An updated Axiom buffalo genotyping array map and mapping of cattle quantitative trait loci to the new water buffalo reference genome assembly.

The objectives of this study were to provide the buffalo research community with an updated SNP map for the Axiom Buffalo Genotyping (ABG) array with genomic positions for SNP currently unmapped and to map all cattle QTL from the CattleQTLdb onto the buffalo reference assembly. To update the ABG array map, all SNP probe sequences from the ABG array were re-aligned against the UOA_WB_1 assembly. With the new map, the number of mapped markers increased by approximately 10% and went from 106 778 to 116 708, which reduced the average marker spacing by approximately 2 kb. A comparison of results between signatures of autozygosity study using the ABG and the new map showed that, when the additional markers were used there was an increase in the autozygosity peaks and additional peaks in BBU5 and BBU11 could be identified. After sequence alignment and quality control, 64 650 (UMD3.1) and 76 530 (ARS_UCD1.2) cattle QTL were mapped onto the buffalo genome. The mapping of the bovine QTL database onto the buffalo genome should be useful for genome-wide association studies in buffalo and, given the high homology between the two species, the positions of cattle QTL on the buffalo genome can serve as a stepping stone towards a water buffalo QTL database.

Uso da somatotropina recombinante bovina em búfalas leiteiras II: metabolismos energético e mineral / Use of recombinant bovine somatotropin (rbST) in dairy buffaloes II: energy and mineral metabolism

Avaliou-se a influência da somatotropina recombinante bovina (rbST) sobre os metabolismos energético e mineral de búfalas entre 63e 154 dias em lactação. Foram utilizadas 22 búfalas, distribuídas em dois grupos experimentais: grupo rbST - aplicação de 500mg de rbST a cada 14 dias; grupo Controle - sem aplicação de rbST. A cada sete dias, foram coletadas amostras de sangue para a determinação do perfil bioquímico e mensuraram-se a produção de leite e o escore de condição corporal dos animais. As médias dos parâmetros estudados para os grupos rbST e Controle foram, respectivamente: produção de leite (PL): 6,44kg vs. 6,68kg; escore de condição corporal-ECC (1-5): 3,51 vs. 3,57; glicose: 70,58 vs. 64,81mg/dL (P = 0,0003); colesterol: 132,38 vs. 133,40mg/dL; triglicérides: 29,18 vs. 28,32mg/dL; proteína total: 8,57 vs. 8,75g/dL; albumina: 3,47 vs. 3,60g/dL; ureia: 32,46 vs. 33,86mg/dL; creatinina: 1,27 vs. 1,39mg/dL; cálcio:10,25 vs. 10,73mg/dL; fósforo:5,76 vs. 5,62mg/dL; e magnésio:3,70 vs. 3,70mg/dL. O uso de 500mg de rbSTinfluenciou o metabolismo da glicose, porém não modificou a PL, o ECC e os níveis dos demais parâmetros metabólicos estudados.(AU)

The aim was to evaluate the influence of recombinant bovine somatotropin (rbST) on the energy and mineral metabolism of buffaloes between 63 - 154 days in milk. Twenty-two buffaloes distributed in two experimental groups were used: Group rbST (n= 11) - application of 500mg of rbST every 14 days; Control Group (n= 11) - no rbST. Every seven days, blood samples were taken to determine the biochemical profile, and milk production and body condition score were measured. The averages of the variables for rbST and Control groups were, respectively: milk yield (MY) - 6.44kg vs. 6.68kg; body condition score (BCS) - 3.51 vs 3.57 (1-5); glucose - 70.58 vs. 64.81mg/dL (P = 0.0003); cholesterol - 132.38 vs. 133.40mg/dL; triglycerides -29.18 vs. 28.32mg/dL; total protein - 8.57 vs. 8.75g/dL; albumin - 3.47 vs 3.60g/dL; urea - 32.46 vs 33.86mg/dL; creatinine - 1.27 vs 1.39mg/dL; calcium - 10.25 vs. 10.73mg/dL; phosphorus - 5.76 vs 5.62mg/dL; and magnesium - 3.70 vs 3.70mg/dL. Use of 500mg rbST influenced glucose metabolism, but did not modify the MY, BCS and the levels of the other metabolic parameters studied.(AU)

Uso da somatotropina recombinante bovina em búfalas leiteiras II: metabolismos energético e mineral / Use of recombinant bovine somatotropin (rbST) in dairy buffaloes II: energy and mineral metabolism

Avaliou-se a influência da somatotropina recombinante bovina (rbST) sobre os metabolismos energético e mineral de búfalas entre 63e 154 dias em lactação. Foram utilizadas 22 búfalas, distribuídas em dois grupos experimentais: grupo rbST - aplicação de 500mg de rbST a cada 14 dias; grupo Controle - sem aplicação de rbST. A cada sete dias, foram coletadas amostras de sangue para a determinação do perfil bioquímico e mensuraram-se a produção de leite e o escore de condição corporal dos animais. As médias dos parâmetros estudados para os grupos rbST e Controle foram, respectivamente: produção de leite (PL): 6,44kg vs. 6,68kg; escore de condição corporal-ECC (1-5): 3,51 vs. 3,57; glicose: 70,58 vs. 64,81mg/dL (P = 0,0003); colesterol: 132,38 vs. 133,40mg/dL; triglicérides: 29,18 vs. 28,32mg/dL; proteína total: 8,57 vs. 8,75g/dL; albumina: 3,47 vs. 3,60g/dL; ureia: 32,46 vs. 33,86mg/dL; creatinina: 1,27 vs. 1,39mg/dL; cálcio:10,25 vs. 10,73mg/dL; fósforo:5,76 vs. 5,62mg/dL; e magnésio:3,70 vs. 3,70mg/dL. O uso de 500mg de rbSTinfluenciou o metabolismo da glicose, porém não modificou a PL, o ECC e os níveis dos demais parâmetros metabólicos estudados.(AU)

The aim was to evaluate the influence of recombinant bovine somatotropin (rbST) on the energy and mineral metabolism of buffaloes between 63 - 154 days in milk. Twenty-two buffaloes distributed in two experimental groups were used: Group rbST (n= 11) - application of 500mg of rbST every 14 days; Control Group (n= 11) - no rbST. Every seven days, blood samples were taken to determine the biochemical profile, and milk production and body condition score were measured. The averages of the variables for rbST and Control groups were, respectively: milk yield (MY) - 6.44kg vs. 6.68kg; body condition score (BCS) - 3.51 vs 3.57 (1-5); glucose - 70.58 vs. 64.81mg/dL (P = 0.0003); cholesterol - 132.38 vs. 133.40mg/dL; triglycerides -29.18 vs. 28.32mg/dL; total protein - 8.57 vs. 8.75g/dL; albumin - 3.47 vs 3.60g/dL; urea - 32.46 vs 33.86mg/dL; creatinine - 1.27 vs 1.39mg/dL; calcium - 10.25 vs. 10.73mg/dL; phosphorus - 5.76 vs 5.62mg/dL; and magnesium - 3.70 vs 3.70mg/dL. Use of 500mg rbST influenced glucose metabolism, but did not modify the MY, BCS and the levels of the other metabolic parameters studied.(AU)

Occurrence of serological reactions for serogroup Sejroe (CTG and Prajtino) in female buffalo in the state of Pernambuco, Brazil

ABSTRACT The objective of this study was to evaluate the occurrence of anti-Leptospira spp. antibodies in female buffalo in the state of Pernambuco. A total of 123 female buffalo blood samples were collected from five properties distributed in the state of Pernambuco. The microscopic agglutination test was used to study anti-Leptospira spp. antibodies. The occurrence of anti-Leptospira spp. antibodies was 28.5% (35/123; CI 20.7-37.3%) and on different properties, the occurrence ranged from 28.6% to 80.0%, with 100% of the properties showing animals with positive results. The serovars of the serogroup Sejroe with a higher incidence were Hardjoprajtino (CTG strain, 49.1%) and Hardjo (Prajtino genotype, 43.2%), followed by serogroup Grippotyphosa with the Grippotyphosa serovar (3.9%), serogroup Pomona with the Pomona serovar (1.9%), and the Icterohaemorrhagiae serovar Copenhageni (1.9%). This was the first record of the occurrence of anti-Lepstospira spp. antibodies in female buffalo in the state of Pernambuco. Control measures are necessary to prevent health and economic losses, given that the agent involved affects animal reproduction, triggering drops in conception rates or even clinical cases of abortion.

Occurrence of serological reactions for serogroup Sejroe (CTG and Prajtino) in female buffalo in the state of Pernambuco, Brazil.

The objective of this study was to evaluate the occurrence of anti-Leptospira spp. antibodies in female buffalo in the state of Pernambuco. A total of 123 female buffalo blood samples were collected from five properties distributed in the state of Pernambuco. The microscopic agglutination test was used to study anti-Leptospira spp. antibodies. The occurrence of anti-Leptospira spp. antibodies was 28.5% (35/123; CI 20.7-37.3%) and on different properties, the occurrence ranged from 28.6% to 80.0%, with 100% of the properties showing animals with positive results. The serovars of the serogroup Sejroe with a higher incidence were Hardjoprajtino (CTG strain, 49.1%) and Hardjo (Prajtino genotype, 43.2%), followed by serogroup Grippotyphosa with the Grippotyphosa serovar (3.9%), serogroup Pomona with the Pomona serovar (1.9%), and the Icterohaemorrhagiae serovar Copenhageni (1.9%). This was the first record of the occurrence of anti-Lepstospira spp. antibodies in female buffalo in the state of Pernambuco. Control measures are necessary to prevent health and economic losses, given that the agent involved affects animal reproduction, triggering drops in conception rates or even clinical cases of abortion.

Brazilian ground beef authentication by multiplex polymerase chain reaction / Autenticação de carne moída brasileira através de uma reação em cadeia da polimerase multiplex

The aim of the present study was to assess the efficacy ofmultiplex PCR in detecting the adulterationof commercially available ground beefvia addition and/orsubstitution ofground buffalo meat. Experimentally adulterated ground beefsamples were prepared in triplicate, and dilutions of DNA from Bos taurus and Bubalusbubalis were prepared to determine the detection limit of the method. Concurrently, 91 ground meatsamples sold as ground beef were collected from differentstores in northern Brazil andanalyzed bymultiplex PCR. Buffalo DNA was detected in 17.5% of the collected ground meat samples.Our results showed that multiplex PCR is an efficient method for detectingthe incorporation of groundbuffalo meatatpercentages ranging from 10 to 100% and the incorporation of beef at percentages ranging from0.1 to 100% intoground meat samples.(AU)

O objetivo do presente estudo foi avaliar a eficácia da PCR multiplex na detecção da adulteração por adição e/ou substituição de carne bubalina em carne moída bovina, comercialmente disponível. A fim de determinar o limite de detecção da técnica, carnes moídas bovinas fraudadas intencionalmente foram fabricadas em triplicata e, adicionalmente, concentrações conhecidas de DNA das espécies Bostaurus e Bubalusbubalis foram diluídos. Ao mesmo tempo, 91 amostras de carne moída comercializadas como sendo de origem bovina foram coletadas em diferentes comércios na região Norte do Brasil e a PCR multiplex proposta foi realizada. Os resultados demonstraram que o DNA bubalino foi detectado em 17,5% das amostras de carne moída coletadas. Concluiu-se que a PCR multiplex é uma técnica eficaz, capaz de detectar a incorporação de carne moída bubalina em percentagens que variaram de 10 a 100%, e a incorporação de carne bovina em percentagens que variaram de 0,1 a 100% em amostras de carne moída.(AU)

Brazilian ground beef authentication by multiplex polymerase chain reaction / Autenticação de carne moída brasileira através de uma reação em cadeia da polimerase multiplex

ABSTRACT: The aim of the present study was to assess the efficacy ofmultiplex PCR in detecting the adulterationof commercially available ground beefvia addition and/orsubstitution ofground buffalo meat. Experimentally adulterated ground beefsamples were prepared in triplicate, and dilutions of DNA from Bos taurus and Bubalusbubalis were prepared to determine the detection limit of the method. Concurrently, 91 ground meatsamples sold as "ground beef" were collected from differentstores in northern Brazil andanalyzed bymultiplex PCR. Buffalo DNA was detected in 17.5% of the collected ground meat samples.Our results showed that multiplex PCR is an efficient method for detectingthe incorporation of groundbuffalo meatatpercentages ranging from 10 to 100% and the incorporation of beef at percentages ranging from0.1 to 100% intoground meat samples.

RESUMO: O objetivo do presente estudo foi avaliar a eficácia da PCR multiplex na detecção da adulteração por adição e/ou substituição de carne bubalina em carne moída bovina, comercialmente disponível. A fim de determinar o limite de detecção da técnica, carnes moídas bovinas fraudadas intencionalmente foram fabricadas em triplicata e, adicionalmente, concentrações conhecidas de DNA das espécies Bostaurus e Bubalusbubalis foram diluídos. Ao mesmo tempo, 91 amostras de carne moída comercializadas como sendo de origem bovina foram coletadas em diferentes comércios na região Norte do Brasil e a PCR multiplex proposta foi realizada. Os resultados demonstraram que o DNA bubalino foi detectado em 17,5% das amostras de carne moída coletadas. Concluiu-se que a PCR multiplex é uma técnica eficaz, capaz de detectar a incorporação de carne moída bubalina em percentagens que variaram de 10 a 100%, e a incorporação de carne bovina em percentagens que variaram de 0,1 a 100% em amostras de carne moída.

Identificação espécie-específica de carnes e produtos cárneos de origem bubalina e bovina pela técnica de PCR-RFLP / Species-specific identification of red-meat and meat derivative products with the PCR-RFLP technique

Para avaliar a viabilidade da metodologia da Reação em Cadeia da Polimerase associada com o Polimorfismo de Fragmentos de DNA (PCR-RFLP) na identificação de fraude intencional e contaminação acidental em produtos cárneos de origem bubalina, in natura e processados, foram testadas amostras puras e amostras de carnes com misturas controladas, produzidas em laboratório, com adição de 1%, 5%, 10% e 50% de carne bovina em carne de búfalo, homogeneizada crua e em amostras autoclavada. Foram comparados, ainda, diferentes métodos de extração, usando um kit comercial e a técnica clássica, utilizando fenol/clorofórmio. O resultado estatístico foi obtido por tabela de contingência, analisada pelo teste do qui-quadrado (2) e do exato de Fisher. A especificidade encontrada foi altamente significativa (P<0,0001). Observou-se também sensibilidade altamente significativa nas diluições a partir de 10% (P<0,0001). A técnica tem alta especificidade e sensibilidade para detectar até mesmo contaminação de 1%, mas a repetibilidade desse resultado impede a aplicação oficial desse método para a inspeção de contaminação acidental, sendo recomendada somente para inspeção de fraude a partir de 10% de substituição. Em carnes autoclavadas, a eficácia do teste é menor. A técnica pode ser empregada para certificação de produto específico (selo de identidade de espécie).(AU)

The present study aimed at evaluate the viability of PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) for identification of fraud and/or accidental contamination in buffalo meat - fresh and processed. Pure, autoclaved and controlled fraud samples, produced in the laboratory with the addition of 1, 5, 10 and 50% of beef in raw homogenized buffalo meat samples, were tested. Furthermore, different extraction methods, using a commercial kit and classical technique using phenol-chloroform, were compared. The statistical result was obtained by contingency table analyzed by chi-square and the Fisher exact test. The specificity was highly significant (p <0.0001), and the sensitivity was highly significant in dilutions from 10% (p <0.0001). Despite its accuracy and precision, capable to detect a contamination level of 1%, PCR-RFLP technique is not recommended for inspection in cases of accidental contamination. This is due to the need of test repetition in levels of contamination lower than 10%. The efficiency of this test is lower to autoclaved meat. The PCR-RFPL technique can be used for certification of food made with specific species (species identification certification stamp).(AU)

Identificação espécie-específica de carnes e produtos cárneos de origem bubalina e bovina pela técnica de PCR-RFLP / Species-specific identification of red-meat and meat derivative products with the PCR-RFLP technique

Para avaliar a viabilidade da metodologia da Reação em Cadeia da Polimerase associada com o Polimorfismo de Fragmentos de DNA (PCR-RFLP) na identificação de fraude intencional e contaminação acidental em produtos cárneos de origem bubalina, in natura e processados, foram testadas amostras puras e amostras de carnes com misturas controladas, produzidas em laboratório, com adição de 1%, 5%, 10% e 50% de carne bovina em carne de búfalo, homogeneizada crua e em amostras autoclavada. Foram comparados, ainda, diferentes métodos de extração, usando um kit comercial e a técnica clássica, utilizando fenol/clorofórmio. O resultado estatístico foi obtido por tabela de contingência, analisada pelo teste do qui-quadrado (χ2) e do exato de Fisher. A especificidade encontrada foi altamente significativa (P<0,0001). Observou-se também sensibilidade altamente significativa nas diluições a partir de 10% (P<0,0001). A técnica tem alta especificidade e sensibilidade para detectar até mesmo contaminação de 1%, mas a repetibilidade desse resultado impede a aplicação oficial desse método para a inspeção de contaminação acidental, sendo recomendada somente para inspeção de fraude a partir de 10% de substituição. Em carnes autoclavadas, a eficácia do teste é menor. A técnica pode ser empregada para certificação de produto específico (selo de identidade de espécie).

The present study aimed at evaluate the viability of PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) for identification of fraud and/or accidental contamination in buffalo meat - fresh and processed. Pure, autoclaved and controlled fraud samples, produced in the laboratory with the addition of 1, 5, 10 and 50% of beef in raw homogenized buffalo meat samples, were tested. Furthermore, different extraction methods, using a commercial kit and classical technique using phenol-chloroform, were compared. The statistical result was obtained by contingency table analyzed by chi-square and the Fisher exact test. The specificity was highly significant (p <0.0001), and the sensitivity was highly significant in dilutions from 10% (p <0.0001). Despite its accuracy and precision, capable to detect a contamination level of 1%, PCR-RFLP technique is not recommended for inspection in cases of accidental contamination. This is due to the need of test repetition in levels of contamination lower than 10%. The efficiency of this test is lower to autoclaved meat. The PCR-RFPL technique can be used for certification of food made with specific species (species identification certification stamp).