Phagocytotic activity and gene expression of leukocytes isolated from Astyanax lacustris / Atividade fagocítica e expressão gênica de leucócitos isolados de Astyanax lacustris

The constant intensification of aquaculture has considerable increased the stress levels of farmed fish and, consequently, the number and intensity of diseases outbreaks. Thus, studies on fish immune response, especially regarding the interaction of fish leukocytes with potential pathogens and xenobiotics are of great importance in order to develop new prophylactic and curative strategies. We isolated leukocytes from the head kidney of Astyanax lacustris­an important Neotropical fish species for aquaculture and a potential model for Neotropical aquaculture research­using a Percoll centrifugation protocol. The isolated leukocytes were incubated with lipopolysaccharide (LPS), and the expression of genes IL-1ß, IL-8, LysC, and LysG were measured. We assessed the phagocytotic activity of leukocytes using Congo red-dyed yeast, a novel and cost-effective protocol that has been developed in this study. The isolated leukocytes responded to LPS induction, exhibiting strong IL-1ß and IL-8 upregulation, two of the most important pro-inflammatory interleukins for vertebrates immune reponse. The optimal concentration of yeast for the phagocytic assay was 106 cells mL-1, resulting in acceptable phagocytic capacity (PC) but without excess of yeasts during the counting process, ensuring a high precision and accuracy of the method. To the best of our knowledge, the present study is the first to investigate the in vitro gene expression and phagocytic activity of leukocytes isolated from A. lacustris. Our findings will serve as a reference for future studies on the immunology and toxicology of Neotropical fish.

A constante intensificação da aquicultura tem aumentado consideravelmente os níveis de estresse dos animais cultivados e, consequentemente, o número e a intensidade dos surtos de doenças. Logo, estudos sobre a resposta imune dos peixes, especialmente relacionados com a interação dos leucócitos de peixes com potenciais patógenos e xenobióticos, são de grande importância para o desenvolvimento de novas estratégias profiláticas e curativas. No presente trabalho, nós obtivemos sucesso ao isolar leucócitos oriundos do rim cranial de Astyanax lacustris ­ uma importante espécie de peixe Neotropical para a aquicultura e um modelo em potencial para pesquisas em aquicultura Neotropical ­ usando um protocolo de centrifugação com Percoll. Os leucócitos isolados foram incubados com lipossacarídeo (LPS) e, a expressão dos genes IL-1ß, IL-8, LysC, e LysG foi avaliada. Ainda, um novo protocolo para avaliação da atividade fagocítica dos leucócitos utilizando leveduras coradas com Vermelho Congo foi estabelecido. Os leucócitos isolados responderam à indução com LPS, exibindo up regulation dos genes IL-1ß e IL-8, duas das interleucinas pró-inflamatórias mais importantes para a resposta imune de vertebrados. Além do mais, a concentração ótima de leveduras para a avaliação da fagocitose foi de 106 células mL-1, resultando em uma capacidade fagocítica (PC) aceitável, mas sem excesso de leveduras durante o processo de contagem, garantindo maior precisão e eficácia do método. Até o presente momento, o presente estudo é o primeiro a investigar a expressão gênica e atividade fagocítica de leucócitos isolados de A. lacustris através da abordagem in vitro. Ainda, nossos resultados servirão de referência para futuros estudos em imunologia e toxicologia de peixes Neotropicais.

Avaliação do modelo NRC para predição do consumo de matéria seca por vacas leiteiras manejadas em pastos tropicais / NCR Model Evaluation for prediction of dry matter intake by dairy cows managed in tropical pastures

Objetivou-se avaliar o poder preditivo do modelo do National Research Council (NRC) para gado leiteiro em estimar o consumo de matéria seca (CMS) por vacas mestiças, em pastagens tropicais. Foi efetuada uma análise conjunta de cinco estudos, contemplando três forrageiras. Foram avaliadas 132 estimativas individuais do CMS observado (CMSObs), obtidas por meio do indicador externo Cr2O3. O CMS também foi predito por meio do software do NRC (CMSPred), que, por sua vez, foi abastecido com inputs referentes aos animais e ao ambiente de criação. Os valores de CMSPred (12,7±1,6kg/d) foram semelhantes aos de CMSObs (12,3±3,3kg/d). Foram obtidas as seguintes estimativas da avaliação do poder preditivo do modelo: viés médio (-0,419kg/d), coeficiente de determinação (0,029), coeficiente de correlação (0,17; P=,051), quadrado médio do erro de predição (11,844±20,034), fator de eficiência do modelo (-0,081), coeficiente de determinação do modelo (4,1032) e fator de correção do viés (0,767). A comparação entre CMSObs e CMSPred permitiu identificar a tendência de superestimação das predições se considerado o ajuste por meio de regressão robusta para o modelo linear simples sem intercepto. Nas condições avaliadas, o modelo produz predições de CMS com satisfatória exatidão, porém com baixa precisão.(AU)

The aim of the present study was to evaluate the predictive power of estimating the dry matter intake (DMI) of crossbred cows on tropical pastures by the National Research Council (NRC) equation for dairy cattle. A joint analysis of five studies covering three forages was performed in which 132 individual estimates of observed DMI obtained through Cr2O3 as a marker. DMI was also predicted from the NRC (DMIPred) software with inputs concerning animals and breeding environment of the studies used. Predicted DMIPred average values (12.7±1.6kg/d) were similar to the observed DMIObs ones (12.3±3.3kg/d). We obtained the following estimates of the evaluation of the predictive power of the model: average bias (- 0.419kg/d), coefficient of determination (0.029), Person's correlation coefficient (0.17, P= 0.051), mean square error of prediction (11,844±20,034), model efficiency factor (- 0.081), coefficient of determination (4.1032), and bias correction factor (0.767). The comparison between DMIObs and DMIPred values allowed the identification of the overestimating tendency of the predictions demonstrated by the robust regression fit of the simple linear no intercept model. Nevertheless, the model yields predictions with satisfactory accuracy, but with low precision.(AU)

Avaliação do modelo NRC para predição do consumo de matéria seca por vacas leiteiras manejadas em pastos tropicais / NCR Model Evaluation for prediction of dry matter intake by dairy cows managed in tropical pastures

Objetivou-se avaliar o poder preditivo do modelo do National Research Council (NRC) para gado leiteiro em estimar o consumo de matéria seca (CMS) por vacas mestiças, em pastagens tropicais. Foi efetuada uma análise conjunta de cinco estudos, contemplando três forrageiras. Foram avaliadas 132 estimativas individuais do CMS observado (CMSObs), obtidas por meio do indicador externo Cr2O3. O CMS também foi predito por meio do software do NRC (CMSPred), que, por sua vez, foi abastecido com inputs referentes aos animais e ao ambiente de criação. Os valores de CMSPred (12,7±1,6kg/d) foram semelhantes aos de CMSObs (12,3±3,3kg/d). Foram obtidas as seguintes estimativas da avaliação do poder preditivo do modelo: viés médio (-0,419kg/d), coeficiente de determinação (0,029), coeficiente de correlação (0,17; P=,051), quadrado médio do erro de predição (11,844±20,034), fator de eficiência do modelo (-0,081), coeficiente de determinação do modelo (4,1032) e fator de correção do viés (0,767). A comparação entre CMSObs e CMSPred permitiu identificar a tendência de superestimação das predições se considerado o ajuste por meio de regressão robusta para o modelo linear simples sem intercepto. Nas condições avaliadas, o modelo produz predições de CMS com satisfatória exatidão, porém com baixa precisão.(AU)

The aim of the present study was to evaluate the predictive power of estimating the dry matter intake (DMI) of crossbred cows on tropical pastures by the National Research Council (NRC) equation for dairy cattle. A joint analysis of five studies covering three forages was performed in which 132 individual estimates of observed DMI obtained through Cr2O3 as a marker. DMI was also predicted from the NRC (DMIPred) software with inputs concerning animals and breeding environment of the studies used. Predicted DMIPred average values (12.7±1.6kg/d) were similar to the observed DMIObs ones (12.3±3.3kg/d). We obtained the following estimates of the evaluation of the predictive power of the model: average bias (- 0.419kg/d), coefficient of determination (0.029), Person's correlation coefficient (0.17, P= 0.051), mean square error of prediction (11,844±20,034), model efficiency factor (- 0.081), coefficient of determination (4.1032), and bias correction factor (0.767). The comparison between DMIObs and DMIPred values allowed the identification of the overestimating tendency of the predictions demonstrated by the robust regression fit of the simple linear no intercept model. Nevertheless, the model yields predictions with satisfactory accuracy, but with low precision.(AU)

The Effects of Two Different Cleaning and Disinfection Programs on Broiler Performance and Microbiological Status of Broiler Houses

The objective of this study was to investigate the influence of two cleaning and disinfection programs on broiler performance and on the microbiological status of the facilities. This trial was an observational study of comparative character. Two experiments were conducted, with 960 birds each. Both experiments were carried out in a positive-pressure broiler house. Broilers were distributed in pens equipped with a bell drinker and a tube feeder each. In the first experiment, new wood shavings were used as litter material, and in the second, reused wood shavings were used. Two treatments with16 replicates of 30 birds each were evaluated. The regular treatment consisted of dry and wet organic matter removal, followed by washing. The European treatment consisted of dry organic matter removal, humidification, washing with water under high pressure, detergent application, rinsing, and application of two combined disinfectants: glutaraldehyde 250g/L + formaldehyde 185g/L; p-chlor-m-cresol 210 g/L. Biosecurity measures were adopted during daily management tasks to prevent cross contamination between treatments. The effectiveness of the treatments was evaluated by microbiological analysis performed before and after treatment applications, as well as by broiler performance results. Live performance results were similar between both treatments when broilers were reared on new litter. When reused word-shavings were used as litter material, the European treatment promoted better broiler performance. The European treatment was more effective than the regular treatment in reducing total microbial counts in the facilities when reused wood shavings were used as litter material, and positively influenced broiler performance.

The Effects of Two Different Cleaning and Disinfection Programs on Broiler Performance and Microbiological Status of Broiler Houses

The objective of this study was to investigate the influence of two cleaning and disinfection programs on broiler performance and on the microbiological status of the facilities. This trial was an observational study of comparative character. Two experiments were conducted, with 960 birds each. Both experiments were carried out in a positive-pressure broiler house. Broilers were distributed in pens equipped with a bell drinker and a tube feeder each. In the first experiment, new wood shavings were used as litter material, and in the second, reused wood shavings were used. Two treatments with16 replicates of 30 birds each were evaluated. The regular treatment consisted of dry and wet organic matter removal, followed by washing. The European treatment consisted of dry organic matter removal, humidification, washing with water under high pressure, detergent application, rinsing, and application of two combined disinfectants: glutaraldehyde 250g/L + formaldehyde 185g/L; p-chlor-m-cresol 210 g/L. Biosecurity measures were adopted during daily management tasks to prevent cross contamination between treatments. The effectiveness of the treatments was evaluated by microbiological analysis performed before and after treatment applications, as well as by broiler performance results. Live performance results were similar between both treatments when broilers were reared on new litter. When reused word-shavings were used as litter material, the European treatment promoted better broiler performance. The European treatment was more effective than the regular treatment in reducing total microbial counts in the facilities when reused wood shavings were used as litter material, and positively influenced broiler performance.(AU)