Clinical disorders observed during the first 30 days of life of cloned Zebu calf

Background: The advent of animal cloning from embryos reconstructed with nuclei from cells at different stages of differentiation has been responsible for the appearance of several anomalies in calves, with some alterations diagnosed during pregnancy and others after the birth of cloned animals. Despite efforts over the past five years, the rate of mortality after the birth of cloned calves is still high, at about 50%. Below is reported the research experience gained at the University of São Paulo in the period from 2005 to 2010, related to the birth of cloned calves from Nellore. Review: Research showed that cloned calves present cardiopulmonary disorders characterized by tachycardia with episodes of arrhythmia with bradycardia, increase in the 1st heart sound, presence or absence of heart murmurs in the 1st and 2nd heart sound associated with dyspnea, breathing harsh and crackling dry, moist and crackling. During the first days of life of cloned calves that died, ultrasound examination demonstrated the presence of concentric hypertrophy congenital cardiac and turbulent flow at the foramen of botal towards the right heart to left heart, indicating reversal of blood flow to the pattern observed in fetal life. As a result, mixture of arterial and venous blood occurs, compromising the ability of oxygen to the blood of cloned calves. The occurrence of macrosomia was observed in 20.9% of Nellore calves. It is believed that this syndrome is associated with disorders of carbohydrate metabolism in the placenta and/or fetus, leading to fetal gigantism, which mimicks excessive fetal growth seen in pregnant women with diabetes. We observed the occurrence of moderate to severe normocytic and normochromic anemia, with the anemia gradually appearing from 12 h of life onwards, reaching its maximum intensity at the end of the first week, to start a gradual recovery to normal values from the 15th day of life. The anemia observed in cloned calves was ferropriva, as such animals showed a significant decrease in serum iron levels associated with decreased transferrin saturation index (STI). Umbilical anomalies occurred in most clones. There was an increase in the thickness of the umbilical cord, which hindered its spontaneous rupture at birth. The umbilical arteries did not undergo retraction into the abdominal cavity, leaving them exposed in the remnant of the umbilical cord. In the first three days after birth, strong pulse of these arteries was noted in umbilical cord, making it necessary to use clamps on the arteries in order to prevent bleeding. There were also intra-abdominal hematomas involving the arteries and the urachus. Between 15 and 20 days of life, it was observed the occurrence of alopecia in about 75.0% of the calves, whose origin may be related to disturbances in the synthesis and absorption of vitamins, since the supplementation of calves with ADE vitamin complex decreased symptoms. Conclusion: The results confirm the occurrence of serious cardiopulmonary disorders characterized by tachycardia, hyperphonesis, presence of heart murmurs in the 1st and 2nd heart sounds, episodes of arrhythmia and bradycardia associated with dyspnea, rales, and coarse breath. As a result of the non-closure of the foramen of Botal and the Ductus Arteriosus, arterial and venous blood a mix, compromising the oxygen carrying capacity in the blood of cloned calves. There was also the occurrence of macrosomia, hypoglycemia, hypothermia, abnormal umbilical structures, anemia and alopecia

Teores de proteína, gordura e sólidos totais no leite de vacas da raça Jersey criadas no Estado de São Paulo durante o primeiro mês de lactação / Proteins, fats and total solids content in milk from Jersey cows raised in São Paulo State during the first month of lactation

Com a finalidade de estabelecer os valores de referência dos teores de proteína, gordura e sólidos totais do leite de bovinos da raça Jersey,criados no Estado de São Paulo, durante o primeiro mês de lactação,bem como avaliar a influência da fase colostral, examinaram-se 418 amostras de leite, provenientes de quartos mamários sadios e sem crescimento bacteriano. O leite foi colhido assepticamente antes da ordenha e os valores de proteína, gordura e sólidos totais foram determinados por radiação infra vermelho. Os teores de proteína na secreção láctea diminuíram abruptamente do 1º para o 2º dia de lactação,enquanto os teores de sólidos totais diminuíram abruptamente nas primeiras 24 horas após o parto. Após oscilaram muito nos primeiros três dias de lactação, observou-se que os valores de gordura diminuíam atingindo os valores mínimos entre o 15º e 30º dia de lactação. Sugere-se nas primeiras 24 horas de lactação (período colostral) a adoção dos seguintes valores de referência: proteína - entre 7,35 e 11,45 g/dL;gordura 0,98 e 2,00 g/dL; e sólidos totais - entre 12,39 e 19,45 g/dL(AU)

With the aim to establish reference values of the protein, fat and total solids of milk from Jersey cows, raised in the State of São Paulo, during the first month of lactation, as well as to evaluate the influence of the colostral phase, 418 milk samples, obtained from healthy mammary glands without bacterial growth, were analized. Milk samples were aseptically collected before milking and protein, fat and total solids determined by infrared radiation. The milk protein level decrease from first to second day of lactation, while milk total solids level decreased during the first 24 hours after parturition. Milk fat level oscillated in the first 3 days of lactation and then decreased to the minimum values between 15-30 days of lactation. We suggest, forthe first 24 hours of lactation (colostral phase) the use of the following reference values: protein- between 7,35 and 11,45 g/dl; fat- between 0,98 and 2,00 g/dl ; total solids between 12,39 and 19,45 g/dl(AU)

O uso da cardiotocografia como método de diagnóstico da ocorrência de sofrimento fetal (hipóxia fetal) durante a vida intrauterina de fetos da raça Nelore originados por meio da técnica de transferência nuclear de células somáticas adultas - Clonagem / The use of cardiotocography as a method of diagnosis of the occurrence of fetal distress (fetal hypoxia) during intrauterine life in fetuses of Nellore generated through the technique of somatic adult cell nuclear transfer - Cloning

A presente pesquisa teve a finalidade de padronizar a técnica de cardiotocografia a ser utilizada para avaliar a existência de hipóxia/sofrimento fetal durante a vida intrauterina, procurando estabelecer a partir do 7º mês da gestação os padrões de normalidade da frequência cardíaca fetal para fetos sadios e oriundos de inseminação artificial, bem como avaliar a influência da clonagem nos parâmetros obtidos no exame de cardiotocografia. Foram acompanhadas as gestações de 14 vacas, sendo os animais divididos em três grupos experimentais: Grupo de Clones Mortos composto de 5 vacas gestantes nas quais acompanhou-se a carditocografia de fetos gerados pela técnica de clonagem e nos quais a morte do bezerro ocorreu nas primeiras 36 horas de vida ; Grupo de Clones Vivos composto de 4 vacas gestantes nas quais acompanhou-se a carditocografia de fetos gerados pela técnica de clonagem e que permaneceram vivos após o nascimento; Grupo Controle - composto de 5 vacas gestantes nas quais acompanhou-se a carditocografia de fetos gerados por inseminação artificial e que deram origem a bezerros sadios. Os animais foram acompanhados por exames periódicos de cardiotocografia realizados nos seguintes momentos: 90, 60, 30, 15, 7, 6, 5, 4, 3, 2, 1 dia antes do parto e no dia do parto. Para a realização dos exames foi utilizado o aparelho de cardiotocografia da marca Philips modelo Avalon FM30,no local onde as vacas se encontravam. Os exames tiveram uma duração de 20 a 40 minutos de traçado, sendo que o transdutor de ultra-som era acomodado de forma a obter o melhor sinal cardíaco fetal. Imediatamente após o nascimento, realizou-se a avaliação da viabilidade de bezerros recém-nascidos pelo método APGAR e a observação de bezerros tingidos de amarelo em decorrência da presença de mecônio nas secreções uterinas.Através dos traçados obtidos com a cardiotocografia foram analisados os parâmetros da cardiotocografia: linha de base, acelerações transitórias, acelerações prolongadas, variabilidade, taquicardia, bradicardia e desacelerações, sendo ao final calculado o índice cardiotocométrico. Nos animais classificados como hipoativo ou inativo aplicava-se estimulação por palpação retal com beliscamento do espaço interdigital do feto com o objetivo de modificar o comportamento fetal e evitar resultados falso positivos.O exame de cardiotocografia mostrou ser eficaz na avaliação da vitalidade fetal na espécie bovina. Os valores médios e o desvio padrão da frequência cardíaca fetal basal (linha de base) em fetos hígidos foi igual a 116,4 ± 9,07 bpm. Considerou-se taquicardia uma frequência cardíaca fetal igual ou maior do que 145 bpm e bradicardia uma frequência cardíaca igual ou menor do que 90 bpm. Considerou-se normal para bovinos uma variabilidade com valores situados entre 5 e 15 bpm. Verificou-se que a presença de acelerações transitórias são indicativas de bem estar fetal, sendo que o número de acelerações transitórios consideradas como fisiológicas varia entre 1 e 5. Desacelerações são indicativos da existência de sofrimento fetal, não sendo observadas em animais hígidos. No grupo de clones mortos, existem modificações da carditocografia compatíveis com a ocorrência de sofrimento fetal/ hipóxia intrauterina. Os fetos do grupo de clones mortos apresentaram com maior frequência momentos de hipoatividade fetal do que os fetos do grupo de clones vivos e fetos do grupo controle e os fetos do grupo de clone mortos foram classificados com maior frequência como não reativos ao estímulo de beliscamento das extremidades do que os fetos do grupo controle. Os fetos do grupo de clones mortos apresentaram com mais frequência bradicardia (11,8%) do que a observada no grupo controle (0,00 %). Observou-se que no grupo de clones mortos (1,4 ± 1,6) o número de acelerações transitórias foi significativamente menor do que o observado no grupo de clones vivos (3,50 ± 2,70) e no grupo controle (2,70 ± 1,80)

This research had the purpose to standardize the technique of cardiotocography to be used to assess the presence of hypoxia / fetal distress during intrauterine life, trying to establish from the 7th month of pregnancy the normal range of fetal heart rate in healthy fetuses and from artificial insemination and cloning to evaluate the influence of the parameters obtained in the examination of cardiotocography. Were followed the pregnancies of 14 cows and animals are divided into three experimental groups: Dead Clones Group - composed of 5 pregnant cows which was accompanied the cardiotocography of fetuses generated by the technique of cloning, in which the death occurred in the calf first 36 hours of life; Living Clones Group - composed of 4 pregnant cows which is accompanied cardiotocography of fetuses generated by the technique of cloning and remained alive after the birth, control group - composed of 5 pregnant cows in which follow - the cardiotocography of fetuses generated by artificial insemination and gave rise to healthy calves. The animals were monitored by periodic assessment of cardiotocography performed in the following periods: 90, 60, 30, 15, 7, 6, 5, 4, 3, 2, 1 day before parturition and day of birth. For the examinations we used the apparatus of cardiotocography Brand Philips Avalon FM30 model, where the cows were. The tests had a duration of 20 to 40 minutes of track, and the ultrasound transducer was adjusted to obtain the best fetal cardiac signal. Immediately after birth, was held to assess the viability of newborn calves by the method APGAR and the observation of colored yellow calves due to the presence of meconium in the uterine secretions. Through cardiotocography tracings were analyzed with the parameters of cardiotocography : the baseline, acceleration transient, prolonged accelerations, variability, tachycardia, bradycardia, and deceleration modes, and the final calculated the index cardiotocométrico. In the animals classified as inactive or hypoactive stimulation was applied by rectal palpation of the interdigital space with pinch of the fetus with order to modify the behavior and prevent fetal false positives. An examination of cardiotocography has proven effective in evaluation of fetal vitality in the bovine species. The mean values and standard deviation of the baseline fetal heart rate (from baseline) in healthy fetuses was equal to 116.4 ± 9.07 bpm. Tachycardia was considered a fetal heart rate equal to or greater than 145 bpm bradycardia and a heart rate less than or equal to 90 bpm. It was considered normal for cattle variability with values between 5 and 15 bpm. It was found that the presence of transient accelerations are indicative of fetal well-being, and the number of transient accelerations considered physiological ranges between 1 and 5. Deceleration is indicative of the existence of fetal distress and is not observed in healthy animals. In the group of clones dead, there are changes in cardiotocography consistent with the occurrence of fetal distress / intrauterine hypoxia. The fetuses of the group of clones had killed more often moments of fetal hypoactivity than the fetuses of the group of clones live fetuses and the control group and the group of fetuses dead clone were classified more frequently as non-reactive to stimulation of the pinch ends of the fetuses in the control group. The fetuses of the group of dead clones showed bradycardia more often (11.8%) than that observed in the control group (0.00%). It was observed that the group of dead clones (1.4 ± 1.6) the number of transient acceleration was significantly lower than that observed in the group of live clones (3.50 ± 2.70) and the control group (2 , 70 ± 1.80)

Ocorrência de resíduos de antibióticos no leite, em fazendas produtoras e no leite pronto para consumo / Occurrence of antibiotic residues in milk, producing farms and ready milk for consumption

Um ponto fundamental na qualidade do leite é a ausência de contaminantes, como, por exemplo, resíduos de antimicrobianos. A presença de resíduos de antibióticos no leite resulta da aplicação de diferentes substâncias antimicrobianas no efetivo leiteiro, para prevenção ou tratamento de doenças, com destaque para infecções da glândula mamária e as doenças do trato reprodutivo. As razões para se fazer o controle de resíduos de antibióticos no leite incluem a possibilidade desses resíduos causarem reações alérgicas nos consumidores e provocarem o surgimento de resistência bacteriana, além de ser também um problema econômico, interferindo nas culturas lácteas utilizadas na fabricação de derivados. Os limites máximos permitidos (LMP) para resíduos de drogas para uso veterinário em alimentos são determinados pelo Codex Alimentarius, da FAO (Food and Agriculture Organization, as ONU), e da Organização Mundial da Saúde (OMS). Para a detecção de antimicrobianos no leite, testes microbiológicos, rápidos e de grupos específicos são comumente aplicados, que se baseiam na inibição do crescimento bacteriano, em reações imunológicas ou enzimáticas e na separação, identificação e quantificação do antimicrobiano por meio de técnicas de cromatografia gasosa ou líquida. Foram utilizadas 20 propriedades produtoras e 30 amostras de leite pronto para consumo no Estado de São Paulo. Para a detecção dos resíduos de antibiótico foi utilizado o teste microbiológico comercial Delvotest® SP para betalactâmicos, cujo princípio é a inibição do crescimento microbiano.(AU)

An important point about milk quality is a free contamination, like a free antibiotics residue. The presence of antibiotics residue in the milk provide of the use of differents kinds of antibiotics on the dairy herd, to prevent ortreatment of diseases, that we can highlights mammary infection and reproductive tract. Many reasons to control residue of antibiotics in milk, includes human allergic reactions, and increase of bacterial resistence, besides a big economic problem to make milk products, that have an interference at bacterial cultures. PML (Permised Maximum Limited) to veterinary drugs residues at food, is determined by Codex Alimentarius, a FAO(Food and Agriculture Organization), and WHO (World Healf Organization).To detect antibiotics in milk, microbiologics test,fast and specific, that is based on bacterial innibition growth; at immunologic or enzimatic reation and duringg separation identification, and quantification process of the antibiotics by gas or liquid cromatografy. On thisproject, we used 20 dairy properties and 30 milk prepared for use samples at São Paulo State. To detect antibiotc residue, we used a commercial microbiologic test (Oelvotest® SP) to detect betalactamics bacterial, that inhibit thebacterial growth. (AU)