The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells.

The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-old's lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V-FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days' culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.

The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells

The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-olds lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V–FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.(AU)

Nano-selenium and nano-zinc oxide supplementation in syrup on laying area, population size and hsp gene expression of honey bees in hot climate

This study was designed to examine the protective effects of nano-selenium and nano-zinc oxide on queen and workers performance under heat stress condition and gene expression of heat shock protein 70 (hsp70) as an index of heat tolerance. Sixty colonies were randomly assigned to five treatments with 12 replicates from June until early September. Sugar syrup (50%) containing no supplement or nano-selenium at levels of 50 and 100 µg L-1 or nano-zinc at levels of 100 and 200 µg L-1 was fed to colonies. Nano-selenium supplementations had no effect, but nano-zinc at level of 100 µg L-1 significantly decreased body malondialdehyde concentration. The highest bee population was seen in nano-zinc at level of 100 µg L-1 and the lowest one in control group. The lowest and the highest body weight, fat and protein deposition was found in group received nano-zinc at level of 100 µg L-1 and control, respectively. The highest gene expression was for group received nano-zinc at level of 100 µg L-1 In group received nano-zinc at level of 100 µg L-1, an increase in hsp70 gene expression was found. In conclusion, nano-zinc oxide at level of 100 µg L-1 could increase queen and worker performance and heat resistance of bees in the hot climate condition.(AU)

The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells

Abstract The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-old's lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V-FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days' culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.

Nano-selenium and nano-zinc oxide supplementation in syrup on laying area, population size and hsp gene expression of honey bees in hot climate

This study was designed to examine the protective effects of nano-selenium and nano-zinc oxide on queen and workers performance under heat stress condition and gene expression of heat shock protein 70 (hsp70) as an index of heat tolerance. Sixty colonies were randomly assigned to five treatments with 12 replicates from June until early September. Sugar syrup (50%) containing no supplement or nano-selenium at levels of 50 and 100 µg L-1 or nano-zinc at levels of 100 and 200 µg L-1 was fed to colonies. Nano-selenium supplementations had no effect, but nano-zinc at level of 100 µg L-1 significantly decreased body malondialdehyde concentration. The highest bee population was seen in nano-zinc at level of 100 µg L-1 and the lowest one in control group. The lowest and the highest body weight, fat and protein deposition was found in group received nano-zinc at level of 100 µg L-1 and control, respectively. The highest gene expression was for group received nano-zinc at level of 100 µg L-1 In group received nano-zinc at level of 100 µg L-1, an increase in hsp70 gene expression was found. In conclusion, nano-zinc oxide at level of 100 µg L-1 could increase queen and worker performance and heat resistance of bees in the hot climate condition.

Blood lipid components and SREBP-1 gene expression in broiler chickens fed different dietary lipid sources

Background: Liver plays an important role in energy homeostasis. Intense liver diseases are accompanied with lower concentrations of n-3 and n-6 poly unsaturated fatty acids (PUFA). It has been found that n-3 PUFA play importantly protective roles in the liver. There was limited information about the effects of lipid sources on serum lipid components and liver sterol regulatory element binding-1 (SREBP-1) gene expression. Therefore, the aim of this study was to evaluate and compare the effects of fish oil, corn oil, olive oil and tallow, respectively, as dietary sources of n-3, n-6, n-9 and saturated fatty acid on serum lipid compound and liver SREBP-1 gene expression in broiler chicks.Materials, Methods & Results: In a completely randomized design, 240 one-day-old broiler chicks were allocated to five dietary treatments, four replicates and 12 chicks per each. Dietary treatments included of: control (diet without lipid supplementation) and diet supplemented with fish oil as a n-3 fatty acid source, corn oil as a n-6 fatty acid source, olive oil as a n-9 fatty acid source and tallow as a saturated fatty acid which were added to diets at 1.5, 3 and 4% in the starter, grower and finisher, respectively. At days 28 and 42 of age, liver tissue was dissected out and samples were placed in liquid nitrogen, also blood samples were collected. The SREBP-1 mRNA expression in liver [...](AU)

Genetic analysis of fertility traits of Holstein dairy cattle in warm and temperate climate / Análise genética de traços de fertilidade em vacas leiteiras Holstein em climas quentes e temperados

The edited data set for the estimation of heritability, genetic and phenotypic correlations of fertility traits contained up to 23,402 records from 10,894 cows calved between 2001 and 2015. Heritability estimates for success in first service (FS), gestation length (GL), number of inseminations (NI), insemination outcome (IO), calving interval (CI), calving birth weight (CBW) and days open (DO) were low and ranged between 0.016 (DO) and 0.123 (GL). Repeatability of fertility traits was estimated to vary from 0.021 (FS) to 0.411 (IO). The genetic correlations between DO × CI, DO × NI and CI × NI were positive and nearly perfect (0.98, 0.88 and 0.88, respectively), while those between DO × IO and CI × IO were negative (-0.98 and -1, respectively). Further, the phenotypic correlations between DO × CI, DO × NI, CI × NI, CBW × IO and SF × IO were 0.99, 0.83, 0.83, 0.99 and 1, respectively, while those between DO × IO, CI × IO, GL × IO and NI × IO were -0.99, -0.99, -0.99 and -1, respectively. Overall genetic parameters imply a good practical management in heat stress conditions will be essential for improving fertility efficiency.(AU)

Os dados editados para definir a estimativa de herdabilidade, correlações genéticas e fenotípicas de características de fertilidade continham até 23,402 registros a partir de 10,894 vacas paridas entre 2001 e 2015. As estimativas de herdabilidade para o sucesso no primeiro serviço (SF), duração da gestação (GL), número de inseminações (NI), resultado de inseminação (IO), intervalo entre partos (CI), peso ao nascer (CBW) e dias abertos (DO) foram baixas e variaram entre 0,016 (DO) e 0,123 (GL). A repetitividade das características de fertilidade foi estimada e variou entre 0,021 (SF) e 0,411 (IO). A correlação genética entre DO × CI, DO × NI e CI × NI foi positiva e quase perfeita (0,98, 0,88 e 0,88, respectivamente), enquanto que aquela entre DO × IO e CI × IO foi negativa (-0,98 e -1, respectivamente). A correlação fenotípica entre DO × CI, DO × NI, CI × NI, CBW × IO e SF × IO foi 0,99, 0,83, 0,83, 0,99 e 1, respectivamente, enquanto aquela entre DO × IO, CI × IO, GL × IO e NI × IO foi -0,99, -0,99, -0,99 e -1, respectivamente. Os parâmetros genéticos constatados implicam que será essencial uma gestão bem prática na condição de estresse por calor para melhoria da eficiência da fertilidade.(AU)

Genetic and phenotypic trends of fertility traits for Holstein dairy population in warm and temperate climate

The main objective of this study was to investigate genetic and phenotypic trends for fertility traits in Holstein dairy population under warm and temperate climate. Fertility traits were: success in first service, gestation length, number of inseminations, insemination outcome, calving interval, calving birth weight and days open. The edited data set included up to 23,402 records from 9,486 cows. The mean and standard deviation for fertility traits were 0.32 ± 0.003, 278.2 ± 5.58, 2.73 ± 1.94,0.31 ± 0.001, 415.99 ± 79.62, 40.4 ± 6.08 and 140.36 ± 76.16 for success in first service, gestation length, number of inseminations, insemination outcome, calving interval, calving birth weight and days open, respectively. In general, there were decreasing genetic trends for all traits over the years. On the other hand, there were decreasing phenotypic trend for days open, calving interval, gestation length, number of inseminations and calving birth weight, but estimates of phenotypic trends were positive for success in first service and insemination outcome over the years. It was concluded decreased trend for days open, calving interval, gestation length, number of inseminations and calving birth weight and increased trend for success in first service and insemination outcome traits over time indicated that Holstein dairy producers in warm and temperate climate were successful in managing and improving in nutrition during 1999 to 2013> (AU)

Blood lipid components and SREBP-1 gene expression in broiler chickens fed different dietary lipid sources

Background: Liver plays an important role in energy homeostasis. Intense liver diseases are accompanied with lower concentrations of n-3 and n-6 poly unsaturated fatty acids (PUFA). It has been found that n-3 PUFA play importantly protective roles in the liver. There was limited information about the effects of lipid sources on serum lipid components and liver sterol regulatory element binding-1 (SREBP-1) gene expression. Therefore, the aim of this study was to evaluate and compare the effects of fish oil, corn oil, olive oil and tallow, respectively, as dietary sources of n-3, n-6, n-9 and saturated fatty acid on serum lipid compound and liver SREBP-1 gene expression in broiler chicks.Materials, Methods & Results: In a completely randomized design, 240 one-day-old broiler chicks were allocated to five dietary treatments, four replicates and 12 chicks per each. Dietary treatments included of: control (diet without lipid supplementation) and diet supplemented with fish oil as a n-3 fatty acid source, corn oil as a n-6 fatty acid source, olive oil as a n-9 fatty acid source and tallow as a saturated fatty acid which were added to diets at 1.5, 3 and 4% in the starter, grower and finisher, respectively. At days 28 and 42 of age, liver tissue was dissected out and samples were placed in liquid nitrogen, also blood samples were collected. The SREBP-1 mRNA expression in liver [...]

Genetic analysis of fertility traits of Holstein dairy cattle in warm and temperate climate / Análise genética de traços de fertilidade em vacas leiteiras Holstein em climas quentes e temperados

The edited data set for the estimation of heritability, genetic and phenotypic correlations of fertility traits contained up to 23,402 records from 10,894 cows calved between 2001 and 2015. Heritability estimates for success in first service (FS), gestation length (GL), number of inseminations (NI), insemination outcome (IO), calving interval (CI), calving birth weight (CBW) and days open (DO) were low and ranged between 0.016 (DO) and 0.123 (GL). Repeatability of fertility traits was estimated to vary from 0.021 (FS) to 0.411 (IO). The genetic correlations between DO × CI, DO × NI and CI × NI were positive and nearly perfect (0.98, 0.88 and 0.88, respectively), while those between DO × IO and CI × IO were negative (-0.98 and -1, respectively). Further, the phenotypic correlations between DO × CI, DO × NI, CI × NI, CBW × IO and SF × IO were 0.99, 0.83, 0.83, 0.99 and 1, respectively, while those between DO × IO, CI × IO, GL × IO and NI × IO were -0.99, -0.99, -0.99 and -1, respectively. Overall genetic parameters imply a good practical management in heat stress conditions will be essential for improving fertility efficiency.

Os dados editados para definir a estimativa de herdabilidade, correlações genéticas e fenotípicas de características de fertilidade continham até 23,402 registros a partir de 10,894 vacas paridas entre 2001 e 2015. As estimativas de herdabilidade para o sucesso no primeiro serviço (SF), duração da gestação (GL), número de inseminações (NI), resultado de inseminação (IO), intervalo entre partos (CI), peso ao nascer (CBW) e dias abertos (DO) foram baixas e variaram entre 0,016 (DO) e 0,123 (GL). A repetitividade das características de fertilidade foi estimada e variou entre 0,021 (SF) e 0,411 (IO). A correlação genética entre DO × CI, DO × NI e CI × NI foi positiva e quase perfeita (0,98, 0,88 e 0,88, respectivamente), enquanto que aquela entre DO × IO e CI × IO foi negativa (-0,98 e -1, respectivamente). A correlação fenotípica entre DO × CI, DO × NI, CI × NI, CBW × IO e SF × IO foi 0,99, 0,83, 0,83, 0,99 e 1, respectivamente, enquanto aquela entre DO × IO, CI × IO, GL × IO e NI × IO foi -0,99, -0,99, -0,99 e -1, respectivamente. Os parâmetros genéticos constatados implicam que será essencial uma gestão bem prática na condição de estresse por calor para melhoria da eficiência da fertilidade.

Genetic and phenotypic trends of fertility traits for Holstein dairy population in warm and temperate climate

The main objective of this study was to investigate genetic and phenotypic trends for fertility traits in Holstein dairy population under warm and temperate climate. Fertility traits were: success in first service, gestation length, number of inseminations, insemination outcome, calving interval, calving birth weight and days open. The edited data set included up to 23,402 records from 9,486 cows. The mean and standard deviation for fertility traits were 0.32 ± 0.003, 278.2 ± 5.58, 2.73 ± 1.94,0.31 ± 0.001, 415.99 ± 79.62, 40.4 ± 6.08 and 140.36 ± 76.16 for success in first service, gestation length, number of inseminations, insemination outcome, calving interval, calving birth weight and days open, respectively. In general, there were decreasing genetic trends for all traits over the years. On the other hand, there were decreasing phenotypic trend for days open, calving interval, gestation length, number of inseminations and calving birth weight, but estimates of phenotypic trends were positive for success in first service and insemination outcome over the years. It was concluded decreased trend for days open, calving interval, gestation length, number of inseminations and calving birth weight and increased trend for success in first service and insemination outcome traits over time indicated that Holstein dairy producers in warm and temperate climate were successful in managing and improving in nutrition during 1999 to 2013>

The impacts of prebiotic and feed restriction on serum biochemical parameters of broilers under microbial stress caused by Salmonella enterica

Background: Feed restriction is limiting feed in one period of breeding and has advantages such as decreasing ascite and sudden death syndrome. During feed restriction, colonization of pathogens . A great number of researches have shown the positive effects of feed restriction or prebiotic on performance in separate, but few have been done on their effects on blood parameters, especially in pathogen challenging condition. The aim of this study was to investigate the impacts of prebiotic and feed restriction, alone and together, on serum biochemical parameters of broilers under microbial stress caused by Salmonella enterica challenge.Materials, Methods & Results: A total of 320 male broilers (Ross 308) at a2×2×2 factorial arrangement in randomized complete block design were divided into eight treatment groups with four replicates per each. The impact of main effects of prebiotic, feed restriction and Salmonella challenge and their interactions on serum parameters were measured at days 21 and 42 of age. The addition of prebiotic to diet decreased serum cholesterol (P 0.05).[...]

The impacts of prebiotic and feed restriction on serum biochemical parameters of broilers under microbial stress caused by Salmonella enterica

Background: Feed restriction is limiting feed in one period of breeding and has advantages such as decreasing ascite and sudden death syndrome. During feed restriction, colonization of pathogens . A great number of researches have shown the positive effects of feed restriction or prebiotic on performance in separate, but few have been done on their effects on blood parameters, especially in pathogen challenging condition. The aim of this study was to investigate the impacts of prebiotic and feed restriction, alone and together, on serum biochemical parameters of broilers under microbial stress caused by Salmonella enterica challenge.Materials, Methods & Results: A total of 320 male broilers (Ross 308) at a2×2×2 factorial arrangement in randomized complete block design were divided into eight treatment groups with four replicates per each. The impact of main effects of prebiotic, feed restriction and Salmonella challenge and their interactions on serum parameters were measured at days 21 and 42 of age. The addition of prebiotic to diet decreased serum cholesterol (P < 0.01) and low density lipoprotein (P < 0.01), but increased high density lipoprotein (P < 0.05), total protein (P < 0.05) and albumin (P < 0.05) at day 42 of age. The feed restriction increased triglyceride (P < 0.001) at day 21 of age. The Salmonella challenge decreased total protein (P < 0.01) and albumin (P < 0.01) at day 21 of age. Interaction between prebiotic and feed restriction on serum high density lipoprotein (P < 0.05) at day 42 of age, and the interaction between feed restriction and Salmonella challenge on serum high density lipoprotein (P < 0.05) and interaction among prebiotic, feed restriction and Salmonella challenge on serum glucose (P < 0.05) at day 21 of age were significant. Serum uric acid was not affected by the main effects and their interactions (P > 0.05).[...](AU)