Spoligotyping, genotyping, and spatial distribution of Mycobacterium bovis in cattle in the state of Bahia, Brazil / Genotipagem por spoligotyping e distribuição espacial de Mycobacterium bovis em bovinos no estado da Bahia, Brasil

Mycobacterium bovis is responsible for bovine and buffalo tuberculosis, an important zoonotic disease with global distribution. The knowledge of the distribution and the precise identification of this disease, including advanced diagnoses such as spoligotyping, allows choosing the best strategies to fight the disease's progress. The present work aimed to investigate mycobacteria's presence, genotype their strains, and evaluate tuberculosis cases' spatial distribution from suggestive lesions in carcasses of bovine and buffalo inspected in slaughterhouses under an official inspection regime in the state of Bahia, Brazil. The study investigated 453,417 animals. Among these, 31 (0.007%) from 17 municipalities were suspected of tuberculosis. Among the culture medium growth, 95% of these were categorized as alcohol-acid resistant bacilli (BAAR). All isolates were subjected to spoligotyping and 95% were confirmed as M. bovis (SB0120, SB0121, SB0852, SB0828, SB0295, SB0881, SB1648, SB6119, SB0140, SB1055). The strain SB0120 was the most prevalent, and this profile has been described in cases of human tuberculosis by M. bovis, highlighting the zoonotic potential of this profile. This study also identified strains never reported in Bahia, highlighting a distinctive pattern from other parts of Brazil, besides mixed infections. Besides, to identify strains never before described in the state, highlighting a distinctive pattern in Brazil (SB6119 and SB0852, respectively). An unpublished profile was identified and inserted in the international database (Mbovis.org), named SB2715.(AU)

O Mycobacterium bovis é o responsável pela tuberculose bovina e bubalina, doença zoonótica importante e com distribuição global. O conhecimento da distribuição e a identificação precisa dessa enfermidade, incluindo diagnósticos mais avançados como o spoligotyping, permite escolher as melhores estratégias de combate ao avanço da doença. O presente trabalho objetivou investigar a presença de micobactérias, genotipar suas estirpes e avaliar a distribuição espacial dos casos de tuberculose a partir de lesões sugestivas nas carcaças de bovinos e bubalinos inspecionadas em frigoríficos sob regime de inspeção oficial no estado da Bahia. Foram investigados 453.417 animais dentre os quais 31 (0,007%) foram suspeitos de doença e provenientes de 17 municípios. Após o crescimento em meio de cultura, 95% foram categorizados como bacilos álcool-ácido resistentes (BAAR). Todos os isolados foram submetidos à spoligotyping e 95% foram confirmados M. bovis (SB0120, SB0121, SB0852, SB0828, SB0295, SB0881, SB1648, SB6119, SB0140, SB1055). A cepa SB0120 foi a mais prevalente e este perfil vem sendo descrito na literatura com casos de tuberculose humana por M. bovis ressaltando o potencial zoonótico deste perfil. Este estudo também identificou cepas nunca relatadas no estado da Bahia, destacando um padrão distinto de outras partes do Brasil, além da existência de infecções mistas. Permitiu ainda relatar linhagens nunca antes descritas no estado com destaque para um padrão novo no Brasil (SB6119 e SB0852 respectivamente). Um perfil inédito identificado foi identificado e inserido no banco de dados internacional (Mbovis.org), nomeado SB2715.(AU)

Productive characteristics in dairy buffalo (Bubalus bubalis) in the Eastern Amazon / Características produtivas em búfalos leiteiros (Bubalus bubalis) na Amazônia

The objective of this study was to determine factors affecting phenotypic characteristics related to milk production and reproductive efficiency of dairy buffalo. Phenotypic records of 2,459 Murrah, Mediterranean and crossbred buffaloes from the Embrapa Eastern Amazônia herd, from 1969 to 2013 were used. Traits included: total milk production during lactation (TMP), milk production corrected for 305 days (P305); cow weight at calving (CW), fat milk percentage (F), age at first calving (AFC), calving interval (CI), calf birth weight (BW), as well as lactation (LP) and service periods (SP). Most traits were affected by genetic group as well as year and season or interaction between them. Genetic group affected F, CI, SP and CW. Very young and older cows showed poorer production traits. There was interaction between season and year for F, CI and CW. Correlations traits were generally medium to low. The principal component analysis showed that higher fat milk percentage was associated with higher percentage of Murrah breed, longer lactation period and older cows. Phenotypic characteristics in buffaloes reared in the Brazilian Amazon were significantly affected by genetic and management factors. This information can be used to help build sustainable production systems for the region and guide farmer decisions.(AU)

O objetivo deste estudo foi determinar os fatores que afetam as características fenotípicas relacionadas à produção de leite e a eficiência reprodutiva de búfalas leiteiras. Foram utilizados os registros fenotípicos de 2.459 búfalas das raças Murrah, Mediterrâneo e seus cruzamentos do rebanho da Embrapa Amazônia Oriental, de 1969 a 2013. As características incluídas foram: produção total de leite durante a lactação (PTL), produção de leite corrigida para 305 dias (P305), peso da búfala ao parto (PV), percentual de gordura do leite (G), idade ao primeiro parto (IPP), intervalo entre partos (IP), peso do bezerro ao nascimento (PN), assim como os períodos de lactação (PL) e de serviço (PS). A maioria das características foi afetada pelo grupo genético, bem como pelo ano e pela estação ou pela interação entre eles. O grupo genético afetou o G, IP, PS e PV. Búfalas muito jovens e mais velhas mostraram características de produção mais baixas. Houve interação entre estação e o ano para G, IP e PV. As correlações entre as características foram geralmente de médias a baixas. A análise de componentes principais mostrou que a maior porcentagem de gordura no leite foi associada com a maior porcentagem de sangue Murrah, com o maior período de lactação e com búfalas mais velhas. As características fenotípicas das búfalas criadas na Amazônia brasileira foram significativamente afetadas por fatores genéticos e de manejo. Essas informações podem ser usadas para ajudar a construir sistemas de produção sustentáveis para a região e orientar as decisões dos produtores.(AU)

cDNA Cloning of a Bovine Insulin-like growth factor-1 from Egyptian Buffalos and Expression of its Recombinant Protein in Escherichia coli / Clonagem de cDNA de um fator-1 de crescimento semelhante à insulina bovina de búfalos egípcios e expressão de sua proteína recombinante em Escherichia coli

Insulin-like growth factor-1 (IGF-1) is regarded as a crucial clinically significant therapeutic agent against several pathological conditions. Recently, recombinant DNA (rDNA) technology has enabled the production of many drugs of rDNA-origin including IGF-1. Securing a readily available supply of IGF-1 is invaluable to clinical research and biotechnological domains. In this work, the cloning of a full-length bovine IGF-1 cDNA and the successful expression of its cognate recombinant IGF-1 protein is reported. Single-strand cDNA was prepared from liver tissues, through the specific reverse transcription (RT) of IGF-1 mRNA. Subsequently, a PCR amplicon of ~543bp was successfully amplified. Recombinant pTARGET™ vector harboring IGF-1 insert was successfully cloned into competent E. coli JM109 cells. SDS-PAGE analysis revealed that the recombinant IGF-1 has been expressed at the expected size of 7.6kDa. The outcome provides a robust basis for transecting the recombinant pTARGETTM vector, harboring the IGF-1 cDNA insert, into mammalian cells. Optimal initial glucose concentration was found to be 10g/l with corresponding protein concentration of 6.2g/l. The proliferative biological activity crude recombinant IGF-1 protein was verified on HeLa cell lines. This is envisaged to facilitate large-scale production of recombinant IGF-1 protein, thereby enabling thorough investigation of its clinical and pharmaceutical effects.(AU)

O fator de crescimento semelhante à insulina-1 (IGF-1) é considerado um agente terapêutico clinicamente significativo contra várias condições patológicas. Recentemente, a tecnologia de DNA recombinante (rDNA) permitiu a produção de muitos medicamentos de origem rDNA, incluindo o IGF-1. Garantir um suprimento prontamente disponível de IGF-1 é inestimável para pesquisas clínicas e domínios biotecnológicos. Neste trabalho, relata-se a clonagem de um cDNA de IGF-1 bovino de comprimento total e a expressão bem-sucedida de sua proteína IGF-1 recombinante cognata. O cDNA de cadeia simples foi preparado a partir de tecidos do fígado, por meio da transcrição reversa específica (RT) do mRNA de IGF-1. Posteriormente, um amplificador de PCR de ~ 543pb foi amplificado com sucesso. O vetor pTARGET™ recombinante contendo a inserção de IGF-1 foi clonado com sucesso em células competentes E. coli JM109. A análise por SDS-PAGE revelou que o IGF-1 recombinante foi expresso no tamanho esperado de 7,6kDa. O resultado fornece uma base robusta para a transferência do vetor pTARGETTMTM recombinante, abrigando a inserção de cDNA de IGF-1 em células de mamíferos. Verificou-se que a concentração inicial ideal de glicose é 10g/L, com a concentração de proteína correspondente de 6,2g/L. A proteína IGF-1 recombinante bruta de atividade biológica proliferativa foi verificada nas linhas celulares HeLa. É previsto que isso facilite a produção da proteína IGF-1 recombinante em larga escala, permitindo, assim, uma investigação completa dos seus efeitos clínicos e farmacêuticos.(AU)

Genetic evaluation of milking buffaloes (Bubalus bubalis): bull ranking / Avaliação genética de búfalas leiteiras (Bubalus bubalis): classificação de touros

The objective of this study was to evaluate genetic aspects related to production and reproductive efficiency of Murrah and Mediterranean buffaloes and their crosses. A ranking of bulls from Embrapa Eastern Amazonia was also composed to guide assisted mating. Birth records of 2,322 Murrah, Mediterranean, and crossbred buffaloes from the Embrapa Eastern Amazon herd, from 1953 to 2013, as well as information on production and reproductive traits were used. Genetic analyzes were performed in the WOMBAT software using the animal model with two-trait analysis. While heritability (h2) for total milk production (TMP) and fat milk percentage (F) were generally high, for reproductive traits h2 tended to be low. Genetic correlations for TMP with the other traits were low and negative, except for TMP with calving interval (CI) and service period (SP) in the Mediterranean breed and with age at first calving (AFC) and SP in crossbred, which were positive and high. Bull 1001 had high predicted transmitting ability (PTA) for TMP, so it should transmit a greater volume of milk to his offspring, although it had a lower PTA for F. There was sufficient variability within the herd to work with genetic management for both production and reproductive efficiency.(AU)

O objetivo deste estudo foi avaliar os aspectos genéticos relacionados à produção e à eficiência reprodutiva de búfalas das raças Murrah, Mediterrâneo e suas cruzas. Uma classificação de touros da Embrapa Amazônia Oriental também foi composta para orientar os cruzamentos assistidos. Foram utilizados 2.322 registros de nascimento de búfalas das raças Murrah, Mediterrâneo e cruzadas do rebanho da Embrapa Amazônia Oriental, de 1953 a 2013, bem como características produtivas e reprodutivas. As análises genéticas foram realizadas pelo software Wombat, utilizando-se o modelo animal com análise de duas características. Enquanto a herdabilidade (h 2 ) para a produção total de leite (PTL) e para a porcentagem de gordura (G) foi alta, para as características reprodutivas a h 2 tendeu a ser baixa. As correlações genéticas da PTL com as demais características foram baixas e negativas, exceto para a PTL com intervalo entre partos (IP) e período de serviço (PS) na raça Mediterrâneo e com idade ao primeiro parto (IPP) e PS nas cruzadas, que foram positivas e altas. O touro 1001 apresentou alta capacidade de transmissão predita (CTP) para a PTL, então deve transmitir um maior volume de leite para seus descendentes, embora com um menor conteúdo transmissível de CTP para G. Portanto, existe variabilidade suficiente dentro do rebanho para trabalhar com o manejo genético para a produção e a eficiência reprodutiva.(AU)

Molecular detection of albinism gene in Brazilian buffalo herds (Bubalus bubalis) / Detecção molecular do gene do albinismo em rebanhos de búfalos (Bubalus bubalis) do Brasil

Albinism is a genetic disease characterized by deficient melanin production making affected animals more susceptible to skin problems, negatively influencing production systems of the same. In buffalo, a nonsense mutation (c.1431G>A) in the tyrosinase gene was already described, which is responsible for the oculocutaneous albinism buffalo phenotype. However, prevalence studies have never been performed for this anomaly in Brazil. Therefore, the objective of this study was to investigate this mutation in buffalo herd in Brazil. Of the 315 buffalo tested with no albinism phenotype evident, 11 (3.5%) were heterozygous for the mutation and none were mutated homozygous, showing the existence of the albinism gene in buffalo production herds and proving the importance of prevalence studies for hereditary diseases in order to prevent the dissemination of these same genes and their negative productivity consequences.(AU)

O Albinismo é uma doença genética caracterizada pela deficiência na produção de melanina, o que torna os animais afetados mais susceptíveis a problemas cutâneos e influencia negativamente a criação destes animais. A mutação nonsense (c.1431G>A) no gene da tyrosinase já foi descrita como responsável pelo albinismo oculocutâneo em búfalos, entretanto estudos prévios sobre a prevalência dessa mutação ainda não foram realizados no Brasil. Portanto, o objetivo deste estudo foi avaliar a presença desta mutação em uma população de búfalos brasileiros. Foram genotipados 315 búfalos clinicamente normais, ou seja, sem o fenótipo albino evidente. Desses, 11 (3,5%) eram heterozigotos para a mutação (N/TYR) e os demais eram homozigotos selvagens (N/N). Este resultado demonstra que o alelo mutado para o albinismo em búfalo está presente no rebanho brasileiro e aponta a importância de estudos de prevalência de enfermidades hereditárias com o objetivo de prevenir a disseminação desses alelos mutados, minimizando os prejuízos.(AU)

Pretentious genomic selection signatures in CYP19A1 gene associated with silent estrous behavior in water buffalo in Pakistan

Background: Poor reproductive efficiency of river buffalos hampers the production capabilities of animals. Buffalos are mainly considered poor breeders owing to the constrained expression of estrus behavior. Failure to display heat signs is an indication of improper functionality of signaling peptides to trigger on a series of behavioral changes, which can be detectable by breeders for timely insemination of females. This might cause an animal to be a repeat breeder. Genomic variations underlying synthesis of signaling peptides can be a useful marker to select superior animals with better reproductive efficiency. In this context, the current study was designed to analyze the CYP19A1 gene in Nili-Ravi buffalo. Results: A total of 97 animals were selected and were divided into two groups on the basis of their heat score. PCR amplification and sequencing of the amplicons were performed using the specific sets of primer, and then, sequences were analyzed for novel variants. A total of 11 polymorphic sites were identified illustrating phenotypic variation in the heat score. Most of the loci were found homologous. Single Nucleotide Polymorphisms (SNPs) were analyzed for association with silent estrus. A three-dimensional protein model was also generated to locate the position of exonic SNPs. Conclusion: This study illustrated that polymorphic sites in the CYP19A1 gene provided potential markers for selection of buffalos with better estrus behavior.

Variabilidade genética do peso ao nascer e seleção para crescimento em bubalinos do Estado do Pará, Brasil / Genetic variability of birth weight and selection for growth of water buffaloes from State of Pará, Brazil

Birth weight is a performance parameter of great zootechnical importance for both meat and dairy production, as well as for breeding animals, mainly due to its relation to survival rate at weaning, and the weight of animals throughout their developmental and growth phases. Therefore, the objective of this work was to establish heritability estimates, as well as phenotypic and genetic trends of birth weight of water buffaloes from State of Pará, Brazil. Descriptive statistics were calculated and the Shapiro-Wilk Normality test was performed with a Statistical Analysis System package. Heritability estimates were established by Bayesian inference. BW was 36.6 kg in average. The statistical model considered sex, year of birth and breed composition of the animals as fixed effects, and animal, maternal and residual as random effects. Direct heritability was platykurtic (flattened) and with higher asymmetry, presenting a bimodal distribution with the first mode close to 0.10, and the second mode close to 0.30; the maternal heritability was trimodal, with peaks very close to 0.15, and another, less evident, close to 0.20. The direct genetic trend of birth weight was negative (-0.03kg year-1) and maternal genetic trend was close to zero (0.001kg ano-1), even though the phenotypic trend had been positive (0.156 kg year-1). There is genetic variability to be addressed in a breeding program, however, very little was done as far as selection for growth of water buffaloes in the State of Pará.

O peso ao nascer constitui característica produtiva de elevada importância zootécnica, devido à sua relação com a taxa de sobrevivência à desmama e com os pesos nas demais fases de desenvolvimento do animal, quer seja para a produção de carne, leite ou em animais que se destinam à reprodução. O objetivo deste trabalho foi obter estimativas de herdabilidade e tendências fenotípicas e genéticas do peso ao nascer, em bubalinos do Estado do Pará, Brasil. Foram calculadas as estatísticas descritivas e realizado o teste de Normalidade de Shapiro-Wilk por meio do pacote estatístico Statistical Analisys System. As estimativas de herdabilidade foram obtidas por inferência Bayesiana. O peso ao nascer apresentou média de 36,6 kg. O modelo de análise considerou como fixos os efeitos de sexo, ano de nascimento e composição racial do animal e como efeitos aleatórios animal, efeito materno e residual. A distribuição da herdabilidade direta apresentou-se platicúrtica (achatada) e com maior assimetria, tendo uma distribuição bimodal com a primeira moda próxima a 0,10 e a segunda próxima a 0,30; a materna apresentou-se trimodal, com picos bem próximos a 0,15 e outro menos evidente próximo a 0,20. A tendência genética direta do peso ao nascer mostrou-se negativa (-0,03kg ano-1) e a tendência genética materna próxima à zero (0,001 kg ano-1), ainda que a tendência fenotípica tenha sido positiva (0,156kg ano-1). Existe variabilidade genética possível de ser trabalhada em um programa de melhoramento, no entanto, pouco foi feito quanto à seleção para crescimento em búfalos no Estado do Pará.

Identification of three novel myeloid cathelicidin cDNAs and their predicted peptides in buffalo (Bubalus bubalis).

Antimicrobial peptides (AMPs) are broad spectrum antibiotics, which mostly act without specific receptors. Identification of AMPs is important in the current scenario of emerging multi-drug resistant bacteria. In the present study, in an attempt to identify new AMPs, myeloid cathelicidin cDNAs were synthesized from buffalo (Bubalus bubalis) bone marrow and were amplified using specific primers. Sequence analysis of cloned cDNAs revealed three novel myeloid cathelicidins. They were named based on the number of active amino acids in the C-terminal region of their predicted peptide sequences as BuMAP-28 (having an additional Gly at position 22nd), BuMAP-29 (having an additional IIe at position 27) and BuMAP-34, compared to BMAP-27, BMAP-28 and BMAP-34 of cattle. The BuMAPs showed 93%, 95% and 87% homology respectively with that of its cattle counterpart. Predicted number of amino acids of the cDNAs was 159, 155 and 157 residues, with cationic C-terminal sequences of 28, 29 and 34, respectively, which correspond to putative antimicrobial domains. Several amino acid substitutions were observed in all the three cathelicidins. The conformation of the peptides was predicted to be alpha helical, having total net positive charge and hydrophobicity, similar to that of BMAPs in cattle. Comparative analysis of the predicted peptides suggested potential antimicrobial activity and the sequence variations detected might enable the peptides to act as effective broad spectrum antimicrobial agents.

Doenças hereditárias e defeitos congênitos diagnosticados em búfalos (Bubalus bubalis) no Brasil / Hereditary diseases and congenital defects diagnosed in water buffalo (Bubalus bubalis) in Brazil

A review on hereditary diseases and/or congenital defects diagnosed in water buffaloes in Brazil is performed. The epidemiological, clinical and pathological aspects of each disease or group of diseases are briefly described. Hereditary diseases include acantholytic mechanobullous dermatosis, arthrogryposis, myotonia, hydranencephaly, chondrodysplasia, and albinism. Congenital defects of unknown cause include megaesophagus, heart defects (patent ductus arteriosus), dermatosparaxia, and different defects of the reproductive system. The breeds most affected by genetic diseases are those from Asian Continent (Murrah and Jafarabadi), probably as a result of inbreeding in Brazilian herds due the prohibition of importation of breeding buffalo from that continent. The diagnosis of two hereditary diseases, arthrogryposis and myotonia, in Rio Grande do Sul (southern Brazil) and Pará (nothern Brazil) suggests that the undesirable genes are widespread in the buffalo population. The identification of these genes by molecular techniques associated with the buffalo breeding with correct sanitary, zootechnical, and reproductive control practices can decrease the negative effects of genetic diseases in the Brazilian buffalo herd.

É realizada uma revisão sobre as doenças hereditárias e/ou defeitos congênitos diagnosticados em búfalos no Brasil. São descritos brevemente os aspectos epidemiológicos, clínicos e patológicos de enfermidades hereditárias ou provavelmente hereditárias já observadas no Brasil, como dermatose mecanobolhosa, artrogripose, miotomia, hidranencefalia, condrodisplasia e albinismo; e dos defeitos congênitos que não tem uma causa ainda comprovada como megaesôfago, defeitos cardíacos (persistência do ducto arterioso), dermatosparaxia, defeitos no sistema reprodutivo e outros defeitos. Observou-se que as raças mais afetadas por enfermidades de natureza genética são as que têm origem no Continente Asiático (Murrah e Jafarabadi), provavelmente em consequência da consanguinidade existente nos rebanhos devido a proibição da importação de reprodutores, sêmen e embriões daquele continente. O diagnóstico de duas dessas doenças, artrogripose e miotomia hereditária no Rio Grande do Sul e no Pará, demonstra que os genes indesejáveis estão disseminados na população de búfalos no país e que a identificação desses genes por meio de técnicas moleculares associada à criação desta espécie com maior controle sanitário, reprodutivo e zootécnico pode minimizar os prejuízos decorrentes dessas enfermidades à bubalinocultura.

Characterization of PRLR and PPARGC1A genes in buffalo (Bubalus bubalis)

More than 40 million households in India depend at least partially on livestock production. Buffaloes are one of the major milk producers in India. The prolactin receptor (PRLR) gene and peroxisome proliferators activated receptor-γ coactivator 1-alpha (PPARGC1A) gene are reportedly associated with milk protein and milk fat yields in Bos taurus. In this study, we sequenced the PRLR and PPARGC1A genes in the water buffalo Bubalus bubalis. The PRLR and PPARGC1A genes coded for 581 and 819 amino acids, respectively. The B. bubalis PRLR gene differed from the corresponding Bos taurus at 21 positions and four differences with an additional arginine at position 620 in the PPARGC1A gene were found in the amino acid sequence. All of the changes were confirmed by cDNA sequencing. Twelve buffalo-specific single nucleotide polymorphisms (SNPs) were identified in both genes, with five of them being non-synonymous.

Variabilidade genética de búfalos em rebanho-núcleo com base na análise de pedigree / Genetic variability in water buffalo from nucleous herd by pedigree analysis

Parâmetros baseados na probabilidade de origem do gene foram usados para descrever a variabilidade em uma população de búfalos da Embrapa Amazônia Oriental. A magnitude dos resultados foi de média a baixa (por volta de 20 animais), sugerindo que poucos fundadores contribuiriam para a formação da população. Dentre os 20 ancestrais que mais aportam genes aos machos - representando ao todo 71 por cento dos alelos -, 39 por cento, 26 por cento e 5 por cento, respectivamente, são as contribuições marginais das raças Murrah e Mediterrâneo e seus mestiços. Para as fêmeas, em que os 20 ancestrais aportam 67,5 por cento dos genes, 42 por cento e 26 por cento, respectivamente, são as contribuições marginais das raças Murrah e Mediterrâneo.

Parameters based on the probability of gene origin were used to describe genetic variability in a buffalo population from the Embrapa Amazônia Oriental, Belém, Pará, Brazil. The parameters generated medium to low values (around 20 animals) and suggested low founder representativeness. From the 20 ancestors that gave more genes to males (with 71 percent of alleles), genetic contributions were 39 percent, 26 percent, and 5 percent, respectively, for Murrah, Mediterraneo, and crossbreds. For females, these values were 42 percent and 26 percent for Murrah and Mediterraneo breeds.

Estimação de parâmetros genéticos para características produtivas em búfalos na Amazônia Oriental / Estimation of genetic parameters of dairy buffaloes productive characteristics Eastern Amazon

Dados de 1.182 registros de produção de fêmeas bubalinas da raça Murrah e seus mestiços, parindo no período de 1967 a 2005, foram utilizados para estimação de parâmetros genéticos utilizando-se o método de máxima verossimilhança restrita. O modelo animal utilizado para estimação de componentes de variância incluiu os efeitos fixos de rebanho, ano e época de parto, ordem de parto e duração da lactação e os efeitos aleatórios do animal, e ambiente permanente e temporário. As estimativas de herdabilidade foram 0,25, 0,18, 0,08 e 0,09, para produção de leite, produção de gordura, duração da lactação e produção de leite por dia de intervalo de parto, respectivamente. As estimativas de repetibilidade foram 0,33, 0,29 e 0,10 para produção de leite, produção de gordura e duração da lactação, respectivamente. As correlações genéticas entre produções de leite e gordura, produção de leite com duração da lactação, produção de leite com produção de leite por dia de intervalo de partos, produção da gordura com duração da lactação, produção de gordura com produção de leite por dia de intervalo de partos e duração da lactação com produção de leite por dia de intervalo de partos foram 0,93; 0,76; 0,99; 0,89; 0,87 e -0,27, respectivamente. Os resultados demonstram que ganhos genéticos podem ser obtidos pela seleção das produções de leite e gordura.

Data from 1,182 records of Murrah females buffaloes and their crossbreds, calving from 1967 to 2005, were used to estimate genetic parameters by maximum restricted likelihood method. An animal model used to estimate variance components, included the fixed effects of herds, year and season of calving, order of parity, and length of lactation as covariables; and random effects of animal, permanent, and temporary environment. The estimate of heritability to milk yield, fat yield, length of lactation, and milk yield per day of calving interval were 0.25, 0.18, 0.08, and 0.09 respectively. Estimate of repeatability for milk yield, fat yield, and length of lactation were 0.33, 0.29, and 0.10, respectively. The genetic correlations between milk yield and fat yield, milk yield and length of lactation, milk yield and milk yield per day of calving interval, fat yield and length of lactation, fat yield and milk yield per day of calving interval, and length of lactation and milk yield per day of calving interval were 0.93, 0.76, 0.99, 0.89, 0.87, and -0.27, respectively. These results showed that genetic gains may be obtained by selecting to milk and fat yields.

Growth Curves and Genetic Parameters in ColombianBuffaloes (Bubalus bubalis Artiodactyla, Bovidae) / Curvas de crecimiento y parámetros genéticos en búfalos (Bubalus bubalisArtiodactyla, Bovidae) en Colombia / Curvas de crescimento e parâmetros genéticos em búfalos (Bubalus bubalisArtiodactyla, Bovidae) na Colômbia

Non-linear mathematical functions proposed by Brody, Gompertz, Richards, Bertalanffy and Verhulstwere compared in several buffalo production systems in Colombia. Herds were located in three provinces:Antioquia, Caldas, and Cordoba. Growth was better described by the curves proposed by Brody andGompertz. Using the datasets from herds from Caldas, heritabilities for traits such as weaning weight(WW), weight and maturity at one year of age (WY and MY, respectively), age at 50% and 75% of maturity(A50% and A75%, respectively), adult weight (β0), and other characteristics, were also estimated. Directand maternal heritabilities for WW were 0.19 and 0.12, respectively. Direct heritabilities for WY, MY,A50%, A75% and β0 were 0.39, 0.15, 0.09, 0.20 and 0.09, respectively. The genetic correlation for β0 andWY was -0.47, indicating that selection for heavy weight at one year of age will lead to lower weight at adultage. These data suggest that selection based on maturity traits can generate changes in characteristics ofeconomic importance in beef–type buffalo farms.

Funciones matemáticas no lineales propuestas por Brody, Gompertz, Richards, Bertalanffy y Verhulst fueron utilizadas para describir el crecimiento en búfalos provenientes de tres regiones de Colombia (Antioquia,Caldas y Córdoba). Las funciones que mejor describieron el crecimiento fueron Brody y Gompertz. Con labase de datos de la región de Caldas, fueron estimadas las heredabilidades para las características peso aldestete (PD), peso y madurez al año de edad (PA y MA, respectivamente), edad al 50% y 75% de madurez(I50% y I75%, respectivamente), peso adulto (β0), y otras características. Las heredabilidades directa ymaterna para PD fueron: 0.19 y 0.12, respectivamente. Las heredabilidades directas para PA, MA, I50%,I75% y β0 fueron 0.39, 0.15, 0.09, 0.20 y 0.09, respectivamente. La correlación genética para β0 y PA fue-0.47, indicando que al seleccionar animales por peso al año, tendríamos animales con mayor peso adulto.Las características de madurez pueden ser utilizadas en programas de mejoramiento genético en procura debúfalos mas eficientes en sistemas de producción colombianos.

Funções matemáticas não lineares propostas por Brody, Gompertz, Richards, Bertalanffy e Verhulstforam utilizadas para descrever o crescimento em búfalos provenientes de três regiões da Colômbia (Antioquia, Caldas e Córdoba). As funções que melhor descreveram o crescimento foram Brody e Gompertz.Com a base de dados da região de Caldas, foram estimadas as herdabilidades para as características pesoao desmame (PD), peso e maturidade ao ano de idade (PA e MA, respectivamente), idade ao 50% e 75% dematuridade (I50% e I75%, respectivamente), peso adulto (β0), e outras características. As herdabilidadesdireita e materna para PD foram: 0.19 e 0.12, respectivamente. As herdabilidades direitas para PA, MA,I50%, I75% e β0 foram 0.39, 0.15, 0.09, 0.20 e 0.09, respectivamente. A correlação genética para β0 e PA foi-0.47, indicando que ao selecionar animais por peso ao ano, teríamos animais com maior peso adulto. Ascaracterísticas de maturidade podem ser utilizadas em programas de melhoramento genético em procurade búfalos mais eficientes em sistemas de produção colombianos.

â-casein gene polymorphism permits identification of bovine milk mixed with bubaline milk in mozzarella cheese

Mozzarella cheese is traditionally prepared from bubaline (Bubalus bubalis) milk, but product adulteration occurs mainly by addition of or full substitution by bovine milk. The aim of this study was to show the usefulnes of molecular markers to identify the admixture of bovine milk to bubaline milk during the manufacturing process of mozzarella cheese. Samples of mozzarella cheese were produced by adding seven different concentrations of bovine milk: 0%, 1%, 2%, 5%, 8%, 12% and 100%. DNA extracted from somatic cells found in cheese were submitted to PCR-RFLP analysis of casein genes: alfa-s1-CN - CSN1S1 that encompasses 954 bp from exon VII to intron IX (Alu I and Hinf I), beta-CN - CSN2 including 495 bp of exon VII (Hae III and Hinf I), and k-CN - CSN3, encompassing 373 bp of exon IV (Alu I and Hind III). Our results indicate that Hae III-RFLP of CSN2 exon VII can be used as a molecular marker to detect the presence of bovine milk in "mozzarella" cheese.

Sequence characterization of coding regions of the myostatin gene ( GDF8 ) from Brazilian Murrah buffaloes ( Bubalus bubalis ) and comparison with the Bos taurus sequence

Within about 30 years the Brazilian buffalo (Bubalus bubalis) herd will reach approximately 50 million head as a result of the great adaptive capacity of these animals to tropical climates, together with the good productive and reproductive potential which make these animals an important animal protein source for poor and developing countries. The myostatin gene (GDF8) is important in the physiology of stock animals because its product produces a direct effect on muscle development and consequently also on meat production. The myostatin sequence is known in several mammalian species and shows a high degree of amino acid sequence conservation, although the presence of non-silent and silent changes in the coding sequences and several alterations in the introns and untranslated regions have been identified. The objective of our work was to characterize the myostatin coding regions of B. bubalis (Murrah breed) and to compare them with the Bos taurus regions looking for variations in nucleotide and protein sequences. In this way, we were able to identify 12 variations at DNA level and five alterations on the presumed myostatin protein sequence as compared to non double-muscled bovine sequences.

Responses to selection for milk traits in dairy buffaloes

The aim of the present study was to estimate the index and individual responses to selection for milk (MY), fat (FY) and protein (PY) yields for different breeding goals for two commercial buffalo milk production systems in São Paulo State characterized by: 1) all milk produced is sold to the industry (MILK) and 2) all milk produced is used in the mozzarella cheese-making process at the farm (MOZZARELLA). The current payment policy is based exclusively on milk volume. The mozzarella price refers to the wholesale selling price. Index responses to selection (IR) were calculated for three different breeding goals (BG): 1) MY exclusively (BG(1)); 2) FY + PY (BG(2)) and 3) MY + FY + PY (BG(3)). IR for the MILK system were 41.79 US dollars (BG(1)), 5.91 US dollars (BG(2)) and 38.22 US dollars (BG(3)). For the MOZZARELLA system, IR were 179.50 US dollars (BG(1)), 262.85 US dollars (BG(2)) and 402.41 US dollars (BG(3)). The results suggest that for the present circumstances, selection for milk components is not advantageous when milk is produced for sale to the industry. However, when mozzarella making is added to the system, the selection for components and milk volume is the most economically beneficial.

Kappa-casein gene study with molecular markers in female buffaloes (Bubalus bubalis)

Caseins comprise make up about 80 percent of the total protein content of milk and present polymorphism with changes in the amino acid sequence. Within this abundance of proteins, kappa-casein is noteworthy, since it has been associated with differences in milk yield, composition and processing. The objective of this study was to observe the existence of polymorphism in the kappa-casein gene in female buffaloes. For this purpose, blood samples from 115 female buffaloes, collected with vacutainer by needle punctionure of the jugular vein, were used. for genomic DNA extraction was done from blood samples. The PCR-RFLP and SSCP techniques demonstrated that the studied animals were monomorphic for the kappa-casein gene. Only allele B was observed in these animals, which was present in homozygosis. Therefore, it was not possible to quantify the gene action on milk yield and its constituents. The monomorphism observed in the population studied would allow the development of a method to identify mixtures of cow and buffalo milk in mozzarella cheese production, especially because, in cattle, the kappa-casein gene is polymorphic.

Biochemical and molecular investigations on qualitative and quantitative Hb polymorphism in the river buffalo (Bubalus bubalis L. ) population reared in Southern Italy

On 398 river buffalo samples, randomly collected in distinct breeding areas of the Campania region, high-resolution analytical systems were used to identify both qualitative and quantitative variations of the Hb phenotype. Polyacrylamide gel isoelectric focusing and HPLC were used to determine the ratio between HBA1 and HBA2 globin chains; restriction endonuclease analysis was performed to assess whether quantitative variations in Hb bands were related to an unusual number of a-globin genes. In the two buffalo subpopulations, allele frequencies of the alpha and beta globin systems were calculated, and F statistics (FIS, FIT and FST) were estimated as parameters of genetic diversity. The results suggest that: i) as shown by RFLP analysis, only a couple of associated a globin genes account for the quantitative variations recorded at the phenotypic level; ii) as expected, in the a globin gene system (HBA), the frequency of haplotype B (HBA-B) largely exceeded that of haplotype A (HBA-A) (95.1 percent vs 4.9 percent); iii) the frequency of the usual allele at the beta locus is 0.6, as opposed to 0.4 of the slow variant; iiii) the most significant component of variation of the genetic system of hemoglobin is between individuals within the same location.

An insight into the possible mechanism of working of two-cistronic gene expression systems and rational designing of newer systems.

The initial attempts at hyper-expressing buffalo/goat growth hormone (GH)-ORFs in Escherichia coli directly under various strong promoters were not successful despite the presence of a functional gene. High level expression of GH was achieved as a fusion protein with glutathione-S-transferase (GST). To produce native GH in an unfused state, we adapted an established strategy of two-cistronic approach in our system. In this strategy, utilizing one of the highly efficient reported sequences as the first cistron led to a nearly 1000-fold enhancement in the level of expression under an E. coli promoter (trc). In search of a newer first-cistron sequence as well as to see the generality of the two-cistronic approach, we explored the ability of different lengths of a highly expressing natural gene to act as an efficient first cistron. Surprisingly, GST, which is naturally highly expressible in E. coli, could not be fitted into a successful two-cistronic construct. In addition, placement of the entire two-cistronic expression cassette (which had earlier given high-level GH expression under trc promoter) under the T7 promoter in E. coli failed to hyper-express GH. These results suggest that the successful exploitation of the two-cistron arrangement for hyper-expression of eukaryotic ORFs in bacteria is not as straightforward as was previously thought. It appears probable that factors such as the sequence context, together with the length and codons used in the first cistron are important as well.

High salt method: a simple and rapid procedure for isolation of genomic DNA from buffalo (Bubalus bubalis) white blood cells.

Three different methods, namely guanidine hydrochloride, phenol-chloroform extraction and high salt method, were compared in order to develop a simple method for the extraction of high yield of DNA from buffalo blood suitable for genome analysis. Both phenol-chloroform and high salt methods produced good yields of high molecular weight DNA as determined by agarose gel electrophoresis. The yield and quality of DNA extracted by high salt method was comparable to that of phenol-chloroform method. The mean yields of DNA from 10 ml of whole blood extracted by either the phenol-chloroform or the high salt methods were 446.16 micrograms (SE = 26.68) and 432.83 micrograms (SE = 19.34) respectively. The DNA obtained from both methods was suitable for conventional as well as polymerase chain reaction (PCR) based restriction fragment length polymorphism (RFLP) studies. Extraction using the guanidine hydrochloride method resulted in a gelatinous material that failed to resuspend in TE buffer. The high salt method is quick and reliable and can be routinely used for the extraction of DNA from buffalo samples instead of phenol-chloroform extraction which is hazardous and time-consuming.