Expression analysis of m6A-related genes in various tissues of Meishan pigs at different developmental stages

To characterize the N6-methyladenosine (m6A)-related gene expression profiles in various tissues of Meishan pigs at different stages, m6A modification-related genes (METTL3, METTL14, METTL16, WTAP, RBM15, and FTO) were detected from newborn to physical maturity of Meishan pigs at eight important developmental stages (1, 7, 14, 21, 28, 35, 134, and 158 days old). The expression of m6A-related genes was tissue-specific. Furthermore, the level of METTL3 messenger RNA (mRNA) was higher on day 35 than in other stages in most tissues, and the expression of METTL14 increased after day 35, and FTO exhibited a peak on day 14 in muscle, intestine, lymph nodes, thymus, and kidney. This study provided a reference for an in-depth study of the expression patterns of m6A modification-related genes in Meishan pigs.

Study on the Major Genes Related with Fat Deposition in Liver and Abdominal Fat of Different Breeds of Chicken

Fat deposition is higher in fast growing chickens than in slow growing chickens. The liver is the major organ for lipogenesis and fat deposition in chickens, although genetic background, age, and gender also influence fat deposition. In the present study, we aimed to explore the molecular mechanisms underlying fat deposition in liver and abdominal fat. We determined the expression abundances of the key genes regulating fat metabolism in fast-growing (FG) broilers (Cobb) and slow-growing (SG) broilers (HS1) and found that ACC, FAS, PGC-1α, PPARγ, SREBP-1c and PLIN1genes were expressed in the abdominal fat and liver tissues of FG and SG. ANOVA analysis showed that the breed, age, and tissue factors influenced the expressions of ACC, FAS, PGC-1α, PPARγ, SREBP-1c, and PLIN1 genes in the liver and abdominal fat of FG and SG. Also, the expressions of PPARγ and PLIN1 in the liver of SG were higher than that of FG. The results suggest that the differences in adipocyte development and adipose deposition between breeds are due to genetic factors.(AU)

Reference gene stability in the Longissimus thoracis et lumborum muscle of Colombian Creole sheep / Estabilidade de genes de referência no músculo Longissimus thoracis et lumborum de ovelhas crioulas colombianas

The Colombian Creole sheep breed has a high economic and social importance for Colombia. Both males and females of this breed are multipurpose animals, and evaluating the production and meat quality of both sexes is important for small farmers in Colombia. This requires the use of tools that help to evaluate critical production points, such as Real-time quantitative polymerase chain reaction (RT-qPCR), which is a widely used molecular tool for the relative quantification of candidate genes in various tissues. For its correct use, the use of housekeeping genes with stable expression, so-called "reference genes", is required. However, recent studies have shown that the expression of these reference genes can vary among tissues and can be modulated by breed, sex, or external stimuli. Likewise, there is little information regarding the expression of these genes in the Longissimus thoracis et lumborum muscle of male and female Colombian Creole sheep. In this study, the stability in the expression of seven reference genes (ACTB, YWHAZ, SDHA, GAPDH, TUBB2A, B2M, and PGK1) in the Longissimus thoracis et lumborum muscle of male and female Colombian Creole sheep was compared since they are used in RT-qPCR studies to determine the most stable ones for this breed. Twelve animals, six males and six females, with a body weight of 26 ± 4 kg and 12 ± 3 months of age, were used under grazing conditions. Biopsies of the Longissimus thoracis et lumborum muscle were taken, from which RNA was extracted and cDNA was synthesized. Expression was determined using RT-qPCR, and its stability was analyzed by computational algorithms using the geNorm, Normfinder, and BestKeeper software packages, which were integrated using the RefFinder software package. The results indicate that GAPDH, ACTB, and SDHA have the highest stability, whereas the most variable expression was found for B2M. These data provide the basis for more precise results in RT-qPCR studies of gene expression in the muscle of Colombian Creole sheep.(AU)

A raça de ovinos Crioula Colombiana tem grande importância econômica e social para a Colômbia. Avaliar a produção e a qualidade da carne de machos e fêmeas é importante para pequenos produtores do país e, assim, faz necessário o uso ferramentas que ajudam a avaliar os pontos críticos de produção, como a reação em cadeia da polimerase em tempo real (Real-time quantitative polymerase chain reaction [RT-qPCR]). Esta é uma ferramenta molecular amplamente usada para a quantificação relativa de genes candidatos em vários tecidos. Para o seu uso correto, é necessário o uso de genes com expressão estável denominados genes de referência. No entanto, estudos recentes têm mostrado que a expressão desses genes de referência pode variar entre os tecidos e pode ser modulada por raça, sexo ou estímulos externos. Da mesma forma, existem poucas informações sobre a expressão desses genes no músculo Longissimus thoracis et lumborum de ovinos machos e fêmeas da raça Crioula Colombiana. Neste estudo foi comparada a estabilidade na expressão de sete genes de referência (ACTB, YWHAZ, SDHA, GAPDH, TUBB2A, B2M e PGK1) no músculo Longissimus thoracis et lumborum de ovinos Crioulo Colombiano machos e fêmeas, por serem genes utilizados em estudos de RT-qPCR visando determinar os mais estáveis para esta raça. Doze animais com peso corporal de 26 ± 4 kg e 12 ± 3 meses de idade foram utilizados em condições de pastejo. Foram realizadas biópsias do músculo Longissimus thoracis et lumborum, de onde o RNA foi extraído e o cDNA foi sintetizado. A expressão foi determinada usando RT-qPCR e sua estabilidade foi analisada por algoritmos computacionais usando geNorm, Normfinder e BestKeeper pacote de software, os quais foram integrados usando RefFinder pacote de software. Os resultados indicam que GAPDH, ACTB e SDHA apresentam maior estabilidade, enquanto a expressão mais variável foi para B2M. Esses dados fornecem a base para resultados mais precisos em estudos de RT-qPCR de expressão gênica em músculos defeminino ovinos da raça Crioula Colombiana.(AU)

Detection of Fur, AmoA and pvcAB genes in Aeromonas hydrophila isolated from aquatic organisms and impact on bacterial growth under different iron concentrations / Detecção dos genes Fur, AmoA e pvcAB em Aeromonas hydrophila isoladas de organismos aquáticos e o impacto da presença desses genes sobre o crescimento bacteriano com diferentes concentrações de ferro

Infection caused by Aeromonas brings great harm to fish farming. Among the factors associated with bacterial pathogenesis, iron uptake can contribute to the survival and virulence of bacteria within hosts. The aim of this study was to check the presence of genes related to iron uptake in Aeromonas hydrophila deriving from aquatic organisms in the São Francisco Valley and associate the presence of these genes with the ability to grow in media containing different concentrations of iron. The DNAs of 41 isolates were extracted and used in PCRs to verify the presence of the Fur, AmoA and pvcAB genes related to iron uptake. The growth of the isolates belonging to different genetic profiles was verified in culture media containing different iron concentrations. Two isolates were positive for the presence of the Fur gene, seven for the AmoA gene and two for the pvcAB gene. The growth test showed that the low availability of iron did not interfere in the growth of the isolates, nor in the isolate that did not contain any of the genes evaluated in this study, suggesting that the iron uptake's mechanisms of the tested isolates may be related to other genes and proteins.

Infecções causadas por Aeromonas trazem grandes prejuízos à piscicultura. Entre os fatores associados à patogênese bacteriana, a captação de ferro pode contribuir para a sobrevivência e a virulência das bactérias dentro dos hospedeiros. O objetivo deste estudo foi verificar a presença de genes relacionados à captação de ferro em Aeromonas hydrophila provenientes de organismos aquáticos do Vale do São Francisco e associar a presença desses genes com a capacidade de as bactérias crescerem em meios contendo diferentes concentrações de ferro. Os DNAs de 41 isolados foram extraídos e utilizados em PCRs para verificar a presença dos genes Fur, AmoA e pvcAB relacionados à captação de ferro. O crescimento dos isolados pertencentes a diferentes perfis genéticos foi verificado em meios de cultura contendo diferentes concentrações de ferro. Dois isolados foram positivos para a presença do gene Fur, sete para a do gene AmoA e dois para a do gene pvcAB. O teste de crescimento mostrou que a baixa disponibilidade de ferro não interferiu no crescimento dos isolados nem no isolado que não continha nenhum dos genes avaliados neste estudo, sugerindo que os mecanismos de captação de ferro dos isolados testados podem estar relacionados a outros genes e proteínas.

Expression of icaA and icaD genes in biofilm formation in Staphylococcus aureus isolates from bovine subclinical mastitis / Expressão dos genes icaA e icaD na formação de biofilme em isolados de Staphylococcus aureus de mastite subclínica bovina

Staphylococcus spp. plays a significant role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most important species due to the high prevalence and the difficulty of in vivo treatment that is related to the expression of virulence factors and biofilm formation. This study aimed to detect the phenotypic expression of the biofilm formation in 20 S. aureus isolated from bovine mastitis and to evaluate the expression and regulation of genes involved in its production. MALDI-TOF and phenogenotypic identification assays were performed to characterize the isolates. The phenotypic biofilm production and the presence of icaA and icaD and bap genes were evaluated. The Agr system was typified (agr I, agr II, agr III and agr IV) and its regulator (agr RNAIII) was detected. Furtherly, Real-time PCR (qPCR) was performed at chosen times to quantify the expression of icaA, icaD and hld genes in three selected isolates. All 20 strains were biofilm producers and most presented icaA and icaD genes. Only one isolate presented the bap gene. The agr gene type II showed a prevalence of 70%. Transcriptional analysis revealed increased expression of ica genes at eight hours of growth. These results confirm that polysaccharides production mediated by the icaADBC operon genes is an essential mechanism to the biofilm formation and contributes to the early stages of bacterial growth.(AU)

Staphylococcus spp. desempenham um papel significativo na etiologia da mastite bovina. Staphylococcus aureus é considerada a espécie mais importante devido a alta prevalência e a dificuldade de tratamento in vivo que está relacionado à expressão dos fatores de virulência e formação de biofilme. Este estudo teve como objetivo detectar a expressão fenotípica da formação de biofilme em 20 cepas de S. aureus isoladas de mastite bovina e avaliar a expressão e regulação de genes envolvidos em sua produção. MALDI-TOF e ensaios de identificação fenogenotípica foram realizados para caracterizar os isolados. A produção fenotípica de biofilme e a presença dos genes icaA, icaD e bap foram avaliadas. O sistema Agr foi tipificado (agr I, agr II, agr III e agr IV) e seu regulador (agr RNAIII) foi detectado. Além disso, a PCR em tempo real (qPCR) foi realizada nos tempos determinados para quantificar a expressão dos genes icaA, icaD e hld em três isolados selecionados. Todas as 20 linhagens foram produtoras de biofilme e a maioria apresentava os genes icaA e icaD. Apenas um isolado apresentou o gene bap. O gene agr do tipo II mostrou uma prevalência de 70%. A análise transcricional revelou aumento da expressão de genes ica às oito horas de crescimento. Estes resultados confirmam que a produção de polissacarídeos mediada pelos genes do operon icaADBC é um mecanismo essencial para a formação do biofilme e contribui para os estágios iniciais do crescimento bacteriano.(AU)

Expression of icaA and icaD genes in biofilm formation in Staphylococcus aureus isolates from bovine subclinical mastitis / Expressão dos genes icaA e icaD na formação de biofilme em isolados de Staphylococcus aureus de mastite subclínica bovina

Staphylococcus spp. plays a significant role in the etiology of bovine mastitis. Staphylococcus aureus is considered the most important species due to the high prevalence and the difficulty of in vivo treatment that is related to the expression of virulence factors and biofilm formation. This study aimed to detect the phenotypic expression of the biofilm formation in 20 S. aureus isolated from bovine mastitis and to evaluate the expression and regulation of genes involved in its production. MALDI-TOF and phenogenotypic identification assays were performed to characterize the isolates. The phenotypic biofilm production and the presence of icaA and icaD and bap genes were evaluated. The Agr system was typified (agr I, agr II, agr III and agr IV) and its regulator (agr RNAIII) was detected. Furtherly, Real-time PCR (qPCR) was performed at chosen times to quantify the expression of icaA, icaD and hld genes in three selected isolates. All 20 strains were biofilm producers and most presented icaA and icaD genes. Only one isolate presented the bap gene. The agr gene type II showed a prevalence of 70%. Transcriptional analysis revealed increased expression of ica genes at eight hours of growth. These results confirm that polysaccharides production mediated by the icaADBC operon genes is an essential mechanism to the biofilm formation and contributes to the early stages of bacterial growth.(AU)

Staphylococcus spp. desempenham um papel significativo na etiologia da mastite bovina. Staphylococcus aureus é considerada a espécie mais importante devido a alta prevalência e a dificuldade de tratamento in vivo que está relacionado à expressão dos fatores de virulência e formação de biofilme. Este estudo teve como objetivo detectar a expressão fenotípica da formação de biofilme em 20 cepas de S. aureus isoladas de mastite bovina e avaliar a expressão e regulação de genes envolvidos em sua produção. MALDI-TOF e ensaios de identificação fenogenotípica foram realizados para caracterizar os isolados. A produção fenotípica de biofilme e a presença dos genes icaA, icaD e bap foram avaliadas. O sistema Agr foi tipificado (agr I, agr II, agr III e agr IV) e seu regulador (agr RNAIII) foi detectado. Além disso, a PCR em tempo real (qPCR) foi realizada nos tempos determinados para quantificar a expressão dos genes icaA, icaD e hld em três isolados selecionados. Todas as 20 linhagens foram produtoras de biofilme e a maioria apresentava os genes icaA e icaD. Apenas um isolado apresentou o gene bap. O gene agr do tipo II mostrou uma prevalência de 70%. A análise transcricional revelou aumento da expressão de genes ica às oito horas de crescimento. Estes resultados confirmam que a produção de polissacarídeos mediada pelos genes do operon icaADBC é um mecanismo essencial para a formação do biofilme e contribui para os estágios iniciais do crescimento bacteriano.(AU)

Detection of resistance genes in pyometra isolated bacteria in bitches / Detecção de genes de resistência em bactérias isoladas de piometra em cadelas

Pyometra has several immunological and molecular changes that are responsible for uterine inflammation and the disease may or may not have infections. This study aimed to isolate and identify bacteria in the uterine content of bitches with pyometra, to analyze the susceptibility profile to antibiotics, detect ß-lactamase enzyme production by phenotypic tests, and resistance genes to ß-lactams. Eighteen samples of uterine content were collected by aspiration puncture. The samples were inoculated in bacteriological media and identified by biochemical tests. Subsequently, antibiogram tests, screening for detection of ß-lactamases, and Real-Time PCR for detection of resistance genes was performed. Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Citrobacter spp., Staphylococcus spp., and Streptococcus spp. were identified in the analyzed samples of uterine content. In the antibiogram test, 90.5% of the isolates showed resistance to at least one antibiotic, and of these, 36.8% were considered MDR, with three Staphylococcus spp., three E. coli, and one Klebsiellaspp. Concerning bacterial resistance to the groups of antibiotics tested, 38.1% of the isolates were resistant to at least one type of ß-lactam, 33.3% to tetracycline, 19.0% to aminoglycosides, and 14.3% to fluoroquinolones, macrolides, and trimethoprim-sulfamethoxazole. In the phenotypic test to detect ß-lactamase production, E. coli samples were negative and Klebsiella spp. was positive for the production of AmpC, which presented the blaCMY, blaSPM, and blaSIM genes. Bacteria that are resistant to antibiotics represent a great challenge and laboratory support is therefore essential, without which therapeutic success decreases and death may be inevitable.(AU)

A piometra apresenta diversas alterações imunológicas e moleculares que são responsáveis pela inflamação uterina, e a doença pode ser infecciosa ou não. O objetivo deste estudo foi isolar e identificar bactérias no conteúdo uterino de cadelas com piometra, analisar o perfil de suscetibilidade aos antibióticos, detectar a produção de enzimas ß-lactamase por testes fenotípicos e genes de resistência aos ß-lactâmicos. Dezoito amostras de conteúdo uterino foram coletadas por punção aspirativa. As amostras foram inoculadas em meio bacteriológico e identificadas por testes bioquímicos. Posteriormente, foram realizados testes de antibiograma, triagem para detecção de ß-lactamases e PCR em tempo real para detecção de genes de resistência. Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Citrobacter spp., Staphylococcus spp. e Streptococcus spp. foram identificados nas amostras de conteúdo uterino analisadas. No teste de antibiograma, 90,5% dos isolados apresentaram resistência a pelo menos um antibiótico, e destes, 36,8% foram considerados MR, sendo três Staphylococcus spp., três E. coli e uma Klebsiella spp. Sobre a resistência bacteriana aos grupos de antibióticos testados, 38,1% dos isolados foram resistentes a pelo menos um tipo de ß-lactâmico, 33,3% à tetraciclina, 19,0% aos aminoglicosídeos e 14,3% às fluorquinolonas, macrolídeos e trimetoprim-sulfametoxazol. No teste fenotípico para detecção da produção de ß-lactamase, as amostras de E. coli foram negativas, e Klebsiella spp. foi positiva para a produção de AmpC, que apresentou os genes blaCMY, blaSPM e blaSIM. As bactérias resistentes aos antibióticos representam um grande desafio e, portanto, o suporte laboratorial é essencial, sem o qual o sucesso terapêutico diminui e a morte pode ser inevitável.(AU)

Detection of resistance genes in pyometra isolated bacteria in bitches / Detecção de genes de resistência em bactérias isoladas de piometra em cadelas

Pyometra has several immunological and molecular changes that are responsible for uterine inflammation and the disease may or may not have infections. This study aimed to isolate and identify bacteria in the uterine content of bitches with pyometra, to analyze the susceptibility profile to antibiotics, detect ß-lactamase enzyme production by phenotypic tests, and resistance genes to ß-lactams. Eighteen samples of uterine content were collected by aspiration puncture. The samples were inoculated in bacteriological media and identified by biochemical tests. Subsequently, antibiogram tests, screening for detection of ß-lactamases, and Real-Time PCR for detection of resistance genes was performed. Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Citrobacter spp., Staphylococcus spp., and Streptococcus spp. were identified in the analyzed samples of uterine content. In the antibiogram test, 90.5% of the isolates showed resistance to at least one antibiotic, and of these, 36.8% were considered MDR, with three Staphylococcus spp., three E. coli, and one Klebsiellaspp. Concerning bacterial resistance to the groups of antibiotics tested, 38.1% of the isolates were resistant to at least one type of ß-lactam, 33.3% to tetracycline, 19.0% to aminoglycosides, and 14.3% to fluoroquinolones, macrolides, and trimethoprim-sulfamethoxazole. In the phenotypic test to detect ß-lactamase production, E. coli samples were negative and Klebsiella spp. was positive for the production of AmpC, which presented the blaCMY, blaSPM, and blaSIM genes. Bacteria that are resistant to antibiotics represent a great challenge and laboratory support is therefore essential, without which therapeutic success decreases and death may be inevitable.(AU)

A piometra apresenta diversas alterações imunológicas e moleculares que são responsáveis pela inflamação uterina, e a doença pode ser infecciosa ou não. O objetivo deste estudo foi isolar e identificar bactérias no conteúdo uterino de cadelas com piometra, analisar o perfil de suscetibilidade aos antibióticos, detectar a produção de enzimas ß-lactamase por testes fenotípicos e genes de resistência aos ß-lactâmicos. Dezoito amostras de conteúdo uterino foram coletadas por punção aspirativa. As amostras foram inoculadas em meio bacteriológico e identificadas por testes bioquímicos. Posteriormente, foram realizados testes de antibiograma, triagem para detecção de ß-lactamases e PCR em tempo real para detecção de genes de resistência. Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Citrobacter spp., Staphylococcus spp. e Streptococcus spp. foram identificados nas amostras de conteúdo uterino analisadas. No teste de antibiograma, 90,5% dos isolados apresentaram resistência a pelo menos um antibiótico, e destes, 36,8% foram considerados MR, sendo três Staphylococcus spp., três E. coli e uma Klebsiella spp. Sobre a resistência bacteriana aos grupos de antibióticos testados, 38,1% dos isolados foram resistentes a pelo menos um tipo de ß-lactâmico, 33,3% à tetraciclina, 19,0% aos aminoglicosídeos e 14,3% às fluorquinolonas, macrolídeos e trimetoprim-sulfametoxazol. No teste fenotípico para detecção da produção de ß-lactamase, as amostras de E. coli foram negativas, e Klebsiella spp. foi positiva para a produção de AmpC, que apresentou os genes blaCMY, blaSPM e blaSIM. As bactérias resistentes aos antibióticos representam um grande desafio e, portanto, o suporte laboratorial é essencial, sem o qual o sucesso terapêutico diminui e a morte pode ser inevitável.(AU)

Herbicide management in glyphosate and sulfonylurea-tolerant soybeans / Manejo de herbicidas em soja tolerante a glifosato e sulfonilureias

There is little information on the efficacy and selectivity of sulfonylureas, isolated and in association with glyphosate, in glyphosate and sulfonylurea-tolerant soybeans. Thus, the present study aims to evaluate the efficacy of weed control and selectivity of sulfonylureas, isolated and in association with glyphosate, at post-emergence (V4) of RR2/STS soybean. The experiments were conducted in the in areas located in Piracicaba City, São Paulo State (SP), Brazil (experiment I) and Palotina City, Paraná State (PR), Brazil (experiment II). Treatments were composed of application of the herbicides sulfometuron, chlorimuron, halosulfuron, ethoxysulfuron and glyphosate, isolated and in association, in the BMX Garra RR2/STS cultivar. Experiment I was conducted focusing on the evaluation of the efficacy of weed control; whereas experiment II focused mainly on the evaluation of herbicide selectivity. The experimental design was the randomized complete block, with four replications. Crop injury, weed control, and variables related to agronomic performance were evaluated. Data were submitted to analysis of variance, and the means of the treatments were compared with the Tukey test. Sulfonylureas in association with glyphosate were effective in weed control and selective for the BMX Garra RR2/STS soybean cultivar. The sulfometuron + chlorimuron + glyphosate association presented phytotoxic potential for the BMX Garra RR2/STS cultivar.(AU)

Há poucas informações sobre a eficácia e seletividade de sulfonilureias, isoladas e associadas ao glifosato, na soja tolerante ao glifosato e às sulfonilureias. Assim, o presente estudo teve como objetivo avaliar a eficácia no controle de plantas daninhas e seletividade de sulfonilureias, isoladas e em associação com o glifosato, em pós-emergência (V4) de soja RR2/STS. Os experimentos foram conduzidos em áreas localizadas nos municípios de Piracicaba, São Paulo (SP), Brasil (experimento I) e Palotina, Paraná (PR), Brasil (experimento II). Os tratamentos foram compostos pela aplicação dos herbicidas sulfometurom, clorimurom, halossulfurom, etoxissulfurom e glifosato, isolados e em associação, no cultivar BMX Garra RR2/STS. O experimento I foi realizado com o foco principal na avaliação da eficácia no controle de plantas daninhas; ao passo que o experimento II se concentrou principalmente na avaliação da seletividade dos herbicidas. O delineamento experimental foi o de blocos casualizados, com quatro repetições. Foram avaliados sintomas de injúria, controle de plantas daninhas e variáveis relacionadas ao desempenho agronômico. Os dados foram submetidos à análise de variância, e as médias dos tratamentos foram comparadas pelo teste de Tukey. Sulfonilureias associadas ao glifosato foram eficazes no controle de plantas daninhas e seletivas para o cultivar de soja BMX Garra RR2/STS. A associação sulfometurom + clorimurom + glifosato apresentou potencial fitotóxico para o cultivar BMX Garra RR2/STS.(AU)

Herbicide management in glyphosate and sulfonylurea-tolerant soybeans / Manejo de herbicidas em soja tolerante a glifosato e sulfonilureias

There is little information on the efficacy and selectivity of sulfonylureas, isolated and in association with glyphosate, in glyphosate and sulfonylurea-tolerant soybeans. Thus, the present study aims to evaluate the efficacy of weed control and selectivity of sulfonylureas, isolated and in association with glyphosate, at post-emergence (V4) of RR2/STS soybean. The experiments were conducted in the in areas located in Piracicaba City, São Paulo State (SP), Brazil (experiment I) and Palotina City, Paraná State (PR), Brazil (experiment II). Treatments were composed of application of the herbicides sulfometuron, chlorimuron, halosulfuron, ethoxysulfuron and glyphosate, isolated and in association, in the BMX Garra RR2/STS cultivar. Experiment I was conducted focusing on the evaluation of the efficacy of weed control; whereas experiment II focused mainly on the evaluation of herbicide selectivity. The experimental design was the randomized complete block, with four replications. Crop injury, weed control, and variables related to agronomic performance were evaluated. Data were submitted to analysis of variance, and the means of the treatments were compared with the Tukey test. Sulfonylureas in association with glyphosate were effective in weed control and selective for the BMX Garra RR2/STS soybean cultivar. The sulfometuron + chlorimuron + glyphosate association presented phytotoxic potential for the BMX Garra RR2/STS cultivar.(AU)

Há poucas informações sobre a eficácia e seletividade de sulfonilureias, isoladas e associadas ao glifosato, na soja tolerante ao glifosato e às sulfonilureias. Assim, o presente estudo teve como objetivo avaliar a eficácia no controle de plantas daninhas e seletividade de sulfonilureias, isoladas e em associação com o glifosato, em pós-emergência (V4) de soja RR2/STS. Os experimentos foram conduzidos em áreas localizadas nos municípios de Piracicaba, São Paulo (SP), Brasil (experimento I) e Palotina, Paraná (PR), Brasil (experimento II). Os tratamentos foram compostos pela aplicação dos herbicidas sulfometurom, clorimurom, halossulfurom, etoxissulfurom e glifosato, isolados e em associação, no cultivar BMX Garra RR2/STS. O experimento I foi realizado com o foco principal na avaliação da eficácia no controle de plantas daninhas; ao passo que o experimento II se concentrou principalmente na avaliação da seletividade dos herbicidas. O delineamento experimental foi o de blocos casualizados, com quatro repetições. Foram avaliados sintomas de injúria, controle de plantas daninhas e variáveis relacionadas ao desempenho agronômico. Os dados foram submetidos à análise de variância, e as médias dos tratamentos foram comparadas pelo teste de Tukey. Sulfonilureias associadas ao glifosato foram eficazes no controle de plantas daninhas e seletivas para o cultivar de soja BMX Garra RR2/STS. A associação sulfometurom + clorimurom + glifosato apresentou potencial fitotóxico para o cultivar BMX Garra RR2/STS.(AU)

Influence of temperament-related genes on live weight traits of Charolais cows

We investigated whether a panel of molecular markers previously associated with temperament has an effect on live growth traits. Phenotypic data from 412 Charolais cows categorized according to their age in adult and young cows were used to determine Pearson's correlations between birth (BW), weaning (WW), and yearling live (YW) weight and temperament traits (measured as exit velocity [EV] and temperament score [TS]). For association analysis, selective genotyping of a group of 80 cows identified as the most docile and temperamental were genotyped with a 151-SNP (single-nucleotide polymorphisms) panel of molecular markers previously associated with temperament. Significant Pearson's correlations between birth weight and weaning weight and the two temperament measurements (EV and TS) were observed only in the young cow group. Significant effects of ten SNP on BW and WW were observed. Four markers located on candidate genes for temperament traits also had an effect on birth weight and weaning weight in Charolais cows, which indicates that both traits could be influenced by the same genes.(AU)

Identification of candidate genes associated with milk yield trait in buffaloes ( Bubalus bubalis ) by restriction-site-associated DNA sequencing

The objectives of our present study included the screening of single nucleotide polymorphisms (SNP) that show significant differences in allelic frequencies between two buffalo populations (Egyptian and Chinese buffaloes), categorization of functional genes associated with these SNP by gene ontology, and pathway analyses to further understand their potential values as candidate genes closely associated with milk yield trait in buffaloes. In this study, double digest restrictionsite associated DNA sequencing was performed on Illumina HiSeq 2500 platform for 20 and 25 female buffaloes from Egypt and China, respectively. Approximately 118 Gb of sequencing data were obtained, and a total of 110,129 and 150,535 putative SNP were detected in Egyptian and Chinese buffaloes, respectively. Focused only on those SNP that differed significantly in allelic frequencies between the two populations, we found that genes associated with these SNP were significantly over-represented in the ionotropic glutamate receptor pathway, the endothelin signaling pathway, and the gonadotropin-releasing hormone receptor pathway, which contained a total of 29 genes. Of these, nine genes (ADCY5, CACNA1A, CREB1, INHBA, INHBB, PIK3R1, PLCB1, PRKCE, and SMAD2) participating in the hormonal regulation of lactation, were considered to be promising candidate genes worthy of further investigations for favorable alleles associated with milk yield. Our results provide useful information about genetic variations in Egyptian and Chinese buffaloes. The potential influences of nine candidate genes and their associated SNP on milk yield need to be validated in more buffalo populations.(AU)

Candidate genes related to reproductive traits of Hereford and Braford bulls / Genes candidatos relacionados a traços reprodutivos em touros Hereford e Braford

This study emphasized the importance of using candidate genes in predicting semen quality in bulls that can be used in cow-calf production. The aim of this study was to evaluate some candidate genes related to reproductive traits in Braford and Hereford bulls. All bulls (n=188) were submitted to breeding soundness evaluations at 24, 28, 32, and 36 months of age. The microsatellite markers ILSTS002 and BMS3004 associated with the luteinizing hormone-β (LHβ) gene, IDVGA-51 to leptin (LEP) gene,HEL5 and AFZ1 within the IGF-IR gene, and two SNP markers (LHR and FSHR) associated with the LHR and FSHR genes, respectively, were evaluated by the amplification of DNA products. The variation in the IDVGA-51 allele 177-185 showed polymorphic information content (PIC) associated with sperm motility and vigor traits in Hereford bulls. Hereford animals showed PIC of 0.36 to 0.75% along with expected heterozygosity (H) of 0.49 to 0.78%. Braford bulls that indicated the ILSTS002 allele 137-175 and AFZ1 allele 113-119 showed PIC associated with major and minor defects, respectively. The PIC ranged from 0.28 to 0.78%, with an expected H of 0.35 to 0.81%. AFZ1 allele 121-127 had the highest minor defects and ILSTS002 allele 125-135 showed the highest major defects from ejaculated semenin Braford bulls. In addition, IDVGA-51 allele 175 showed lower motility and vigor in Hereford bulls.The markers AFZ1 (IGF-IR) and ILSTS002 (LHβ) in Braford and IDVGA-51 (LEP) in Hereford bulls were effective in verifying the reproductive traits of the bulls.(AU)

Este estudo enfatizou a importância do uso de genes candidatos na predição da qualidade do sêmen em touros que podem ser utilizados na produção de bezerros. O objetivo deste estudo foi avaliar alguns genes candidatos relacionados a traços reprodutivos em touros de Braford e Hereford. Todos os touros (n = 188) foram submetidos a avaliações reprodutivas aos 24, 28, 32 e 36 meses de idade. Os marcadores microsatélites ILSTS002 e BMS3004 associados ao gene hormônio luteinizante-β (LHβ), o IDVGA-51 associado ao gene Leptina (LEP), o HEL5 e AFZ1 ao gene IGF-IR e dois marcadores SNP (LHR e FSHR) relacionados ao LHR e FSHR, respectivamente, foram avaliados pela amplificação de produtos de DNA. A variação no alelo IDVGA-51 177-185 mostrou conteúdo de informação polimórfica (PIC) associada à motilidade espermática e características de vigor nos touros de Hereford. Os animais Hereford apresentaram PIC de 0,36 a 0,75%, juntamente com heterozigosidade esperada (H) de 0,49 a 0,78%. Os touros Braford que indicaram o alelo ILSTS002 137-175 e o alelo AFZ1 113-119 mostraram PIC associados a defeitos maiores e menores, respectivamente. O PIC variou de 0,28 a 0,78% com um H esperado de 0,35 a 0,81%. O alelo AFZ1 121-127 teve os defeitos menores mais altos, enquanto, o alelo ILSTS002 125-135 mostrou altos defeitos maiores do sêmen ejaculado em Braford. Além disso, o alelo IDVGA-51 175 mostrou baixa motilidade e vigor nos touros Hereford. Os marcadores AFZ1 (IGF-IR) e ILSTS002 (LHβ) em Braford e IDVGA-51 (LEP) no Hereford foram efetivos na verificação dos traços reprodutivos dos touros.(AU)

Identification of genes associated with reproductive function in dairy cattle

The use of genomics has improved response to selection for functional traits with low heritability such as fertility traits. Much of the work on fertility traits has been performed through use of genome-wide association studies (GWAS) to identify genetic loci associated with reproductive traits. Under a GWAS approach, the assumption is that the markers on the panel are in linkage disequilibrium with causative mutations. In many cases, identification of the causative mutation is difficult because an associated genetic marker can be in intergenic regions and can be in linkage disequilibrium with variants in several nearby genes. Another approach is to identify candidate genes using knowledge of the biological pathways controlling a trait to search for single nucleotide polymorphism (SNP) in genes in those pathways. This should reveal putative causative markers responsible for genetic variation in biological function, and it is expected that the marker will be more strongly associated with a trait than one in linkage disequilibrium. An example of how a series of candidate gene studies demonstrate that identification of markers in genes involved in reproductive processes can lead to discovery of additional markers associated with genetic variation in reproductive traits is presented. In addition, the inclusion of candidate markers for fertility can improve reliability of genetic estimates for fertility traits, and the repeatability of the effects across a separate population of animals gives confidence that association elucidated by this set of markers is likely to be real. More importantly, the use of candidate genes can provide insights into the biology underpinning genetic variation in fertility, and that this understanding can lead to physiological interventions to improve reproductive function.

Identification of genes associated with reproductive function in dairy cattle

The use of genomics has improved response to selection for functional traits with low heritability such as fertility traits. Much of the work on fertility traits has been performed through use of genome-wide association studies (GWAS) to identify genetic loci associated with reproductive traits. Under a GWAS approach, the assumption is that the markers on the panel are in linkage disequilibrium with causative mutations. In many cases, identification of the causative mutation is difficult because an associated genetic marker can be in intergenic regions and can be in linkage disequilibrium with variants in several nearby genes. Another approach is to identify candidate genes using knowledge of the biological pathways controlling a trait to search for single nucleotide polymorphism (SNP) in genes in those pathways. This should reveal putative causative markers responsible for genetic variation in biological function, and it is expected that the marker will be more strongly associated with a trait than one in linkage disequilibrium. An example of how a series of candidate gene studies demonstrate that identification of markers in genes involved in reproductive processes can lead to discovery of additional markers associated with genetic variation in reproductive traits is presented. In addition, the inclusion of candidate markers for fertility can improve reliability of genetic estimates for fertility traits, and the repeatability of the effects across a separate population of animals gives confidence that association elucidated by this set of markers is likely to be real. More importantly, the use of candidate genes can provide insights into the biology underpinning genetic variation in fertility, and that this understanding can lead to physiological interventions to improve reproductive function.(AU)

New morphological data and molecular diagnostic of Henneguya friderici (Myxozoa: Myxobolidae), a parasite of Leporinus friderici (Osteichthyes: Anostomidae) from southeastern Brazil / Novos dados morfológicos e diagnóstico molecular de Henneguya friderici (Myxozoa: Myxobolidae), parasito de Leporinus friderici (Osteichthyes: Anostomidae) do sudeste do Brasil

The myxozoan Henneguya friderici is a parasite of the gills, intestine, kidney and liver of Leporinus friderici, a characiform fish belonging to the family Anostomidae. Forty-two specimens of L. friderici that had been caught in the Mogi Guaçú River, state of São Paulo, were studied. Elongated white plasmodia were found in the gill filaments of 10 host specimens (24%). The mature spores had an ellipsoidal body with polar capsules of equal size and caudal length greater than body length. This study also described 18S rDNA sequencing of H. friderici infecting the gill filaments. This produced a sequence of 1050 bp that demonstrated significant genetic differences with previously described species of Henneguya. Similarity analysis using sequences from species that clustered closest to those produced by this study showed that the species with greatest genetic similarity to H. friderici was H. leporinicola, with 94% similarity.(AU)

O myxozoa Henneguya friderici é um parasito encontrado nas brânquias, fígado, intestino e rins de Leporinus friderici, (Characiformes: Anastomidae). Foram capturados e examinados quarenta e dois espécimes de L. friderici oriundos do Rio Mogi Guaçú, estado de São Paulo. Cistos alongados e brancos foram encontrados nos filamentos branquiais de 10 (24%) hospedeiros. Os esporos maduros apresentaram o corpo alongado com as cápsulas polares em tamanhos iguais e o comprimento caudal maior do que o comprimento corporal. Com isso, o presente trabalho, descreve o sequenciamento de 1050 pb do gene 18S rDNA de H. friderici infectando os filamentos branquiais, o que demonstrou diferenças genéticas significativas em comparação com espécies previamente descritas de Henneguya. A análise de similaridade utilizando as sequencias de espécies que se agruparam mais próximas às produzidas por este estudo mostrou que a espécie com maior semelhança genética com H. friderici foi H. leporinicola, com 94% de similaridade.(AU)

Fusarium culmorum affects expression of biofilm formation key genes in Bacillus subtilis

It is known that there is correlation between biofilm formation and antagonistic activities of Bacillus subtilis strains; but, the mechanism of this correlation is not clear. So, the effect of the plant pathogen (Fusarium culmorum) on the biofilm formation in a B. subtilis strain with high antagonistic and biofilm formation activities was studied. The expression of sinR and tasA genes involved in the biofilm formation was studied in both single culture of bacterium (B) and co-culture with F. culmorum (FB) using real-time PCR. The results revealed that the expression of the sinR gene in both B and FB conditions was continuously decreased during the biofilm formation period and, after 24 h (B4 and FB4), it reached 1% and 0.3% at the planktonic phase (B1), respectively, whereas the expression of the tasA was continuously increased and was 5.27 and 30 times more than that at the planktonic phase (B1) after 24 h, respectively. So, the expression reduction rate for sinR (3 times) and the expression increasing rate for tasA (6 times) were significantly higher in FB conditions than the B ones. The relative expression of sinR in FB1 (planktonic phase), FB2 (8 h), FB3(12 h), and FB4 (24 h) times was 0.65, 0.44, 0.35, and 0.29, whereas the tasA gene expression was 2.98, 3.44, 4.37, and 5.63-fold of the one at coordinate time points in B conditions, respectively. The significant expression reduction of sinR and increase of tasA confirmed that the presence of pathogen could stimulate biofilm formation in the antagonistic bacterium. (AU)

Expression of genes SBP and leginsulin in contrasting soybean seed coats / Expressão dos genes SBP e leginsulin em tegumentos de sementes de soja contrastantes

Evaluation of differential candidate gene expression in contrasting soybean seeds is an auxiliary tool in the partial elucidation of processes involved in seeds formation, as well as it contributes to the generation of new information that can be used in future research or in the development of r genetic superior constitutions. The aim of this study was to evaluate the expression of two candidate genes, SBP and leginsulin genes, possibly involved in seed quality, in contrasting coats of four soybean genotypes. Two cultivars of yellow soybeans were used, BMX Potência RR and CD 202, and two lines of black soybean, TP and IAC. Gene expression was evaluated using qPCR in seven stages of development from seed coats for four genotypes, at 25, 30, 35, 40, 45, 50, and 55 days after anthesis. The design was completely randomized, with three replications. Data were subjected to analysis of variance and means compared by Tukey's test at 5% probability. SBP and leginsulin gene have higher expression in the early phases of development from seed coats of BMX Potência RR cultivar, followed by the IAC line. These genotypes are therefore of interest for further research involving these genes.(AU)

A avaliação da expressão diferencial de genes candidatos em sementes de soja contrastantes é uma ferramentaauxiliar na elucidação parcial dos processos envolvidos na formação de sementes, além de contribuir para a geração de novas informações que possam ser usadas em futuras pesquisas ou para o desenvolvimento de constituições genéticas superiores. O objetivo do trabalho foi avaliar a expressão de dois genes candidatos, SBP e leginsulin, possivelmente envolvidos com a qualidade das sementes, em tegumentos contrastantes de quatro genótipos de soja. Foram utilizadas duas cultivares de tegumentos amarelos, BMX Potência RR e CD 202, e duas linhagens de tegumentos pretos, TP e IAC. A expressão dos genes foi avaliada pela técnica qPCR, em sete fases de desenvolvimento dos tegumentos para os quatro genótipos, aos 25, 30, 35, 40, 45, 50 e 55 dias após a antese. O delineamento foi o inteiramente casualizado, com três repetições. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey, em nível de 5% de probabilidade. Os genes SBP e leginsulinapresentam maior expressão nas fases iniciais de desenvolvimento dos tegumentos de BMX Potência RR, seguido de IAC, sendo estes genótipos de interesse para novas pesquisas envolvendo estes genes.(AU)

Age-dependent expression of Pten and Smad4 genes in the urogenital system of Wistar rats

PURPOSE: To analyze Pten and Smad4 gene expression in the urogenital system of Wistar rats in differents ages. METHODS: Pten and Smad4 mRNA expression was assessed in the bladder, ventral prostate, testis, ovaries, and uterus by real-time PCR. Statistical analysis using the ANOVA (p<0.05). RESULTS: Pten levels showed a progressive age-dependent increase in the bladder (male and female) and prostate and were elevated in the ovaries of the middle-aged. In the uterus, no statistically significant differences were observed; in the testis, increased and decreased levels were seen in young adult and middle-aged rats, respectively. Smad4 expression was downregulated in the ovaries of the pubertal group but increased in the middle age group. In the uterus, Smad4 expression in the oldest group was higher than the others groups. In the testis, Smad4 expression steadily declined with age; in the prostate, it was higher in middle-aged rats than in younger rats. A similar trend was observed in the bladder of male and female middle-aged rats, compared with the pubertal group. CONCLUSION: The changes in phosphatase tensin homologue and Smad4 mRNA expression in Wistar rats appear to be associated with hormonal modifications in puberty and may be related to early follicular and testicular development. .(AU)

Identification and characterization of rhizospheric microbial diversity by 16S ribosomal RNA gene sequencing

In the present study, samples of rhizosphere and root nodules were collected from different areas of Pakistan to isolate plant growth promoting rhizobacteria. Identification of bacterial isolates was made by 16S rRNA gene sequence analysis and taxonomical confirmation on EzTaxon Server. The identified bacterial strains were belonged to 5 genera i.e. Ensifer, Bacillus, Pseudomona, Leclercia and Rhizobium. Phylogenetic analysis inferred from 16S rRNA gene sequences showed the evolutionary relationship of bacterial strains with the respective genera. Based on phylogenetic analysis, some candidate novel species were also identified. The bacterial strains were also characterized for morphological, physiological, biochemical tests and glucose dehydrogenase (gdh) gene that involved in the phosphate solublization using cofactor pyrroloquinolone quinone (PQQ). Seven rhizoshperic and 3 root nodulating stains are positive for gdh gene. Furthermore, this study confirms a novel association between microbes and their hosts like field grown crops, leguminous and non-leguminous plants. It was concluded that a diverse group of bacterial population exist in the rhizosphere and root nodules that might be useful in evaluating the mechanisms behind plant microbial interactions and strains QAU-63 and QAU-68 have sequence similarity of 97 and 95% which might be declared as novel after further taxonomic characterization.