Prostaglandin F2α treatment concurrent with artificial insemination does not affect bovine embryo production / Administração de prostaglandina F2 alfa no momento da inseminação artificial não afeta a produção de embriões bovinos

ABSTRACT: Treatment with prostaglandin F2α (PGF) induces ovulation and increases conception rates in cows, while improving embryo production in buffalos. However, its effect on superovulated cows is unknown. This study verified whether single PGF administration concurrent with artificial insemination (AI) improves fertilization and embryo production rates in superovulated cows. In each replicate, embryo donor cows were equally allocated to two groups: the untreated control and PGF groups. The latter of which received 482 µg of cloprostenol concurrent with the first AI. Each cow (n = 35) was subjected to two superovulations (SOV) in a crossover design (total = 70 embryo collections). In the control and PGF groups, respectively, the observed responses were [median (95% CI)]: 12 (10-18) and 15 (12-18) total structures, 9 (7-11) and 7 (6-10) viable embryos, 1 (0-1) and 1 (1-3) degenerated embryos, and 1 (0-3) and 2 (0-5) oocytes (P > 0.05). In conclusion, single PGF treatment concurrent with the first AI did not affect embryo production in superovulated cows.

RESUMO: A prostaglandina F2α (PGF) pode induzir a ovulação e melhorar tanto a concepção em vacas, como a produção de embriões em búfalas, mas o efeito em vacas superovuladas é desconhecido. Esse estudo teve como objetivo verificar se a administração de uma dose de PGF na inseminação artificial (IA) após a superovulação (SOV) melhora as taxas de fecundação e produção embrionária em vacas. Em cada replicação, vacas doadoras de embriões foram equilibradamente alocadas em dois grupos: controle, não tratado, ou PGF, que recebeu 482 µg de cloprostenol no momento da primeira IA. Cada doadora (n = 35) foi submetida a duas SOV em um delineamento crossover (total = 70 coletas de embriões). Nos grupos controle e PGF, respectivamente, foram observados [medianas (IC 95%)]: 12 (10-18) e 15 (12-18) estruturas totais; 9 (7-11) e 7 (6-10) embriões viáveis; 1 (0-1) e 1 (1-3) embriões degenerados; e 1 (0-3) e 2 (0-5) ovócitos (P > 0,05). Conclui-se que uma única administração de PGF no momento da primeira IA não afeta a produção embrionária de vacas superovuladas.

Prostaglandin F2α treatment concurrent with artificial insemination does not affect bovine embryo production / Administração de prostaglandina F2 alfa no momento da inseminação artificial não afeta a produção de embriões bovinos

Treatment with prostaglandin F2α (PGF) induces ovulation and increases conception rates in cows, while improving embryo production in buffalos. However, its effect on superovulated cows is unknown. This study verified whether single PGF administration concurrent with artificial insemination (AI) improves fertilization and embryo production rates in superovulated cows. In each replicate, embryo donor cows were equally allocated to two groups: the untreated control and PGF groups. The latter of which received 482 µg of cloprostenol concurrent with the first AI. Each cow (n = 35) was subjected to two superovulations (SOV) in a crossover design (total = 70 embryo collections). In the control and PGF groups, respectively, the observed responses were [median (95% CI)]: 12 (10-18) and 15 (12-18) total structures, 9 (7-11) and 7 (6-10) viable embryos, 1 (0-1) and 1 (1-3) degenerated embryos, and 1 (0-3) and 2 (0-5) oocytes (P > 0.05). In conclusion, single PGF treatment concurrent with the first AI did not affect embryo production in superovulated cows.

A prostaglandina F2α (PGF) pode induzir a ovulação e melhorar tanto a concepção em vacas, como a produção de embriões em búfalas, mas o efeito em vacas superovuladas é desconhecido. Esse estudo teve como objetivo verificar se a administração de uma dose de PGF na inseminação artificial (IA) após a superovulação (SOV) melhora as taxas de fecundação e produção embrionária em vacas. Em cada replicação, vacas doadoras de embriões foram equilibradamente alocadas em dois grupos: controle, não tratado, ou PGF, que recebeu 482 µg de cloprostenol no momento da primeira IA. Cada doadora (n = 35) foi submetida a duas SOV em um delineamento crossover (total = 70 coletas de embriões). Nos grupos controle e PGF, respectivamente, foram observados [medianas (IC 95%)]: 12 (10-18) e 15 (12-18) estruturas totais; 9 (7-11) e 7 (6-10) embriões viáveis; 1 (0-1) e 1 (1-3) embriões degenerados; e 1 (0-3) e 2 (0-5) ovócitos (P > 0,05). Conclui-se que uma única administração de PGF no momento da primeira IA não afeta a produção embrionária de vacas superovuladas.

Accidental monensin poisoning in buffaloes in Bahia, Brazil

ABSTRACT: Monensin is an ionophore antibiotic (IA) widely used for growth promotion and weight gain in the production of ruminants. However, it has caused intoxication in several species, including buffaloes, mainly because of the ignorance or disrespect of the recommendations for use in each animal species. The objective of this study was to describe, for the first time, clinical-epidemiological and anatomopathological data of an outbreak of accidental poisoning by monensin in buffalos and rediscuss the recommendation of the use of IA in the production of this species. The outbreak affected 21 adult buffaloes after consumption of remains from a feed formulated on the farm and whose constituents were mixed by hand. Clinical and first death signs were observed 24 hours after ingestion of this food. In general, the clinical picture was characterized by muscle weakness, tremors, difficulty in locomotion, and decubitus. Fifteen buffaloes presented clinical signs of poisoning (71.5% morbidity), followed by death (100% lethality), after acute to subacute evolution ( 24h to 96h). Laboratory tests indicated elevated serum activity of creatine phosphokinase and aspartate aminotransferase enzymes. Three buffaloes underwent necropsy, and samples from several organs were collected for histopathological examination. The main injuries found were hyaline degeneration and multifocal segmental necrosis in the skeletal and cardiac striated muscles (myopathy and degenerative-necrotic multifocal multifocal-necrotic cardiopathy). The diagnosis was confirmed by the toxicological evaluation of suspected ration remains, which detected 461.67mg/kg of monensin. The death of 71.5% buffaloes in this lot occurred due to a succession of errors, which included faults in the formulation of the ration and, above all, due to the use of monensin in a highly sensitive species. Despite the possible beneficial effects of IA use as a dietary supplement for buffaloes, we are of the opinion that IAs should never be used in bubalinoculture since any increment in production does not compensate for the imminent risk of death due to a small safety margin for this species and the absence of antidotes.

RESUMO: Monensina é um antibiótico ionóforo (AI) amplamente empregado na produção de ruminantes para promoção de crescimento e ganho de peso, mas que tem causado intoxicação em diversas espécies, incluindo os búfalos, principalmente, pelo desconhecimento ou desrespeito das recomendações de uso e às particularidades de cada espécie animal. Objetivou-se descrever, pela primeira vez na Bahia, dados clínico-epidemiológicos e anatomopatológicos de um surto de intoxicação acidental por monensina em búfalos e rediscutir a recomendação do uso de AI na produção de bubalinos. O surto acometeu um lote de 21 búfalos adultos após consumo de sobras de uma ração para bovinos formulada na fazenda e cujos constituintes eram misturados à mão. Os sinais clínicos e primeiros óbitos foram observados 24 horas após a ingestão dessa ração. O quadro clínico, em geral, se caracterizou por fraqueza muscular, tremores, dificuldade de locomoção e decúbito. Quinze búfalos apresentaram sinais clínicos de intoxicação (morbidade 71,5%), seguido de morte (letalidade 100%), após evolução aguda a subaguda ( 24h até 96h). Exames laboratoriais indicaram acentuada elevação na atividade sérica das enzimas CPK e AST. Três búfalos foram necropsiados, sendo coletadas amostras de diversos órgãos para exame histopatológico. A principal lesão encontrada foi degeneração hialina e necrose segmentar multifocal nos músculos estriados esqueléticos e cardíacos (miopatia e cardiopatia degenerativo-necrótica tóxica multifocal polifásica). O diagnóstico foi confirmado pela avaliação toxicológica das sobras da ração suspeita, que detectou 461,67mg/kg de monensina. A morte de 71,5% dos búfalos deste lote ocorreu devido a uma sucessão de erros, que incluíram falhas na formulação da ração e, sobretudo, devido ao uso da monensina em uma espécie altamente sensível. Enfatizamos que, apesar dos possíveis efeitos benéficos do uso AIs como suplemento dietético para bubalinos, somos da opinião que os AIs nunca devem ser empregados na bubalinocultura, uma vez que os eventuais incrementos na produção não compensam o risco iminente de morte, devido a pequena margem de segurança para essa espécie e a inexistência de antídotos.

Innate immunity gene expression profiles in conjunctival membrane biopsies from Amazonian buffalo / Perfis de expressão de genes da imunidade inata em biopsias de membrana conjuntival de búfalos da Amazônia

Domestic buffalo production plays an economically important role in the Brazilian Amazon, but they are susceptible to many diseases favored by the tropical climate and annually flooded habitats, including ocular diseases. In this context, it is important to select genotypes that maximize innate ocular immunity in Amazonian herds. We aimed to characterise, for the first time, gene expression profiles of the innate immune system in the conjunctival membrane of buffalo. Ocular conjunctival tissue samples were collected from 60 clinically healthy slaughtered animals in the northern Brazilian state of Amapá. The samples were histologically processed for classification into three groups according to the quantitative degree of lymphoid tissue associated with the conjunctiva (discrete, G1; slight, G2; and moderate, G3 presence of lymphoid tissue). RT-PCR was used to quantify gene expression of inflammatory cytokine (IL6, IL10, TNFA, IFNG), Toll-like receptor 4 (TLR4), and Defensin beta 110 (DEFB110), relative to the endogenous GAPDH gene. G1 animals presented low expression for IL6, IL10, TNFA, and DEFB110, while G2 exhibited high expression for IL6, IL10, IFNG, and TLR4. All G3 animals showed high expression for all tested genes. These results suggest a greater resistance to pathogenic microorganisms of buffalos in the G3 group, and the proportion of lymphoid tissue associated with the conjunctiva may be related to the immune resistance of individuals.(AU)

A produção de búfalos domésticos desempenha um papel economicamente importante na Amazônia brasileira, mas eles são suscetíveis a muitas doenças favorecidas pelo clima tropical e habitats inundados anualmente, incluindo doenças oculares. Nesse contexto, é importante selecionar genótipos que maximizem a imunidade ocular inata em rebanhos amazônicos. Objetivamos caracterizar, pela primeira vez, perfis de expressão gênica do sistema imune inato na membrana conjuntival de búfalos. Amostras de tecido conjuntival ocular foram coletadas de 60 animais clinicamente saudáveis abatidos no estado do Amapá, norte do Brasil. As amostras foram processadas histologicamente para classificação em três grupos de acordo com o grau quantitativo de tecido linfoide associado à conjuntiva (discreta, G1; leve, G2; e moderada, G3 presença de tecido linfoide). RT-PCR foi utilizado para quantificar a expressão gênica de citocinas inflamatórias (IL6, IL10, TNFA, IFNG), receptor Toll-like 4 (TLR4) e Defensina beta 110 (DEFB110), em relação ao gene GAPDH endógeno. Os animais do G1 apresentaram baixa expressão para IL6, IL10, TNFA e DEFB110, enquanto G2 exibiu alta expressão para IL6, IL10, IFNG e TLR4. Todos os animais do G3 apresentaram alta expressão para todos os genes testados. Esses resultados sugerem maior resistência aos microrganismos patogênicos dos búfalos do grupo G3, e a proporção de tecido linfoide associado à conjuntiva pode estar relacionada à resistência imunológica dos indivíduos.(AU)

Leptospirosis, bovine viral diarrhea and infectious bovine rhinotracheitis: prevalence in Colombian cattle and buffaloes

One of the limiting factors of productive efficiency in cattle and buffalo herds is related to the high prevalence of infectious diseases which affect reproduction. The aim of this study was to determine the prevalence of anti-Leptospiraantibodies for bovine viral diarrhea virus (BVDV) and bovine herpesvirus type 1 (BoHV-1) in bovine and buffalo herds in Colombia. Blood serum samples were collected from 1100 buffaloes and 1000 cattle. The ELISA technique was used to detect antibodies against BVDV and BoHV-1, and the microscopic agglutination technique to detect anti-Leptospiraantibodies. The prevalence of anti-Leptospiraantibodies and of BVDV and BoHV-1 in bovine samples was observed in 16, 39.7, and 65% of animals, respectively, while the positivity in samples for the same antibodies in buffalos was detected in 18.7, 27.5 and 51.5%, respectively. Exposure of cattle and buffaloes to BoHV-1 was positively associated with age, higher prevalence rates were observed in older ages. Seropositivity of cattle for BVDV and BoHV-1 was higher in male animals. Activities such as embryo transfer, milking, and needle reuses, as well as the presence of cats and rodents are factors which favor positivity of the herd for BVDV and BoHV-1.(AU)

Accidental monensin poisoning in buffaloes in Bahia, Brazil / Intoxicação acidental por monensina em búfalos na Bahia, Brasil

Monensin is an ionophore antibiotic (IA) widely used for growth promotion and weight gain in the production of ruminants. However, it has caused intoxication in several species, including buffaloes, mainly because of the ignorance or disrespect of the recommendations for use in each animal species. The objective of this study was to describe, for the first time, clinical-epidemiological and anatomopathological data of an outbreak of accidental poisoning by monensin in buffalos and rediscuss the recommendation of the use of IA in the production of this species. The outbreak affected 21 adult buffaloes after consumption of remains from a feed formulated on the farm and whose constituents were mixed by hand. Clinical and first death signs were observed 24 hours after ingestion of this food. In general, the clinical picture was characterized by muscle weakness, tremors, difficulty in locomotion, and decubitus. Fifteen buffaloes presented clinical signs of poisoning (71.5% morbidity), followed by death (100% lethality), after acute to subacute evolution (<24h to 96h). Laboratory tests indicated elevated serum activity of creatine phosphokinase and aspartate aminotransferase enzymes. Three buffaloes underwent necropsy, and samples from several organs were collected for histopathological examination. The main injuries found were hyaline degeneration and multifocal segmental necrosis in the skeletal and cardiac striated muscles (myopathy and degenerative-necrotic multifocal multifocal-necrotic cardiopathy). The diagnosis was confirmed by the toxicological evaluation of suspected ration remains, which detected 461.67mg/kg of monensin. The death of 71.5% buffaloes in this lot occurred due to a succession of errors, which included faults in the formulation of the ration and, above all, due to the use of monensin in a highly sensitive species. Despite the possible beneficial effects of IA use as a dietary supplement for buffaloes, we are of the opinion that IAs should never be used in bubalinoculture since any increment in production does not compensate for the imminent risk of death due to a small safety margin for this species and the absence of antidotes.

Monensina é um antibiótico ionóforo (AI) amplamente empregado na produção de ruminantes para promoção de crescimento e ganho de peso, mas que tem causado intoxicação em diversas espécies, incluindo os búfalos, principalmente, pelo desconhecimento ou desrespeito das recomendações de uso e às particularidades de cada espécie animal. Objetivou-se descrever, pela primeira vez na Bahia, dados clínico-epidemiológicos e anatomopatológicos de um surto de intoxicação acidental por monensina em búfalos e rediscutir a recomendação do uso de AI na produção de bubalinos. O surto acometeu um lote de 21 búfalos adultos após consumo de sobras de uma ração para bovinos formulada na fazenda e cujos constituintes eram misturados à mão. Os sinais clínicos e primeiros óbitos foram observados 24 horas após a ingestão dessa ração. O quadro clínico, em geral, se caracterizou por fraqueza muscular, tremores, dificuldade de locomoção e decúbito. Quinze búfalos apresentaram sinais clínicos de intoxicação (morbidade 71,5%), seguido de morte (letalidade 100%), após evolução aguda a subaguda (<24h até 96h). Exames laboratoriais indicaram acentuada elevação na atividade sérica das enzimas CPK e AST. Três búfalos foram necropsiados, sendo coletadas amostras de diversos órgãos para exame histopatológico. A principal lesão encontrada foi degeneração hialina e necrose segmentar multifocal nos músculos estriados esqueléticos e cardíacos (miopatia e cardiopatia degenerativo-necrótica tóxica multifocal polifásica). O diagnóstico foi confirmado pela avaliação toxicológica das sobras da ração suspeita, que detectou 461,67mg/kg de monensina. A morte de 71,5% dos búfalos deste lote ocorreu devido a uma sucessão de erros, que incluíram falhas na formulação da ração e, sobretudo, devido ao uso da monensina em uma espécie altamente sensível. Enfatizamos que, apesar dos possíveis efeitos benéficos do uso AIs como suplemento dietético para bubalinos, somos da opinião que os AIs nunca devem ser empregados na bubalinocultura, uma vez que os eventuais incrementos na produção não compensam o risco iminente de morte, devido a pequena margem de segurança para essa espécie e a inexistência de antídotos.

Accidental monensin poisoning in buffaloes in Bahia, Brazil / Intoxicação acidental por monensina em búfalos na Bahia, Brasil

Monensin is an ionophore antibiotic (IA) widely used for growth promotion and weight gain in the production of ruminants. However, it has caused intoxication in several species, including buffaloes, mainly because of the ignorance or disrespect of the recommendations for use in each animal species. The objective of this study was to describe, for the first time, clinical-epidemiological and anatomopathological data of an outbreak of accidental poisoning by monensin in buffalos and rediscuss the recommendation of the use of IA in the production of this species. The outbreak affected 21 adult buffaloes after consumption of remains from a feed formulated on the farm and whose constituents were mixed by hand. Clinical and first death signs were observed 24 hours after ingestion of this food. In general, the clinical picture was characterized by muscle weakness, tremors, difficulty in locomotion, and decubitus. Fifteen buffaloes presented clinical signs of poisoning (71.5% morbidity), followed by death (100% lethality), after acute to subacute evolution (<24h to 96h). Laboratory tests indicated elevated serum activity of creatine phosphokinase and aspartate aminotransferase enzymes. Three buffaloes underwent necropsy, and samples from several organs were collected for histopathological examination. The main injuries found were hyaline degeneration and multifocal segmental necrosis in the skeletal and cardiac striated muscles (myopathy and degenerative-necrotic multifocal multifocal-necrotic cardiopathy). The diagnosis was confirmed by the toxicological evaluation of suspected ration remains, which detected 461.67mg/kg of monensin. The death of 71.5% buffaloes in this lot occurred due to a succession of errors, which included faults in the formulation of the ration and, above all, due to the use of monensin in a highly sensitive species. Despite the possible beneficial effects of IA use as a dietary supplement for buffaloes, we are of the opinion that IAs should never be used in bubalinoculture since any increment in production does not compensate for the imminent risk of death due to a small safety margin for this species and the absence of antidotes.(AU)

Monensina é um antibiótico ionóforo (AI) amplamente empregado na produção de ruminantes para promoção de crescimento e ganho de peso, mas que tem causado intoxicação em diversas espécies, incluindo os búfalos, principalmente, pelo desconhecimento ou desrespeito das recomendações de uso e às particularidades de cada espécie animal. Objetivou-se descrever, pela primeira vez na Bahia, dados clínico-epidemiológicos e anatomopatológicos de um surto de intoxicação acidental por monensina em búfalos e rediscutir a recomendação do uso de AI na produção de bubalinos. O surto acometeu um lote de 21 búfalos adultos após consumo de sobras de uma ração para bovinos formulada na fazenda e cujos constituintes eram misturados à mão. Os sinais clínicos e primeiros óbitos foram observados 24 horas após a ingestão dessa ração. O quadro clínico, em geral, se caracterizou por fraqueza muscular, tremores, dificuldade de locomoção e decúbito. Quinze búfalos apresentaram sinais clínicos de intoxicação (morbidade 71,5%), seguido de morte (letalidade 100%), após evolução aguda a subaguda (<24h até 96h). Exames laboratoriais indicaram acentuada elevação na atividade sérica das enzimas CPK e AST. Três búfalos foram necropsiados, sendo coletadas amostras de diversos órgãos para exame histopatológico. A principal lesão encontrada foi degeneração hialina e necrose segmentar multifocal nos músculos estriados esqueléticos e cardíacos (miopatia e cardiopatia degenerativo-necrótica tóxica multifocal polifásica). O diagnóstico foi confirmado pela avaliação toxicológica das sobras da ração suspeita, que detectou 461,67mg/kg de monensina. A morte de 71,5% dos búfalos deste lote ocorreu devido a uma sucessão de erros, que incluíram falhas na formulação da ração e, sobretudo, devido ao uso da monensina em uma espécie altamente sensível. Enfatizamos que, apesar dos possíveis efeitos benéficos do uso AIs como suplemento dietético para bubalinos, somos da opinião que os AIs nunca devem ser empregados na bubalinocultura, uma vez que os eventuais incrementos na produção não compensam o risco iminente de morte, devido a pequena margem de segurança para essa espécie e a inexistência de antídotos.(AU)

Evaluation of automated monitoring calving prediction in dairy buffaloes a new tool for calving management / Avaliação do monitoramento automatizado da previsão de parto em búfalos leiteiros: uma nova ferramenta para o gerenciamento de parto

Buffalo is one of the leading milk-producing dairy animals. Its production and reproduction are affected due to some factors including inadequate monitoring around parturition, which cause economic losses like delayed birth process, increased risk of stillbirth, etc. The appropriate calving monitoring is essential for dairy herd management. Therefore, we designed a study its aim was, to predict the calving based on automated machine measured prepartum behaviors in buffaloes. The data were collected from n=40 pregnant buffaloes of 2nd to 5th parity, which was synchronized. The NEDAP neck and leg logger tag was attached to each buffalo at 30 days before calving and automatically collected feeding, rumination, lying, standing, no. of steps, no. of switches from standing to lying (lying bouts) and total motion activity. All behavioral data were reduced to -10 days before the calving date for statistical analysis to use mixed model procedure and ANOVA. Results showed that feeding and rumination time significantly (P<0.05) decreased from -10 to -1 days before calving indicating calving prediction. Moreover, Rumination time was at lowest (P<0.001) value at 2h before the calving such behavioral changes may be useful to predict calving in buffaloes. Similarly, lying bouts and standing time abruptly decreased (P<0.05) from -3 to -1 days before calving, while lying time abruptly increased (P<0.01) from -3 to -1 days before calving (531.57±23.65 to 665.62±18.14, respectively). No. of steps taken and total motion significantly (P<0.05) increased from -10 to -1 days before calving. Feeding time was significantly (P<0.02) lowered in 3rd parity buffaloes compared with 2nd, 4th and 5th parity buffaloes, while standing time of 5th parity buffaloes were lowered (P<0.05) as compared to 2nd to 4th parity buffalos at -1 day of prepartum. However, rumination, lying, no. of steps taken and total motion activity at -1 day of prepartum was independent (P>0.05) of parity in buffaloes. Neural network analysis for combined variables from NEDAP technology at the daily level yielded 100.0% sensitivity and 98% specificity. In conclusion NEDAP technology can be used to measured behavioral changes -10 day before calving as it can serve as a useful guide in the prediction calving date in the buffaloes.(AU)

O búfalo é um dos principais animais produtores de leite. Sua produção e sua reprodução são afetadas por causa de alguns fatores, incluindo o monitoramento inadequado ao redor do parto, que causam perdas econômicas, como atraso no processo de parto, aumento do risco de natimorto, etc. O monitoramento adequado do parto é essencial para o manejo do rebanho leiteiro. Portanto, projetamos um estudo cujo objetivo foi prever o parto com base em comportamentos pré-parto medidos por máquina automatizada em búfalas. Os dados foram coletados de 40 búfalas prenhes de 2ª a 5ª paridade, que foi sincronizada. A etiqueta NEDAP de pescoço e perna foi fixada em cada búfala 30 dias antes do parto e coletava dados, automaticamente, durante a alimentação e a ruminação, em posição deitada e em pé, além do número de passos, número de mudanças de pé para deitado (período deitado) e atividade de movimento total. Todos os dados comportamentais foram reduzidos para -10 dias antes da data do parto para análise estatística usando o procedimento de modelo misto e ANOVA. Os resultados mostraram que o tempo de alimentação e de ruminação diminuiu significativamente (P<0,05) de -10 dias para -1 dia antes do parto, indicando a previsão de parto. Além disso, o tempo de ruminação apresentou seu menor valor (P<0,001) 2 horas antes do parto, e tais mudanças comportamentais podem ser úteis para predizer o parto em búfalas. Da mesma forma, o período deitado e o tempo em pé diminuíram abruptamente (P<0,05) de -3 dias para -1 dia antes do parto, enquanto o tempo deitado aumentou abruptamente (P<0,01) de -3 dias para -1 dia antes do parto (531,57 ± 23,65 para 665,62± 18,14, respectivamente). O número de passos dados e o movimento total aumentaram significativamente (P<0,05) de -10 para -1 dias antes do parto. O tempo de alimentação foi significativamente (P<0,02) reduzido em búfalas de 3ª paridade em comparação com búfalas de 2ª, 4ª e 5ª paridade, enquanto o tempo de espera de búfalas de 5ª paridade foi reduzido (P<0,05) em comparação com búfalas de 2ª a 4ª paridade em -1 dia antes do parto. No entanto, ruminação, posição deitada, número de passos dados e atividade de movimento total em -1 dia antes do parto foram independentes (P>0,05) da paridade em búfalas. A análise de rede neural para variáveis ​​combinadas da tecnologia NEDAP no nível diário produziu 100% de sensibilidade e 98% de especificidade. Em conclusão, a tecnologia NEDAP pode ser usada para medir mudanças comportamentais -10 dias antes do parto, pois pode servir como um guia útil para prever a data do parto em búfalas.(AU)

Intoxicação por lasalocida em bezerros búfalos no Estado do Pará, Brasil / Lasalocid poisoning in buffalo calves in the state of Pará, Brazil

Background: Ionophore antibiotics are food additives with coccidiostatic or antimicrobial action; they are also used as growth promoters, ruminal pH regulators, volatile fatty acid molar modifiers, and methanogenesis reducers. However, these compounds have the potential to cause microbial resistance, in addition to the risk of intoxication. Ionophore poisoning may be caused by excessive intake, sensitivity of certain animal species, and concomitant use with other drugs. In Brazil, cases of ionophore poisoning in buffalos are rare. This study aims to describe the epidemiological, clinical, and pathological findings of lasalocid poisoning in buffalo calves. Case: A visit was made to a farm in the municipality of Mojú, Pará state to care for Murrah buffalo calves. After weaning, the buffalos were grazed in paddocks with Panicum spp., and received a supplement of mineral, protein, and vitamin. This supplement contained, per kg, 250 g PB, 50 g Ca, 20 g P, 8 g S, 39 g Na, 20 mg Co, 557 mg Cu, 200 mg Fe, 12.4 mg Se, 2040 mg Zn, 0.19 mg biotin, 26750 IU of vitamin A, 4175 IU of vitamin D, 155 IU of vitamin E and 300 mg/kg of lasalocid. The product was made available to all calves, at 1-2 g/kg body weight (BW), according to the manufacturer's recommendations. Older calves were raised together with those less than 30 days old; as a result, the older calves tended to eat more, which could lead to a supplementation consumption of more than 1 kg body weight per animal per day. It was reported that between 40 and 60 days after the introduction of this supplement, 16 calves fell ill and died due to apathy, motor instability, tremors, and distended neck. The herd had a mortality rate of 33.3%. Two calves underwent a necroscopic examination at the Pathology Section of the Veterinary Institute of the Federal University of Pará. Macroscopic examination revealed extensive pale areas in the skeletal muscles, myocardium, and tongue. Fragments of these muscles and various organs were collected, fixed in 10% buffered formalin, processed according to the routine histological technique, and stained with hematoxylin-eosin and Masson's trichrome stain. Microscopic examination of the histologic samples revealed foci of muscle atrophy and necrosis characterized by an increase in cytoplasmic eosinophilia associated with the loss of stretch marks, and hyperchromatic nuclei that were displaced to the periphery. The necrosis of the muscle fibers was highlighted by Masson's trichrome staining. Discussion: The diagnosis of lasalocid poisoning in buffalo calves was based on epidemiological data, clinical findings, results of macroscopic and histopathological examination, and based on the estimated ionophore intake, obtained directly from the supplement label and by the calf's handler. Based on the absence of stratification of the calves by similarity of age and because the buffalo calves older than 30 days could eat more than 1 kg of the supplement (containing 300 mg/kg of lasalocid), it was possible to estimate the intake of lasalocid per kg CP (body weight). Therefore, the intake of lasalocid by a 70-kg buffalo calf in approximately 90 days and daily supplement consumption between 1 and 1.5 kg would be between 4.2 and 6.4 mg/kg of body weight. This report reinforces that notion that buffalo calves should never ingest ionophores; however, if necessary, strict protocols must be followed to avoid poisoning in these animals. This study highlighted the fact that stratification of buffaloes by different age groups during feeding became a risk factor that allowed greater consumption by older animals; this led to the estimated consumption of 4.2-6.4 mg/kg of lasalocid.

Viabilidade do uso de dietas com grão total para bubalinos / Feasibility of using full grain diets for buffalos

O objetivo do trabalho foi avaliar o consumo de matéria seca (MS), desempenho, e viabilidade econômica de búfalos submetidos a dietas de grão total comparado como de dietas convencionais. Doze bubalinos foram alimentados em confinamento e os tratamentos dietéticos foram oferecidos por 63 dias de período de comparação. Os tratamentos consistiram em dieta com 100% de concentrado (Grão Total) e dieta com 70% de concentrado mais 30% de silagem de milho (Convencional) com base MS. Amostras dos ingredientes e das sobras alimentares de cada animal foram coletadas, para determinação da matéria seca, e pesadas, para cálculo do consumo individual e da eficiência alimentar. Foi realizada coleta de sangue, por dezoito horas, para determinação de NUP (Nitrogênio Uréico plasmático). O peso vivo (PV) dos bubalinos foi determinado no início do período experimental e a cada sete dias de comparação. O ganho diário de peso foi calculado para cada uma das nove semanas e a viabilidade econômica das duas dietas, através do custo alimentar, avaliada em uma planilha do Excel®. Animais alimentados com dieta grão total apresentaram menor consumo de matéria seca (CMS kg e CMS % PV) que animais alimentados com dieta convencional. O peso vivo final (PVF), o peso da carcaça, e o ganho de peso médio diário (GMD) dos animais em kg, não diferiram estatisticamente entre os tratamentos. Em relação ao rendimento de carcaça, animais alimentados com grão total diferiram estatisticamente. O NUP, após a primeira alimentação, foi maior em animais alimentados com dieta convencional. Os bubalinos alimentados com grão total apresentaram menor custo alimentar e maior rendimento de carcaça.

The objective of this work was to evaluate dry matter consumption, performance, and economic viability of buffaloes submitted to full grain diets compared to conventional diets. Twelve non-castrated buffalo of the Murah breed were fed in confinement. Dietary treatments were offered for a 63-day comparison period. The treatments consisted of a diet with 100% concentrate (grain exclusive diet) and a diet with 70% concentrate plus 30% corn silage (Conventional) based on dry matter. Samples of the ingredients and food leftovers from each animal were collected to determine dry and heavy matter, to calculate individual consumption and feed efficiency. Blood was collected for 18 hours to determine PUN (plasma urea nitrogen). The live weight (LW) of the buffaloes was determined at the beginning of the experimental period and every seven days for comparison. The daily weight gain was calculated for each of the nine weeks and the economic viability of the two diets, through the food cost was evaluated in Excel® spreadsheet. Animals fed with a grain exclusive diet had lower consumption of dry matter (DMI lb and DMI % LW) than animals fed with conventional diet. The final live weight (LLW), carcass weight, average daily weight gain (PDG) of the animals in lb, did not differ statistically between treatments. Regarding the carcass yield, animals fed with total grain differed statistically. The PUN after the first feeding was higher in animals fed with a conventional diet. The buffaloes fed with total grain showed lower food cost and higher carcass yield.

Estudo soroepidemiológico de alphaherpesvírus bovino 1 (BoHV-1) em bubalinos (Bubalus bubalis) de corte da Região Sul do Brasil / Seroepidemiology survey of bovine alphaherpesvirus 1 (BoHV-1) in unvaccinated beef bubalines (Bubalus bubalis) from Southern Brazil

Background: The bovine alphaherpesvirus type 1 (BoHV-1) belonging to the order Herpesvirales, family Herpesviridae,subfamily Alphaherpesvirinae and genus Varicellovirus. Bovine alphaherpesvirus 1 (BoHV-1) infections have a great importance due to the high rates of dissemination in cattles worldwide. Although, the BoHV-1 was largely related in cattle,buffaloes were also classified as host for the virus. However, studies that determine seroepidemiological data in this species are scarce and necessary. The aim of the present study was to determine the presence of antibodies against BoHV-1in healthy beef buffaloes using the virus neutralization (VN) technique.Cases: This work portrays an epidemiological survey, based on a sectional study characterized by blood samples collectedfrom 54 Murrah buffalo, aged 6 to 24 months, from the municipalities of Guaraqueçaba, Ponta Grossa, Antonina andDoutor Ulysses, located in Paraná state, being 20, 14, 10 and 10 samples from each location, respectively. Thirty-sevenpercent (20/54) of the samples were collected at slaughterhouse with registration at the Federal Inspection Service (SIF)of the Ministry of Agriculture, Livestock and Food Supply (MAPA), and 63% (34/54) at the buffalo’s farms. The serumsamples were collected in sterile tubes without anticoagulant in stored isothermal boxes, with serum separation and storedat -20ºC. The samples were sent to the Animal Virology Laboratory of Universidade Estadual de Londrina, UEL for serological analysis. Serological diagnosis using the virus neutralization (VN) technique was performed according the OIEmanual of diagnostic tests and vaccines for terrestrial animals, using the BoHV-1 Los Angeles strain. Forth-five of the 54samples (83.3%) evaluated generated titers of antibodies against BoHV-1 present in all evaluated herds, and the samplescollected in herd from Antonina city were 100% positive, following by 80% in Guaraqueçaba...

Estudo soroepidemiológico de alphaherpesvírus bovino 1 (BoHV-1) em bubalinos (Bubalus bubalis) de corte da Região Sul do Brasil / Seroepidemiology survey of bovine alphaherpesvirus 1 (BoHV-1) in unvaccinated beef bubalines (Bubalus bubalis) from Southern Brazil

Background: The bovine alphaherpesvirus type 1 (BoHV-1) belonging to the order Herpesvirales, family Herpesviridae,subfamily Alphaherpesvirinae and genus Varicellovirus. Bovine alphaherpesvirus 1 (BoHV-1) infections have a great importance due to the high rates of dissemination in cattles worldwide. Although, the BoHV-1 was largely related in cattle,buffaloes were also classified as host for the virus. However, studies that determine seroepidemiological data in this species are scarce and necessary. The aim of the present study was to determine the presence of antibodies against BoHV-1in healthy beef buffaloes using the virus neutralization (VN) technique.Cases: This work portrays an epidemiological survey, based on a sectional study characterized by blood samples collectedfrom 54 Murrah buffalo, aged 6 to 24 months, from the municipalities of Guaraqueçaba, Ponta Grossa, Antonina andDoutor Ulysses, located in Paraná state, being 20, 14, 10 and 10 samples from each location, respectively. Thirty-sevenpercent (20/54) of the samples were collected at slaughterhouse with registration at the Federal Inspection Service (SIF)of the Ministry of Agriculture, Livestock and Food Supply (MAPA), and 63% (34/54) at the buffalos farms. The serumsamples were collected in sterile tubes without anticoagulant in stored isothermal boxes, with serum separation and storedat -20ºC. The samples were sent to the Animal Virology Laboratory of Universidade Estadual de Londrina, UEL for serological analysis. Serological diagnosis using the virus neutralization (VN) technique was performed according the OIEmanual of diagnostic tests and vaccines for terrestrial animals, using the BoHV-1 Los Angeles strain. Forth-five of the 54samples (83.3%) evaluated generated titers of antibodies against BoHV-1 present in all evaluated herds, and the samplescollected in herd from Antonina city were 100% positive, following by 80% in Guaraqueçaba...(AU)

Use of estradiol cypionate to induce ovulation in buffalo raised in floodplain areas of Itacoatiara, Amazonas / [Uso do cipionato de estradiol na indução da ovulação de búfalas criadas em várzea da região de Itacoatiara, Amazonas]

This study aimed to determine the efficiency of estradiol cypionate (EC) as an ovulation inducer in a Timed Artificial Insemination protocol. 69 buffalo cows received an intravaginal progesterone device and 2mg of estradiol benzoate (EB) at D0. On D9, the intravaginal device was removed and 0.53mg of prostaglandin (PGF2α) and 400UI of equine chorionic gonadotrophin (eCG) were applied. The cows were distributed into two groups: the first group received 1mg of EC (ECG) in D9, and the second group received 1mg of EB (EBG) in D10. Inseminations occurred on D11. Ovarian activity and pregnancy diagnosis were analyzed by ultrasonography. There was no difference (P>0.05) in follicular diameter (9.6 ± 0.89mm vs. 10.7 ± 1.12mm; P=0.06), in ovulation rate (90.9% vs. 90.9%; P=1) and pregnancy rate (58.8% vs. 62.9%; P=0.79), however, buffalo cows from the ECG treatment have less time between P4 removal and ovulation when compared to EBG buffalos (37.4h vs. 52.8h; P=0.001), respectively. Thus, it was concluded that the implantation of TAI in the floodplain of Amazonas is feasible and the use of EC results in successful rates, similar to EB.(AU)

O objetivo deste trabalho foi determinar a eficiência do cipionato de estradiol (CE) como indutor de ovulação em um protocolo de inseminação artificial de tempo fixo. Para isso, 69 búfalas receberam no D0 um dispositivo intravaginal de progesterona e 2mg de benzoato de estradiol (BE). No D9, o dispositivo intravaginal foi removido e foram aplicados 0,53mg de prostaglandina (PGF2α) e 400UI de gonadotrofina coriônica equina (eCG), para, então, os animais serem divididos em dois grupos: um deles (GCE) recebeu 1mg de CE no D9, e o outro (GBE) recebeu 1mg de BE 24h após. As inseminações ocorreram no D11. A atividade ovariana e o diagnóstico de prenhez foram avaliados por ultrassonografia. Não houve diferença (P>0,05) no diâmetro folicular (9,6 ± 0,89mm vs. 10,7 ± 1,12mm; P=0,06), na taxa de ovulação (90,9% vs. 90,9%; P=1) e na taxa de prenhez (58,8% vs. 62,9%; P=0,79), no entanto búfalas do tratamento GCE apresentaram menor tempo entre a remoção da P4 e a ovulação, quando comparadas com as búfalas do GBE (37,4h vs. 52,8h; P=0,001), respectivamente. A implantação da IATF nas várzeas do Amazonas é viável e a utilização do CE resulta em taxas de sucesso similares ao BE.(AU)

Effect of flaxseed (Linum usitatissimum) and soybean (Glycine max) oils in Egyptian lactating buffalo and cow diets on the milk and soft cheese quality

Produce and compare soft cheese with potential benefits of human health from Egyptian buffalo's and cow's milk was studied. Eight Egyptian lactating buffalos and cows were fed a total mixed ration supplemented with either 0% oil (CD), 2% flaxseed oil (DFO), 2% soybean oil (DSO), or 2% of their mixture (1:1, DFSO) according to a double 4 x 4 Latin Square design. Milk yield was similar between buffalo's diets but was higher in cows fed a DFO, DSO or DFSO resulting in 11.15, 8.21% or 8.97% increases compared with the control diet, respectively. Milk composition was not significantly affected in both buffalos and cows fed diets. The DFO, DSO or DFSO displayed decreased short-chain fatty acids, especially DSO and DFSO (3.73 and 3.33%, respectively) when compared to CD for buffalo milk (6.32%). The DSO and DFSO were more effective for increasing unsaturated fatty acids followed by the DFSO in buffalo's milk fat (42.31 and 41.90 %), whereas DFO and DFSO were more effective in cow's milk fat (39.67 and 39.84%), respectively. DFO, DSO or DFSO had no significant effect on the yield, composition and sensory properties of resultant soft cheese compared to the CD for both lactating cows and buffalos. During storage, a diet rich in unsaturated fatty acids enhances protein proteolysis and antioxidant activity of soft cheese during storage compared to the CD especially for soft cheese produced from buffalo's milk.(AU)

Effect of flaxseed (Linum usitatissimum) and soybean (Glycine max) oils in Egyptian lactating buffalo and cow diets on the milk and soft cheese quality

Produce and compare soft cheese with potential benefits of human health from Egyptian buffalo's and cow's milk was studied. Eight Egyptian lactating buffalos and cows were fed a total mixed ration supplemented with either 0% oil (CD), 2% flaxseed oil (DFO), 2% soybean oil (DSO), or 2% of their mixture (1:1, DFSO) according to a double 4 x 4 Latin Square design. Milk yield was similar between buffalo's diets but was higher in cows fed a DFO, DSO or DFSO resulting in 11.15, 8.21% or 8.97% increases compared with the control diet, respectively. Milk composition was not significantly affected in both buffalos and cows fed diets. The DFO, DSO or DFSO displayed decreased short-chain fatty acids, especially DSO and DFSO (3.73 and 3.33%, respectively) when compared to CD for buffalo milk (6.32%). The DSO and DFSO were more effective for increasing unsaturated fatty acids followed by the DFSO in buffalo's milk fat (42.31 and 41.90 %), whereas DFO and DFSO were more effective in cow's milk fat (39.67 and 39.84%), respectively. DFO, DSO or DFSO had no significant effect on the yield, composition and sensory properties of resultant soft cheese compared to the CD for both lactating cows and buffalos. During storage, a diet rich in unsaturated fatty acids enhances protein proteolysis and antioxidant activity of soft cheese during storage compared to the CD especially for soft cheese produced from buffalo's milk.

Identification of novel single nucleotide polymorphisms in the growth hormone ( GH ) gene in Anatolian water buffalo ( Bubalus bubalis ) populations in Turkey

This study was conducted to investigate the growth hormone (GH; somatotropin-like) gene polymorphisms in 150 water buffalo (Bubalus bubalis) from different regions of Turkey. 404 bp long partial intron 4, exon 5, 3' UTR regions of the GH gene (also called GH/AluI locus) and 347 bp long exon-intron 3 and partial exon 4 regions of the GH gene (also called GH/MspI locus) were amplified, and their PCR products analyzed via DNA sequencing method. Seven genotypes due to twenty single nucleotide polymorphisms (SNP) and one deletion/insertion were identified in a 347 bp long region of the GH/MspI locus. A missense mutation from glycine to glutamate amino acid and four silent mutations in the serine, threonine, and asparagine amino acids were determined in the exon 3 region of the GH gene. Four genotypes due to eight SNP were identified in a 404 bp long region of the GH/AluI locus. A missense mutation from lysine to arginine amino acid and six silent mutations in Leucine, aspartate, histidine, lysine, arginine, and cysteine amino acids were revealed in the exon 5 region of the GH gene. The partial DNA sequence of the GH gene in water buffalos was reported, and these sequences were deposited at the NCBI Genbank database with the accession numbers MN266903-MN266909 and MN530973-MN530976. These SNP may have an effect on economic (such as body composition) and carcass traits, reproduction, and milk yield and content in water buffalo populations and may prove to be useful for water buffalo breeding.(AU)

Rib fracture in buffalo with ruminal cannula

Background: Buffalos are very rustic animals that are adapted to adverse conditions, but the lack of adequate managementcan increase the susceptibility of these animals to several diseases. Rib fracture in this species is usually related to mineraldeficiency and complications resulting from unappropriated management. Rib fractures in ruminants are rarely diagnosedand usually present conservative treatment; however, cases with presence of contaminated wounds require extra attention.The objective of this work was to report an atypical case of open fracture of the last rib in a buffalo with ruminal cannula,describing the case from diagnosis to surgical treatment.Case: An adult female water buffalo (Bubalus bubalis) that was well fed and receiving minerals, with implantation ofa ruminal cannula, presented a lesion around the ruminal cannula, with increasing volume, exudation, and myiasis. Thegeneral parameters and behavior of the animal were apparently normal. The animal presented normal appetite and respiration and good physical condition and nutritional state, with pain reaction to the touch in the region of the lesion andtheir adjacencies; it presented no claudication and reluctance to remain in left lateral decubitus, and no discomfort. Theclinical inspection of the cutaneous lesion showed an oblique-type fracture in the body of the last left rib, with exposedbone in the lumbar region and extremity exposed to the ruminal cannula, already presenting necrosis. The region wascleaned, removing larvae (myiasis) for better exposition of the fracture; in the following day...

Rib fracture in buffalo with ruminal cannula

Background: Buffalos are very rustic animals that are adapted to adverse conditions, but the lack of adequate managementcan increase the susceptibility of these animals to several diseases. Rib fracture in this species is usually related to mineraldeficiency and complications resulting from unappropriated management. Rib fractures in ruminants are rarely diagnosedand usually present conservative treatment; however, cases with presence of contaminated wounds require extra attention.The objective of this work was to report an atypical case of open fracture of the last rib in a buffalo with ruminal cannula,describing the case from diagnosis to surgical treatment.Case: An adult female water buffalo (Bubalus bubalis) that was well fed and receiving minerals, with implantation ofa ruminal cannula, presented a lesion around the ruminal cannula, with increasing volume, exudation, and myiasis. Thegeneral parameters and behavior of the animal were apparently normal. The animal presented normal appetite and respiration and good physical condition and nutritional state, with pain reaction to the touch in the region of the lesion andtheir adjacencies; it presented no claudication and reluctance to remain in left lateral decubitus, and no discomfort. Theclinical inspection of the cutaneous lesion showed an oblique-type fracture in the body of the last left rib, with exposedbone in the lumbar region and extremity exposed to the ruminal cannula, already presenting necrosis. The region wascleaned, removing larvae (myiasis) for better exposition of the fracture; in the following day...(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)

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)