and mineral supplementation. Data from climate variables air temperature (AT), relative humidity (RH), rainfall (RA) and temperature and humidity index (THI) were recorded dur ing the months of January to April 2019. The physiological data collected were: respiratory rate (RR, mov/min), heart rate (HR, beats/min), rectal temperature (RT, ºC), udder surface temperature (UST, ºC), body surface temperature (BST, ºC), dorsum surface temperature (DST, ºC), front surface temperature (FST, ºC) and rear shin temperature (RST). Milk production (MP) was also measured. There was a significant difference (P<0.05) of RST and RR with values ranging from 34.8 to 35.5°C and 32.0 to 36.2 mov/min, respectively. There were no significant difference (P>0.05) for BST, and the values for MP and THI were 3.8; 3.8; 4.6; 4.1 kg and 77.7; 79.7; 80.6; 80.1, respectively. It was concluded that there was a seasonal effect of the months of the year evaluated on the respiratory rate of animals, however, it did not change the MP and the THI. The Amazon environment is conducive to causing thermal stress in lactating cows raised on pasture, requiring the use of shading to facilitate the ability of these animals to dissipate heat.(AU)

O objetivo desse estudo foi avaliar os efeitos sazonais dos meses do ano sobre as respostas fisiológicas e produtivas de vacas leiteiras mestiças criadas em clima amazônico. Foram avaliadas 20 vacas lactantes, alimentadas com pasto de Brachiaria decumbens, com acesso livre a água e suplementação mineral. Foram registrados dados das variáveis climáticas temperatura do ar (TA), umidade relativa do ar (UR), precipitação pluviométrica (PP) e índice de temperatura e Umidade (ITU) durante os meses de janeiro a abril de 2019. Os dados fisiológicos coletados foram: frequência respiratória (FR, mov/min), frequência cardíaca (FC, bat./min), temperatura retal (TR, ºC), temperatura superficial do úbere (TSU, ºC), temperatura superfície corporal (TSC, ºC), temperatura superficial do dorso (TSD, ºC), temperatura superficial da fronte (TSF, ºC) e temperatura superficial da canela (TSCA, °C). Também foi mensurada a produção de leite (PL, kg). Houve diferença significativa (P<0,05) da TSCA e FR com valores variando de 34,8 a 35,5°C e 32,0 a 36,2 mov/min, respectivamente. Não houve diferença significativa (P>0,05) para TSC e os valores observados para PL e ITU foram 3,8; 3,8; 4,6; 4,1 kg e 77,7; 79,7; 80,6; 80,1, respectivamente. Houve efeito sazonal dos meses avaliados sobre a taxa respiratória dos animais, no entanto, isso não alterou a PL nem o ITU. O ambiente amazônico é propício a causar estresse térmico em vacas lactantes mantidas a pasto, sendo necessário o uso de sombreamento para facilitar a capacidade de dissipação de calor corporal desses animais.(AU)

Broiler behavior: influence of thermal stress, age, and period of the day

This study evaluated the behavior of broiler chickens subjected to thermal stress during different periods of the day and age of birds. A total of 180 one-day-old male broiler chicks were allocated to three chambers. Each chamber contained five boxes, with 12 chicks/box. Birds were assigned to a completely randomized design. The birds were filmed, and the images recorded every minute during the two periods: morning (from 08:00 to 09:00 h) and afternoon (from 17:00 to 18:00 h), at 7, 14, and 35 d of age. The frequency of the behavioral category (water intake, feed intake, resting, exploration, and comfort) was determined. Data were subjected to variance analysis for mixed models with rearing temperatures (cold, usual, and hot) and age of birds (7, 14, and 35 d) as subdivided plot, and periods of the day (morning and afternoon) as sub-subdivided plot. Birds at cold and usual temperatures showed higher feed intake than those at hot temperature in the morning. Birds exposed to cold and usual temperatures explored the environment less frequently at 35 d when compared with 14 d. However, chickens at hot temperature showed decreased exploration according to the increase of age. Birds spent more time feeding during the morning, regardless of age and rearing temperature. Also, during this period of the day, chickens spent most of the time exploring the environment. The frequency of feed intake and exploration decreases with the increase of age. The similarity of behavior between chickens reared at cold and usual temperatures may be due to a change in the thermal comfort zone of the birds. It suggests that the real thermal comfort temperature is between the two temperature ranges studied (cold and usual).

Tracking of thermal microbial shock proteins and genomic variation of Bacillus cereus due to some abiotic stresses / Rastreamento de proteínas de choque microbiano térmico e variação genômica de Bacillus cereus devido a alguns estresses abióticos

Bacillus cereus is considered the most potent bacterial strain in terms of the increment in induced proteins during thermal treatment at 52 °C for 90 min. Protein production in food-born microorganism (Bacillus cereus) recovered from contaminated food was investigated in response to heat shock treatment. Bacterial tolerance towards pH, salinity, and temperature at various levels was also investigated. Heat-shock proteins (HSPs) produced when exposed to 52 °C for up to 60 minutes led to significant differences (30%) above the untreated control (37 °C), and the maximum difference was recorded at 52°C at 90 minutes. ISSR detected a higher number of bands/primer than RAPD (13.7 vs. 12.7, respectively), and more polymorphic bands (10.7 vs. 8.4 bands/primer, respectively). The untreated bacterial strain did not grow at pH levels lower than 3, whereas the thermally treated strain grew significantly at pH two. A consistent increase in HSPs was observed, with a gradual increase in salinity of less than 16%. Surprisingly, the gradual increase in temperature did not induce tolerance against higher temperatures. However, a significant growth rate was noticed in response to heat-shocked treatments. The untreated Bacillus cereus demonstrated antibiotic resistance to gentamycin and clindamycin (1.54 and 1.65 cm, respectively), much lower than the corresponding inhibition areas with preheat-treated test pathogen which were recorded (2.37 and 2.49 cm, respectively).

Bacillus cereus é considerada a cepa bacteriana mais potente em termos de incremento de proteínas induzidas durante o tratamento térmico a 52 °C por 90 min. A produção de proteínas em microorganismos de origem alimentar (Bacillus cereus) recuperados de alimentos contaminados foi investigada em resposta ao tratamento de choque térmico. A tolerância bacteriana ao pH, salinidade e temperatura em vários níveis também foram investigadas. Proteínas de choque térmico (HSPs) produzidas quando expostas a 52 °C por até 60 minutos levaram a diferenças significativas (30%) acima do controle não tratado (37 °C), e a diferença máxima foi registrada a 52 °C em 90 minutos . O ISSR detectou um maior número de bandas/iniciador do que o RAPD (13,7 vs. 12,7, respectivamente) e mais bandas polimórficas (10,7 vs. 8,4 bandas/iniciador, respectivamente). A cepa bacteriana não tratada não cresceu em níveis de pH abaixo de 3, enquanto a cepa tratada termicamente cresceu significativamente em pH dois. Observou-se aumento consistente de HSPs, com aumento gradual da salinidade inferior a 16%. Surpreendentemente, o aumento gradual da temperatura não induziu tolerância a temperaturas mais altas. No entanto, uma taxa de crescimento significativa foi observada em resposta aos tratamentos de choque térmico. O Bacillus cereus não tratado demonstrou resistência antibiótica à gentamicina e clindamicina (1,54 e 1,65 cm, respectivamente), muito menor do que as áreas de inibição correspondentes com patógeno de teste pré-tratado que foram registradas (2,37 e 2,49 cm, respectivamente).

Evaluation of the thermoregulatory response of goats naturally exposed to the tropical climate using longitudinal data / Avaliação da resposta termorregulatória de cabras expostas naturalmente ao clima tropical a partir do uso de dados longitudinais

Rectal temperature (RT), heart rate (HR), and respiratory rate (RR), determined as repeated measurements over time in female goats, were used to identify covariance matrices that best fit the data for residual modeling on these three traits. Then, based on this result, the goats' responses to heat were evaluated. Five matrices were found with convergence for the three traits. The Heterogeneous Compound Symmetry matrix showed a good fit for modeling the residual associated with RT, whereas the Heterogeneous Autoregressive matrix had a better fit for RR and HR, according to the Akaike Information Criteria (AIC), corrected AIC (AICc), and Schwarz Bayesian Information Criterion (BIC) used. After adjusting the residual data for these three traits, a mixed-model analysis was used to evaluate collection period (3), physiological stage (3), and animal age (3) as fixed effects. Residual modeling interfered differently with the p-value associated with the fixed effects studied. Collection period and interactions did not influence the variation in RT (P>0.761), which was within the standard range for goats in the tropics, while the physiological stage of the goats affected it (P<0.05). Rectal temperature, HR, and RR tend to show covariance structures that can be modeled using specific residual covariance matrices, that is, the heterogeneous compound symmetry matrix best suits RT data, whereas the heterogeneous autoregressive matrix is better suited for HR and RR, which are usually correlated. The goats of the evaluated breed maintain RT within the range of variation displayed by breeds adapted to a hot environment, regardless of their physiological condition. Variations occur in RR and HR, without, however, exceeding the normal range for goats. Pregnancy causes goats to raise their RR in the rainy season of the year in the region in order to maintain RT within the normal range for the species.(AU)

Utilizou-se a Temperatura retal (TR), Frequências cardíaca (FC) e respiratória (FR) aferidas como medidas repetidas no tempo em fêmeas caprinas, objetivando-se identificar matrizes de estruturas de covariância que melhor se ajustou aos dados para modelagem do resíduo nessas três características e, em seguida, avaliou-se a respostas de cabras ao calor, com base nesse resultado. Constatou-se cinco matrizes com convergência nas três características. A Simétrica composta heterogênea ajustou-se bem para modelagem do resíduo associado a TR, enquanto a Autorregressiva heterogênea ajustou-se melhor para a FR e FC, de acordo com os critérios de informação de Akaike (AIC), Akaike corrigido (AICc) e o Bayesiano de Schwarz (BIC) utilizados. Com o resíduos de dados dessas três características ajustados, utilizou-se uma análise com modelos mistos para avaliar a Época de coleta (3), Estado fisiológico (3) e Idade do animal (3) foram como efeitos fixos. Constatou-se que a modelagem do resíduo interferiu de modo diferenciado no p valor associado aos efeitos fixos estudados. A época da coleta e interações não influenciaram a variação da TR (P>0,761), que oscilou dentro da faixa padrão para caprinos nos trópicos, mas o Estágio fisiológico da cabra sim (P<0,05). A Temperatura retal e as Frequências cardíaca e respiratória tendem a apresentar estruturas de covariâncias modeláveis com utilização de matrizes de covariâncias residuais especificas, ou seja, a matriz Simétrica composta heterogênea mais adequada para dados da Temperatura retal, enquanto a Autorregressiva heterogênea para as Frequências cardíaca e respiratória, geralmente correlacionas. As cabras da raça avaliadas mantêm a temperatura retal dentro da amplitude de variação apresentada por raças adaptadas a ambiente quente. Isso ocorre independente da condição fisiológica que se encontra, mas com ocorrência de variação na frequência respiratória e cardíaca, não excedendo, no entanto, a faixa normal para caprinos. A gestação condiciona a cabra a elevar a FR na época chuvosa do ano na região para manter a TR na faixa de amplitude normal para caprinos.(AU)

Thermo-resistant enzyme-producing microorganisms isolated from composting

Abstract Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

Resumo Os fertilizantes organo-minerais suplementados com aditivos biológicos são uma alternativa aos adubos químicos. Neste estudo, microrganismos termoresistentes foram isolados de compostagem por procedimentos de duas etapas. Inicialmente, as amostras tomadas em diferentes períodos e temperaturas (33 dias a 52 ºC, 60 dias a 63 ºC e mais de 365 dias a 26 ºC) foram pré-incubadas a 80 oC por 30 minutos. Posteriormente, a seleção microbiana foi conduzida por métodos baseados em cultura in vitro e choque térmico a 60 oC e 100 oC por 2h e 4h. Quarenta e um isolados foram capazes de crescer a 60 °C por 4h; vinte e sete a 100 °C por 2h e dois a 100 °C por 4h. A identificação molecular por sequenciamento parcial do gene ribossomal 16S usando primers universais revelou que trinta e cinco isolados eram de oito espécies de Bacillus, um Brevibacillus borstelensis, três Streptomyces thermogriseus e dois fungos (Thermomyces lanuginosus e T. dupontii). Os dados dos ensaios de atividade de amilase, fitase e celulase e o índice enzimático (IE) mostraram que 38 dos 41 isolados termorresistentes produziram pelo menos uma enzima. Para a atividade da amilase, o maior valor de IE foi observado em Bacillus licheniformis (isolado 21C2, IE = 4,11), seguido por Brevibacillus borstelensis (isolado 6C2, IE = 3,66), Bacillus cereus (isolado 18C2, IE = 3,52) e Bacillus paralicheniformis (isolado 20C2, IE = 3,34). Para a fitase, os maiores valores de IE foram observados para B. cereus (isolado 18C2, IE = 2,30) e B. licheniformis (isolado 3C1, IE = 2,15). Em relação à produção de celulose, B. altitudinis (isolado 6C1) foi o mais eficiente (IE = 6,40), seguido por três Bacillus subtilis (isolados 9C1, 16C2 e 19C2) com valores de IE de 5,66, 5,84 e 5,88, respectivamente, e um B. pumilus (isolado 27C2, IE = 5,78). Pode-se inferir que os microrganismos selecionados são potencialmente úteis como aditivos biológicos em fertilizantes organo-minerais e outros processos biotecnológicos.

Thermo-resistant enzyme-producing microorganisms isolated from composting / Microrganismos produtores de enzimas termo-resistentes isolados de compostagem

Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.

Os fertilizantes organo-minerais suplementados com aditivos biológicos são uma alternativa aos adubos químicos. Neste estudo, microrganismos termoresistentes foram isolados de compostagem por procedimentos de duas etapas. Inicialmente, as amostras tomadas em diferentes períodos e temperaturas (33 dias a 52 ºC, 60 dias a 63 ºC e mais de 365 dias a 26 ºC) foram pré-incubadas a 80 oC por 30 minutos. Posteriormente, a seleção microbiana foi conduzida por métodos baseados em cultura in vitro e choque térmico a 60 oC e 100 oC por 2h e 4h. Quarenta e um isolados foram capazes de crescer a 60 °C por 4h; vinte e sete a 100 °C por 2h e dois a 100 °C por 4h. A identificação molecular por sequenciamento parcial do gene ribossomal 16S usando primers universais revelou que trinta e cinco isolados eram de oito espécies de Bacillus, um Brevibacillus borstelensis, três Streptomyces thermogriseus e dois fungos (Thermomyces lanuginosus e T. dupontii). Os dados dos ensaios de atividade de amilase, fitase e celulase e o índice enzimático (IE) mostraram que 38 dos 41 isolados termorresistentes produziram pelo menos uma enzima. Para a atividade da amilase, o maior valor de IE foi observado em Bacillus licheniformis (isolado 21C2, IE = 4,11), seguido por Brevibacillus borstelensis (isolado 6C2, IE = 3,66), Bacillus cereus (isolado 18C2, IE = 3,52) e Bacillus paralicheniformis (isolado 20C2, IE = 3,34). Para a fitase, os maiores valores de IE foram observados para B. cereus (isolado 18C2, IE = 2,30) e B. licheniformis (isolado 3C1, IE = 2,15). Em relação à produção de celulose, B. altitudinis (isolado 6C1) foi o mais eficiente (IE = 6,40), seguido por três Bacillus subtilis (isolados 9C1, 16C2 e 19C2) com valores de IE de 5,66, 5,84 e 5,88, respectivamente, e um B. pumilus (isolado 27C2, IE = 5,78). Pode-se inferir que os microrganismos selecionados são potencialmente úteis como aditivos biológicos em fertilizantes organo-minerais e outros processos biotecnológicos.

Thermo-resistant enzyme-producing microorganisms isolated from composting / Microrganismos produtores de enzimas termo-resistentes isolados de compostagem

Organo-mineral fertilizers supplemented with biological additives are an alternative to chemical fertilizers. In this study, thermoresistant microorganisms from composting mass were isolated by two-step procedures. First, samples taken at different time points and temperatures (33 days at 52 ºC, 60 days at 63 ºC, and over 365 days at 26 ºC) were pre-incubated at 80 oC for 30 minutes. Second, the microbial selection by in vitro culture-based methods and heat shock at 60 oC and 100 oC for 2h and 4h. Forty-one isolates were able to grow at 60 °C for 4h; twenty-seven at 100 °C for 2h, and two at 100 °C for 4h. The molecular identification by partial sequencing of the 16S ribosomal gene using universal primers revealed that thirty-five isolates were from eight Bacillus species, one Brevibacillus borstelensis, three Streptomyces thermogriseus, and two fungi (Thermomyces lanuginosus and T. dupontii). Data from amylase, phytase, and cellulase activity assays and the enzymatic index (EI) showed that 38 of 41 thermo-resistant isolates produce at least one enzyme. For amylase activity, the highest EI value was observed in Bacillus licheniformis (isolate 21C2, EI= 4.11), followed by Brevibacillus borstelensis (isolate 6C2, EI= 3.66), Bacillus cereus (isolate 18C2, EI= 3.52), and Bacillus paralicheniformis (isolate 20C2, EI= 3.34). For phytase, the highest EI values were observed for Bacillus cereus (isolate 18C2, EI= 2.30) and Bacillus licheniformis (isolate 3C1, EI= 2.15). Concerning cellulose production, B. altitudinis (isolate 6C1) was the most efficient (EI= 6.40), followed by three Bacillus subtilis (isolates 9C1, 16C2, and 19C2) with EI values of 5.66, 5.84, and 5.88, respectively, and one B. pumilus (isolate 27C2, EI= 5.78). The selected microorganisms are potentially useful as a biological additive in organo-mineral fertilizers and other biotechnological processes.(AU)

Os fertilizantes organo-minerais suplementados com aditivos biológicos são uma alternativa aos adubos químicos. Neste estudo, microrganismos termoresistentes foram isolados de compostagem por procedimentos de duas etapas. Inicialmente, as amostras tomadas em diferentes períodos e temperaturas (33 dias a 52 ºC, 60 dias a 63 ºC e mais de 365 dias a 26 ºC) foram pré-incubadas a 80 oC por 30 minutos. Posteriormente, a seleção microbiana foi conduzida por métodos baseados em cultura in vitro e choque térmico a 60 oC e 100 oC por 2h e 4h. Quarenta e um isolados foram capazes de crescer a 60 °C por 4h; vinte e sete a 100 °C por 2h e dois a 100 °C por 4h. A identificação molecular por sequenciamento parcial do gene ribossomal 16S usando primers universais revelou que trinta e cinco isolados eram de oito espécies de Bacillus, um Brevibacillus borstelensis, três Streptomyces thermogriseus e dois fungos (Thermomyces lanuginosus e T. dupontii). Os dados dos ensaios de atividade de amilase, fitase e celulase e o índice enzimático (IE) mostraram que 38 dos 41 isolados termorresistentes produziram pelo menos uma enzima. Para a atividade da amilase, o maior valor de IE foi observado em Bacillus licheniformis (isolado 21C2, IE = 4,11), seguido por Brevibacillus borstelensis (isolado 6C2, IE = 3,66), Bacillus cereus (isolado 18C2, IE = 3,52) e Bacillus paralicheniformis (isolado 20C2, IE = 3,34). Para a fitase, os maiores valores de IE foram observados para B. cereus (isolado 18C2, IE = 2,30) e B. licheniformis (isolado 3C1, IE = 2,15). Em relação à produção de celulose, B. altitudinis (isolado 6C1) foi o mais eficiente (IE = 6,40), seguido por três Bacillus subtilis (isolados 9C1, 16C2 e 19C2) com valores de IE de 5,66, 5,84 e 5,88, respectivamente, e um B. pumilus (isolado 27C2, IE = 5,78). Pode-se inferir que os microrganismos selecionados são potencialmente úteis como aditivos biológicos em fertilizantes organo-minerais e outros processos biotecnológicos.(AU)

Molecular investigations of the effect of thermal manipulation during embryogenesis on muscle heat shock protein 70 and thermotolerance in broiler chickens

The objective of this study was to elucidate the optimum protocol timing of thermal manipulation (TM) during embryogenesis, which underline genetic improvement of muscle thermotolerance acquisition. For the present study, 1,440 fertile eggs were divided randomly and equally into control (37.8 °C with 56% relative humidity) and four thermally manipulated groups (TM1, TM2, TM3, and TM4) subjected to 39 °C for 18 h with 65% relative humidity daily during different embryonic periods. Then, at day 35 post-hatch, all groups were subjected to thermal challenge at 43 °C for 6 h to identify the level of thermotolerance acquisition differences between them. Hsp70 mRNA expression was evaluated by using a relative quantitatively RT-qPCR. Single nucleotide polymorphisms sequence of the Hsp70 gene was evaluated by Sanger's sequencing method. Pectoral and thigh muscles samples were subjected to immunohistochemistry to detect Hsp70. Among TM conditions that were investigated, TM1 (39 °C for 18 h during embryonic days (ED) 7­11) induced a significant improvement in thermotolerance parameters (body temperature and T3 levels) during thermal challenge combined with an increase in the levels of Hsp70 mRNA and its protein with a high stability of nucleotide sequences in both pectoral and thigh muscles. The partial DNA sequence of Hsp70 gene in TM1 was reported, and nucleotide sequences were deposited in NCBI GenBank database with the accession numbers (MK852579) and (MK852580). Thigh muscle thermotolerance acquisition was higher than pectoral muscle during thermal challenge at 43 °C for 6 h. Thus, TM during ED7­11 may improve thermotolerance acquisition without adversely affecting performance.(AU)

Magnesium in the water of Japanese quails kept under comfort zone and under thermal stress / Magnésio na água de codornas japonesas mantidas em zona de conforto e sob estresse térmico

The objective was to evaluate the increasing levels of magnesium in the water supplied to laying quails (Coturnix coturnix japonica), kept in climatic chambers under thermoneutral temperature and thermal stress, on their performance and morphometry of their organs. The birds were distributed in a completely randomized design, 2x4 factorial arrangement, 2 temperatures (24 and 32 ºC) and 4 levels of magnesium in the water (50, 150, 250 and 350 mg L-1), with six replicates and eight birds per experimental unit. The data were subjected to analysis of variance and the means compared by the Tukey test at 5% probability level. The magnesium levels in the water did not affect (P > 0.05) the production performance and morphometry of the organs, with less water consumption at the magnesium level of 150 mg L-1, and birds kept at 32 °C had a reduction in feed consumption and feed conversion, but without affecting organ morphometry. Japanese quails in the production phase can consume water with magnesium levels up to 350 mg L-1 without having their production performance and morphometry of organs affected and raised in an environment with temperatures of up to 32 °C.(AU)

O objetivo foi avaliar os níveis crescentes de magnésio na água fornecida a codornas (Coturnix coturnix japonica) em postura, mantidas em câmaras climáticas sob temperatura termoneutra e estresse térmico, sobre o desempenho e morfometria dos órgãos das aves. As aves foram distribuídas em delineamento inteiramente casualizado, arranjo fatorial 2x4, 2 temperaturas (24 e 32 ºC) e 4 níveis de magnésio na água (50, 150, 250 e 350 mg L-1), com seis repetições e oito aves por unidade experimental e os dados submetidos à análise de variância e as médias comparadas pelo teste de Tukey a 5% de probabilidade. Os níveis de magnésio na água não afetaram (P > 0,05) o desempenho produtivo e a morfometria dos órgãos, com menor consumo de água ao nível de 150 mg L-1 de magnésio e, as aves mantidas a 32 °C tiveram uma redução no consumo de ração e conversão alimentar, mas sem afetar a morfometria dos órgãos. Codornas japonesas em fase de produção podem consumir água com níveis de magnésio de até 350 mg L-1 sem ter o desempenho produtivo e a morfometria dos órgãos afetados e criados em ambiente com temperatura de até 32 °C.(AU)

Induction to tetraploidy in Pacific oysters (Crassostrea gigas)

As an alternative to the use of cytochalasin B(CB), 6-dimethylamino-purine (6-DMAP) and thermal shock (heat shock by increasing the temperature from 25 to 36ºC) could be used to induce tetraploidy in Pacific oyster (Crassostrea gigas) diploids. Induction was performed by applying shocks after elimination of the first polar corpuscle at the end of meiosis I. Ploidy rates were verified using flow cytometry. Tetraploid larvae were obtained using all inductor (6-DMAP, thermal shock and CB) treatments. No difference in the efficiency of tetraploidy induction was noted among 6-DMAP, thermal shock and CB. The number of D-larvae and their yield, determined by calculating the percentage of well-formed D-larvae in relation to the total number of larvae, was similar (p>0.05) among the evaluated induction methods. We suggest that 6-DMAP and thermal shock should be used in tetraploidy induction protocols, thereby avoiding the use of CB, which is a harmful agent for both humans and the environment.(AU)

Interactive effects of digestible protein levels on thermal and physical stress responses in Nile tilapia

The effects of dietary digestible protein (DP) levels (22, 26, 29, 32, and 34%) and different stressors (cold-induced stress, CIS; heat/dissolved oxygen-induced stress, HDOIS; transport-induced stress, TIS; and size-sorting-induced stress, SSIS) on hemato-biochemical parameters were evaluated. Four hundred and forty Nile tilapia fingerlings were distributed into 40-250 L aquaria and fed experimental diets for 110 days, and fed each of the five experimental diets, that were randomly distributed to eight replicates per treatment. Then, different groups of fish were subjected to one type of stress. Groups of 40 fish were used on CIS (17 °C), HDOIS (32 °C), and TIS (4 h), and a group of 140 fish on SSIS (15 min air exposure and 60 s handling). There was no effect on hemato-biochemical profile when DP levels were compared, neither before nor after stress; however, there was a significant stress effect. Digestible protein did not mitigate stress response under SSIS and CIS; lymphopenia and neutrophilia were the main cell-mediated immune response; dietary 22 and 26% DP impaired oxygenation on SSIS and TIS; fish under HDOIS and SSIS demanded more energy using triglycerides as an energy source; the diet formulated to contain 22% DP was not adequate to keep homeostasis under temperature stress. Cluster analysis showed that, for DP levels below the requirement for growth, SSIS and CIS were considered the most stressful conditions. At 34% DP level, HDOIS response was comparable to that of non-stressing conditions.(AU)

Effect of In Ovo Ascorbic Acid Injection and Thermal Manipulation During Incubation on Intestine Morphology of Broilers Reared under Cold and Heat Stress

It was investigated if pre-incubation ascorbic acid (AA) injection in fertile eggs incubated at high temperature impacts the performance, the yield of carcass and parts, and the intestine morphology of broilers reared under heat stress. Three thousand Cobb® fertile broiler eggs were randomly distributed according to weight into three incubations treatments (eggs not injected with AA and incubated at 37.5°C; eggs not injected with AA and incubated at 39°C; and eggs injected with 6 µg AA/100 µL water prior to incubation and incubated at 39ºC). The hatched birds were reared at thermoneutral, cold, and hot house temperatures. Broilers reared under hot temperature presented lower feed intake and weight gain than the broilers of the different rearing temperatures. Egg incubation at 39.0 ºC and 39.0 ºC + AA reduced broiler viability. Carcass and cut yields were not influenced by incubation and rearing procedures. Duodenal goblet cell count was lower in broilers from eggs of the treatment 39ºC + AA than in broilers from the other incubation treatments and in broiler rearing in hot temperature. In the jejunum, the goblet cell counts were higher in broilers that were reared under hot than thermoneutral temperatures. The incubation treatment of 39 ºC+AA increased the goblet cell counts in the ileum of broilers reared under cold temperatures. Rearing temperature influenced the duodenal villi counts, which were lower under cold rearing conditions than in the two other rearing temperatures. The results showed that egg incubation at 39°C, independently of ascorbic acid injection, did not produce an effective epigenetic heat adaptation in broilers.(AU)

Foraging Oxaea flavescens bees as a function of the dynamics of abiotic factors and food resource availability from Styrax camporum flowers

We examined the abiotic factors and co-specific competition for food resources that regulate the foraging activity of Oxaea flavescens bees on Styrax camporum flowers. Foraging records were gathered during 20 min. periods at the beginning of each hour between 05:00h and 18:00h during three nonconsecutive days. Pearson correlation and linear regression tests indicated that the foraging activity of O. flavescens was associated with abiotic factors during the day. O. flavescens represented 89.9% of the observed foraging visits to S. camporum flowers. On the first day of sampling, when environmental conditions were stressful, the foraging activity of O. flavescens was significantly negatively correlated with light intensity, wind speed, and temperature, and positively correlated with relative humidity. Under those conditions, optimal foraging was little affected by the availability of floral resources. On the second and third days, however, when environmental conditions were more favorable, the principal limiting factor of O. flavescens foraging activity was nectar depletion. The maximum peak of foraging under those conditions occurred before the abiotic conditions were fully favorable, however, as the eventual depletion of floral resources resulted in unfavorable cost/benefit implications for foraging during the otherwise most adequate daylight period.(AU)

Efeitos da tosquia no conforto térmico e desempenho produtivo de coelhos (Oryctolagus cuniculus f. domesticus) / Effects of shearing on thermal comfort and productive performance of rabbits (Oryctolagus cuniculus f. domesticus) / Efectos de la esquila sobre el confort térmico y el rendimiento productivo de conejos (Oryctolagus cuniculus f. domesticus

O coelho é uma das espécies mais sensíveis às condições do meio que vivem, sendo a temperatura e umidade os fatores que mais têm influência no seu ambiente. O objetivo com este trabalho foi avaliar a tosquia como alternativa para melhorar o conforto térmico e o desempenho produtivo de coelhos criados de forma intensiva em uma região de clima semiárido. Foram utilizados 26 coelhos mestiços, sadios e em fase de engorda, sendo divididos em gaiolas individuais em dois grupos: treze animais submetidos a tosquia (G1) e treze sem tosquia (G2). O conforto térmico foi avaliado todos os dias pelo cálculo de Índice de Temperatura e Umidade (ITH) e pela análise e comparação dos parâmetros: frequência respiratória (FR), temperatura retal (TR) e temperaturasuperficial (TS). A temperatura e umidade do ambiente foram verificadas em todos os dias do experimento, às 6:00, 9:00, 12:00, 15:00 e 18:00 horas, com um termo-higrômetro. O desempenho dos animais foi avaliado pela pesagem nos dias 0, 10, 20 e 30 do experimento. Como resultado, a FR e TS apresentaram diferença estatística (P<0,05), o grupo dos animais tosquiados apresentou média inferior de FR (221,6 ± 15,4) em relação aos não tosquiados (240,2 ± 26,1). Em relação à TS, houve menores temperaturas nos animais tosquiados (33,8°C ± 0,3) quando comparados com os não tosquiados (35,0°C ± 0,2). Quanto à TR, ficou próximo de 39,5 ºC, inferindo assim que os mecanismos de regulação térmica interno dos animais conseguiram manter a homeotermia desejada. O ganho de peso obteve média de 771,4 gramas no grupo dos animais tosquiados, e 705,2 gramas no grupo dos não tosquiados. Em relação à TR e desempenho dos animais, não foram observadas diferenças estatísticas (P > 0,05). De acordo com os dados obtidos, conclui-se que atosquia pode ser um meio de melhorar o conforto térmico de coelhos.(AU)

The rabbit is one of the most sensitive species to the conditions of the environment in which they live, with temperature and humidity being the factors that most influence their environment. The objective of this work was to evaluate shearing as an alternative to improve the thermal comfort and productive performance of rabbits raised intensively in a region with a semi-arid climate. Twenty-six mixed, healthy and fattening stage rabbits were used and divided into individual cages into two groups: thirteen animals submitted to shearing (G1) and thirteen without shearing (G2). Thermal comfort was evaluated every day by calculating the Temperature and Humidity Index (THI) and by analyzing and comparing the parameters: respiratory rate (RR), rectal temperature (RT) and superficial temperature (ST). The temperature and humidity of the environment were checked every day of the experiment, at 6:00, 9:00, 12:00, 15:00 and 18:00 hours, with a thermo-hygrometer. The performance of the animals was evaluated by weighingon 0, 10, 20 and 30 days of the experiment. As a result, the RR and ST showed a statistical difference (P<0.05), the group of sheared animals had a lower mean of RR (221.6 ± 15.4) in relation to the no sheared animals (240.2 ± 26.1). Regarding the ST, there were lower temperatures in the sheared animals (33.8°C ± 0.3) when compared to the no sheared animals (35.0°C ± 0.2). As for the RT was close to 39.5 ºC, inferring that the thermal regulation mechanisms of the animals managed to maintain the desired homeothermia. Weight gain averaged 771.4 grams in the sheared group, and 705.2 grams in the no sheared group. Regarding the RT and performance of the animals, no statistical differences were observed (P>0.05). According to the data obtained, it is concluded that shearing can improve the thermal comfort of rabbits.(AU)

El conejo es una de las especies más sensibles a las condiciones del medio en el que vive, siendo la temperatura y la humedad los factores que más influyen en su entorno. El objetivo de este trabajo fue evaluar la esquila como alternativapara mejorar el confort térmico y el desempeño productivo de conejos criados de forma intensiva en una región de clima semiárido. Se utilizaron 26 conejos mestizos, sanos y en fase de engorde, divididos en jaulas individuales en dos grupos: trece animalessometidos a esquila (G1) y trece sin esquila (G2). El confort térmico se evaluó diariamente mediante el cálculo del Índice de Temperatura y Humedad (ITH) y mediante el análisis y comparación de los parámetros: frecuencia respiratória (FR), temperatura rectal (TR) y temperatura superficial (TS). La temperatura y humedad del ambiente se verificado todos los días del experimento, a las 6:00, 9:00, 12:00, 15:00 y 18:00 horas, con un termohigrómetro. El rendimiento de los animales se evaluó mediante pesaje los días 0, 10, 20 y 30 del experimento. Como resultado, el FR y el TS mostraron diferencia estadística (P<0,05), el grupo de animales esquilados tuvo una media de FR más baja (221,6 ± 15,4) en relación a los animales no esquilados (240,2 ± 26,1). En cuanto a la TS, hubo temperaturas más bajas en los animales esquilados (33,8 °C ± 0,3) en comparación con los animales no esquilados (35,0 °C ± 0,2). En cuanto a la TR, fue cercana a los 39,5 ºC, infiriendo que los mecanismos internos de regulación térmica de los animales lograron mantener la homeotermia deseada. El aumento de peso promedió 771,4 gramos en el grupo esquilado y 705,2 gramos en el grupo no esquilado. En cuanto al TR y rendimiento de los animales, no se observaron diferencias estadísticas (P > 0,05). De acuerdo con los datos obtenidos, se concluye que las esquila puede ser un medio para mejorar el confort térmico de los conejos.(AU)

Incubation parameters, offspring growth, and behavioral adaptations to heat stress of Black Skimmers (Rynchops niger) in a Neotropical inland colony (Aves, Charadriiformes, Laridae)

This study focuses on incubation parameters, egg morphometrics, and body mass development, hatching, and behavioral adaptations to heat stress within a colony of freshwater-breeding Black Skimmers (Rynchops niger) located in the private nature reserve of Serviço Social do Comércio (SESC) in the northern Pantanal, Mato Grosso, Brazil. Temperatures of nest, eggs, and surface substratum, as well as the development of embryos, were surveyed using thermal imaging, a method allowing digital recording from a distance and in a fraction of the time of traditional measuring techniques. The mean egg dimensions (n = 71) were 4.48 (± 0.13) × 3.27 (± 0.07) cm; the mean mass at hatching was 24.3 (± 1.9) g, with a significant decrease over incubation time. The mean surface temperature of eggs varied from 30.9℃ to 39.7℃, while the sand surface temperature was 20℃ at 06:00 h, rising to 47.7℃ at 11:00 h. There was a significant increase (7%) in egg surface temperature throughout incubation. Incubation-bout durations (n = 2108) were correlated with the microclimatic conditions of the substratum, becoming shorter with increasing sand-surface temperature around midday. Egg hatching lasted one day, and siblings hatched no more than 24 h apart. The mean body mass on Day 1 after hatching was 16.8 (± 1.6) g (n = 6). Three days after hatching, chicks moved to new sand depressions provided by parents near the original nest, where they remained motionless or tried to hide under riparian vegetation. The single chick that fledged had a growth rate of K = 0.117 and a t10₋90 value of 37.3 days. On Day 7, dorsal pintail feathers and primaries appeared, which were open on Day 15. After 14 days, the chick was able to regulate its body temperature, and no more feeding by parental birds during the daytime was observed. On Day 21, the immature plumage was fully developed. Fledging was completed on Day 27. Our study demonstrates that thermal imaging is a useful method of surveying egg and embryo development in the Black Skimmer, reducing nest disturbance and observation efforts.(AU)

Performance and carcass and meat quality of crossbred sheep confined with and without access to artificial shade / Desempenho e qualidade de carcaça e carne de ovinos mestiços confinados com e sem acesso ao sombreamento artificial

The aim of this study was to evaluate the performance and carcass and meat quality of sheep in an intensive production system with and without artificial shade available. Twenty Santa Inês × Dorper crossbred lambs were allotted to two collective stalls, one of which was covered with a black polypropylene shade net with 80% light interception, providing 2.5 m2 shade animal-1. Performance was evaluated by measuring initial weight, final weight, carcass yield, average daily weight gain, and total weight gain. Carcass and meat traits were evaluated based on the variables of fatness, conformation, loin-eye area, subcutaneous fat thickness, marbling, texture, pH, temperature, and color. Performance and carcass and meat traits were similar between the animals reared in stalls with and without shade available. The groups with and without access to shade showed initial weights of 26.52 and 25.37 kg, final weights of 35.59 and 34.76 kg, carcass yields of 45.73 and 45.50%, loin-eye areas of 14 .11 and 13.61 cm2, and marbling scores of 4.78 and 4.44, respectively. Subcutaneous fat thickness (2.02 cm), texture (3.67), and fatness (2) values were equal for both groups. Based on the results, artificial shading did not affect the performance or carcass and meat quality of the sheep.(AU)

Objetivou-se avaliar o desempenho, qualidade de carcaça e carne de ovinos em sistema intensivo de produção, com e sem disponibilidade de sombra artificial. Foram utilizados 20 cordeiros mestiços Santa Inês x Dorper, distribuídos em duas baias coletivas, onde em uma das baias foi implantado o sombrite de malha de polipropileno, na coloração preta, com 80% de interceptação luminosa, respeitando 2,5m2 de sombra. animal-1. No desempenho foi avaliado o peso inicial, peso final, rendimento de carcaça, ganho de peso médio diário e ganho de peso total. Para as características de carcaça e carne, foram observados o acabamento, conformação, área de olho de lombo, espessura de gordura subcutânea, marmoreio, textura, pH, temperatura e coloração. Foram observados valores semelhantes de desempenho e características de carcaça e carne entre os animais criados em baias com e sem acesso ao sombreamento. O peso inicial foi de 26,52 kg e 25,37 kg, peso final de 35,59 kg e 34,76 kg, com rendimento de carcaça de 45,73% e 45,50%, área de olho de lombo de 14,11 cm2 e 13,61 cm2, marmoreio de 4,78 e 4,44 para os animais com e sem sombreamento, respectivamente. Os valores de espessura de gordura subcutânea (2,02 cm), textura (3,67) e acabamento (2) foram iguais para ambos os grupos de animais. Pelos resultados obtidos, considera-se que o sombreamento artificial não interferiu no desempenho e qualidade de carcaça e carne ovina.(AU)

Evaluation of environmental parameters in a microregion in southern Amazonas State, Brazil, and their relationship with heat stress in dairy cattle / Avaliação de parâmetros ambientais em uma microrregião no sul do estado do Amazonas e suas relações com estresse térmico de bovinos leiteiros

Environmental thermal comfort provides higher efficiency in the exploitation of dairy farming, as the responses of animals to the environment are decisive in the activity's success. This study aimed to evaluate heat stress in dairy cows in the municipality of Humaitá, located in the south of theAmazonas State (AM), Brazil.Air temperature (TA) and relative humidity (RH) data were used to calculate the temperature humidity index (THI) in the dry (August 2020) and rainy seasons (January 2021). The August data were collected at the automatic weather station belonging to the National Institute of Meteorology. The January data were collected by a Hobo® U23-001 thermo-hygrometer installed at Fazenda Nossa Senhora Aparecida. The hourly and monthly means of the data were calculated and, subsequently, boxplots were plotted. TA and RH were 0.65 °C and 0.64% below the provisional climatological normal (PCN), respectively, for August, while January presented 0.77 °C and 3.61% above the PCN, respectively. THI values ranged from 56.25 (comfortable) to 84.68 (emergency) in August and 74.15 (comfortable) to 84.07 (emergency) in January. The maximum THI was observed in August, as well as the largest range. The results allow inferring that heat stress in dairy cows in the municipality of Humaitá-AM presents a state of alert even for cross-breed animals. Thus, the use of shading in pastures is suggested to minimize the effect of heat stress on the productive performance of the animals.(AU)

O conforto térmico ambiental proporciona uma maior eficiência na exploração da pecuária leiteira, pois as respostas do animal ao ambiente são determinantes no sucesso da atividade. O objetivo deste trabalho foi avaliar o estresse térmico em vacas leiteiras no município de Humaitá, localizado no sul do estado do Amazonas (AM), Brasil. Foram utilizados dados de temperatura do ar (TA) e umidade relativa do ar (UR) para cálculo do Índice de Temperatura e Umidade (ITU) na estação seca (agosto de 2020) e na estação chuvosa (janeiro de 2021). Para o mês de agosto os dados foram coletados na estação meteorológica automática pertencente ao Instituto Nacional de Meteorologia. Em janeiro os dados foram coletados com o Termo-higrômetro (Hobo®, modelo U23-001) instalado na Fazenda Nossa Senhora Aparecida. As médias horárias e mensais dos dados foram feitas e, posteriormente, plotados gráficos boxplots. Os dados coletados para os meses do estudo apresentaram a TAe UR para o mês de agosto 0,65ºC e 0,64% abaixo da Normal Climatológica Provisória (NCP), respectivamente. Por outro lado, janeiro apresentou 0,77ºC e 3,61% acima da NCP, respectivamente. Os ITUs variaram entre 56,25 (confortável) e 84,68 (emergência), em agosto e de 74,15 (confortável) a 84,07 (emergência) em janeiro. O máximo ITU foi observado em agosto, bem como a maior amplitude. Baseados nos resultados encontrados pode-se aferir que o estresse térmico em vacas leiteiras no município de Humaitá-AM apresenta estado de alerta, mesmo para animais de raças cruzadas. Assim, sugere-se o uso de sombreamento nas pastagens para minimizar o efeito do estresse térmico no desempenho produtivo destes animais.(AU)

Infuence of heat stress on in vitro oocyte and embryo production in high-yielding Holstein cows / Influência do estresse calórico na produção in vitro de oócitos e embriões de vacas Holandesas de alta produtividade

The objective of this study was to evaluate the influence of thermal shock on oocytes used in the production of in vitro embryos (IVP) of high productivity Holstein cows on the day of follicular aspiration (OPU; 0), 30, 60 and 90 days before the OPU. From the mean temperature on day 0 and on the previous 30, 60 and 90 days, they were classifed into comfort group (TC; up to 15°C) and heat stress (HS; above 15°C) groups.Anegative influence was observed on oocytes and viable embryos (total and grade I). The heat stress in the periods of 30 and 60 days prior to OPU resulted in lower production of viable oocytes (P=0.0028; P=0.0092, respectively). Under stress, on the day of OPU (HS-OPU), cows showed no reduction in the amount of viable oocytes (P=0.5497) and there was no influence of temperature for the group stressed 90 days before OPU (P=0.8287). For total embryos, the difference occurred only in the HS-30 group (P=0.0317), where the groups HS-OPU, HS-60, HS-90 presented, respectively, P=0. 1987, P=0.0596 and P=0.4580. Regarding the production of embryos of grade 1, there was no difference for the groups HS-OPU (P=0.2291) and HS-90 (P=0.2868), but there was a reduction for HS-30 (P=0.0143) and HS-60 (P=0.0253). In summary, heat stress had a negative impact when it occurred 30 or 60 days before follicular aspiration. In addition, 30 days seems to be the period of more susceptibility and that causes the greatest deleterious effects on oocyte viability and IVP.(AU)

Objetivou-se avaliar a infuência do estresse térmico em oócitos utilizados na produção in vitro de embriões (PIV) bovinos da raça Holandesa de alta produtividade no dia da aspiração folicular (OPU; 0), 30, 60 e 90 dias antes da OPU. A partir da temperatura média no dia 0 e aos 30, 60 e 90 dias anteriores, foram classificados nos grupos conforto (CT; até 15°C) e estresse por calor (ET -acima de 15°C). Observou-se infuência negativa em oócitos e embriões viáveis (total e grau I). A submissão ao estresse térmico nos períodos de 30 e 60 dias anteriores à OPU resultou em menor produção de oócitos viáveis (P=0,0028; P=0,0092, respectivamente). Sob estresse, no dia da OPU (ET-OPU), as vacas não apresentaram redução na quantidade de oócitos viáveis (P=0,5497) e não houve infuência da temperatura para o grupo estressado 90 dias antes da OPU (P=0,8287). Para embriões totais, a diferença ocorreu apenas no grupo ET-30 (P=0,0317), onde os grupos ET-OPU, ET-60, ET-90 apresentaram, respectivamente, P=0,1987, P=0,0596 e P=0,4580. Em relação à produção de embriões grau 1, não houve diferença para os grupos ET-OPU (P=0,2291) e ET-90 (P=0,2868), porém houve redução para ET-30 (P=0,0143) e ET- 60 (P=0,0253). Em resumo, o estresse por calor teve impacto negativo quando ocorreu 30 ou 60 dias antes da aspiração folicular. Além disso, 30 dias parece ser o período de maior suscetibilidade e que causa os maiores efeitos deletérios na viabilidade oocitária e na PIV.(AU)

Sperm cell capacitation status of ram semen after cooling

Background: The use of conventional artificial insemination (AI) in sheep production is usually associated with lower fertility rates when frozen semen is used. Cooled ram semen has been an alternative over frozen semen due to the higher viability, seminal quality and fertility rates following AI. The semen preservation process promotes sperm cell modifications similar to capacitation (capacitation-like) that causes cell damage affecting viability and seminal quality, but such effects are unclear for cooled semen. The aim of this study was to determine the status of sperm cell capacitation (CA) and acrosome reaction (AR) during ram semen processing and cooling under different extenders, dilution factors, and aerobiosis conditions as a function of storage time at 5o C. Materials, Methods & Results: Two consecutive ejaculates per day per male were collected from 2 adult rams by artificial vagina at 48-72 h intervals, in three replications. After macro- and microscopic evaluations, semen was segregated into groups under 3 extenders (Tris-egg yolk or TY, citrate-egg yolk or CY, skimmed milk or SM), 2 dilution factors (1 x 109 or Bi, 100 x 106 or Mi cells/mL), and 2 aerobiosis conditions (aerobic or A, semi-anaerobic or SA). Diluted semen was cooled to 5ºC and stored for up to 72 h, with evaluations every 24 h. Aliquots of fresh ejaculates and of each cooled diluted subgroup, according to extender, dilution, and aerobiosis, were collected at times T0 and T72 for determination of acrosome status and membrane integrity by the chlortetracycline (CTC) and trypan blue-Giemsa stainings, respectively. No differences were detected in sperm cell motility (M) and motility vigor (V) between fresh and diluted semen. After cooling, a significant decrease in M was observed after 48 h in CY and SM compared with fresh semen and 0 h of cooling, while V started to decrease after 24 h in CY compared with TY. Likewise, M/V from different dilutions and aerobic conditions decreased more significantly after 48 and 24 h of cooling, respectively. The sperm capacitation status did not show differences in the proportion of non-capacitated (NCA), CA and AR sperm cells between TY, CY, and SM extenders (NCA: 75.0%, 71.3%, 74.0%; CA: 15.7%, 17.2%, 15.9%; AR: 9.3%, 11.5%, 10.2%) or between Bi and Mi dilutions (NCA: 74.0%, 72.9%; CA: 15.9%, 16.6%; AR: 10.1%, 10.5%), respectively. However, differences (P < 0.05) were observed between A and SA aerobic conditions, with CA (17.0% vs. 15.5%) and AR (11.9% vs. 8.7%) rates being higher in A than SA, respectively, with no differences in NCA (71.1% vs. 75.8%), irrespective of the storage time. Sperm cell viability decreased after 48 h, especially in CY (P < 0.05). Discussion: Ram sperm cells can suffer irreversible damage due to thermal shock during cooling. Egg yolk-based extenders provide phospholipids and cholesterol to protect the sperm cell membrane during the thermal shock caused by the change in temperature. In this study, sperm cells had irreversible decreases in M/V, with increase in acrosome and plasma membrane damage after cooling to 5ºC. The largest and smallest decreases in M and V over time were observed in the CY and TY extenders, respectively. In addition to the extender type, the semen preservation method and storage time promoted changes in the capacitation status, AR and in sperm cell viability, which per se were associated with a decrease in semen fertility. In fact, the proportions of CA and/or AR sperm cells gradually increased over time after dilution and storage at 5ºC, with a negative correlation between sperm cell viability and M/V over time. In summary, extender and cooling time affected mostly M/V, while aerobiosis condition and dilution factor were more associated with acrosome status and sperm survival, with the extender having less impact on the acrosome status as a function of time.