Trials with the Haemonchus vaccine, Barbervax®, in ewes and lambs in a tropical environment: Nutrient supplementation improves protection in periparturient ewes.

Haemonchus contortus is an economic problem in sheep farms worldwide, mainly in the tropics and subtropics. A vaccine against haemonchosis, called Barbervax®, was evaluated in ewes under two nutritional status, naturally infected with gastrointestinal nematodes. Ewes were divided into four groups: Supplemented Diet - Vaccine; Supplemented Diet - No vaccine; Basal Diet - Vaccine and Basal Diet - No vaccine. Their lambs were divided in Vaccinated and No vaccine. Ewes were immunised six times starting about 1 month of pregnancy with the first three doses at 3 week intervals and the last three shots at 4 week intervals. Supplemented ewes had higher body weight, body score and packed cell volume compared with those fed a basal diet. Both groups of vaccinated ewes showed a similar response in circulating anti-vaccine antibodies but the vaccine had no discernible effect on either body weight, body score and packed cell volume. There was a marked group difference in the number of ewes that received precautionary treatments with anthelmintic. All 14 Basal Diet - No vaccine ewes required treatment. In contrast only 7 ewes, in the Supplemented Diet - Vaccine group required anthelmintic treatment. In the Basal Diet - Vaccine and in the Supplemented Diet - No Vaccine groups, 12 and 13 ewes needed anthelmintic treatment, respectively. Vaccinated lambs showed much higher antibody titres resulting in 80% less Haemonchus spp. egg counts comparing with no vaccine lambs. Taken together these results clearly suggest that in pregnant and lactating ewes a combined protective effect between vaccination and improved nutrition resulted in fewer precautionary anthelmintic treatments. Thus, it was possible to achieve a more sustainable level of control of the haemonchosis, less dependent on anthelmintic drugs.

Análise da variabilidade da frequência cardíaca em cordeiros da raça Bergamácia, do nascimento aos 35 dias de idade / Analysis of heart rate variability in Bergamasca newborn lambs from birth at 35th days of age

A atividade do sistema nervoso autônomo sobre o coração pode ser verificada pela variabilidade da frequência cardíaca (VFC), método que quantifica e assim infere sobre a modulação autonômica cardíaca, refletindo o funcionamento do sistema nervoso autônomo. Durante o período neonatal, diferenças significativas no que se refere à maturação autonômica são descritas em diversas espécies. Embora a espécie ovina venha sendo utilizada como modelo experimental em diversos protocolos em neonatologia e cardiologia em humanos, estudos descritivos sobre a VFC utilizando animais saudáveis e não sedados são escassos na literatura. O objetivo do estudo foi descrever a VFC em cordeiros da raça Bergamácia durante os primeiros 35 dias de idade. Foram realizados exames eletrocardiográficos em 20 cordeiros da raça Bergamácia a partir do nascimento e semanalmente até o 35° dia de idade. A VFC foi analisada pelo intervalo RR normal (batimentos sinusais), pela frequência cardíaca, pelo índice de tônus vasovagal (iTVV), pela raiz quadrada da média do quadrado das diferenças entre intervalos RR normais adjacentes (RMSSD) e pela raiz quadrada da somatória do quadrado das diferenças dos valores individuais em relação ao valor médio, dividido pelo número de iR-R em 90 segundos - VFC global (RMSM). Todos os parâmetros se alteraram ao longo das semanas. A frequência cardíaca média elevou-se entre o nascimento e os primeiros sete dias de idade, com decréscimo nas quatro semanas subsequentes, sendo o menor valor encontrado aos 35 dias de idade (145,63±37,80bpm). Entre 21, 28 e 35 dias de idade, o iTVV elevou-se significativamente, o RMSM a partir do 28º dia, e, aos 35 dias, o RMSSD, reflexo da ativação parassimpática, exibiu diferenças em relação aos momentos subsequentes. O início da predominância parassimpática, refletida nos índices da VFC, particularmente o iTVV, ocorre aos 21 dias de idade. A partir dos 35 dias de idade, os índices RMSM e RMSSD podem ser utilizados como marcadores fidedignos das mudanças nos efeitos simpático e parassimpático sobre o coração de cordeiros.(AU)

Autonomic nervous system activity can be checked by heart rate variability (HRV), method that quantifies and thus infers on cardiac autonomic modulation, reflecting the functioning of the autonomic nervous system. During the neonatal period significant differences as regards the autonomic maturation, are described in various species. Although the sheep come being used as experimental model of several protocols in neonatology and cardiology in humans, descriptive studies about the VFC using healthy animals and not sedated are scarce in the literature. The objective of this study was to describe the HRV in the Bergamasca race lambs during the first 35 days of age. Electrocardiographic examinations were performed in 20 lambs of the Bergamasca race from birth, in the7th, 14th, 21st, 28th, and 35th days of age. The HRV was analyzed by normal RR interval (sinus rate), heart rate, index of vagal tone, and the cardiovascular manifestations (square root of the average of the square of the differences between normal RR intervals adjacent). All parameters have changed over the weeks. The average heart rate increased between the birth and the first seven days of age, with decrease in the subsequent four weeks, being the smallest value found at 35 days of age (145.63±37.80bpm). Between 21, 28 and 35 days of age, the iTVV increased significantly, the RMSM from the 28 day, and 35 days, the cardiovascular manifestations, reflex parasympathetic activation, exhibited differences in relation to subsequent times. The beginning of parasympathetic dominance, reflected in the HRV indices, particularly the iTVV, occurs at 21 days of age. From the 35 days of age, the RMSM indices and RMSSD can be used as reliable markers of change in sympathetic and parasympathetic effect on the heart of lambs.(AU)

Influence of feeding differentiation on the age at onset of puberty in Brazilian Bergamasca dairy ewe lambs / Influência da diferenciação alimentar na idade à puberdade em cordeiras da raça Bergamácia Brasileira

The objective of the study was to evaluate body growth and age at onset of puberty on lambs fed two specific diets for low and high growth rates. A herd of 20 Brazilian Bergamasca lambs was divided in two groups (n= 10) and kept confined throughout the experimental period, two animals of the same treatment/pen. Two phases were established: Phase 1, from 90 days of age until the onset of puberty; and Phase 2, from puberty onset up to 1 year old. For Phase 1, two distinct diets were formulated, being: Treatment A, which was formulated to obtain an average daily gain of approximately 150g; and Treatment B, for an average daily gain of about 250g. In Phase 2, a balanced, equal diet was provided to both groups. Every 14 days, the animals were weighed and given average daily gain, average daily dry matter intake and body condition score. From the 5th month of age on, in each group, a vasectomized male was used to detect estrus, establishing age at puberty onset and estrus interval for each lamb. Blood samples were collected every 28 days to determine plasma growth hormone concentration. Treatment B lambs gained more weight and had higher body condition score (P<0.05) and there was no difference for age at puberty onset and plasma growth hormone levels (P>0.05) between treatments. It was found that both treatments showed satisfactory performances. Thus, treatment A may be indicated as a reasonable feeding system to achieve positive responses on confined ewe lambs during growth phase.

O objetivo do estudo foi avaliar o crescimento corporal e idade de início da puberdade em cordeiras sob duas dietas específicas para taxas de crescimento baixas e altas. Um rebanho de 20 cordeiras Bergamácia Brasileira foi dividido em dois grupos (n= 10) e mantido confinado em todo período experimental, sendo dispostos dois animais do mesmo tratamento/baia. Duas fases foram estabelecidas: Fase 1, a partir de 90 dias de idade até o início da puberdade, e Fase 2, a partir do início da puberdade até 1 ano de idade. Para a fase 1, duas dietas distintas foram formuladas, sendo: Tratamento A, formulada para obter um ganho de peso diário de aproximadamente 150g; e Tratamento B, para um ganho de peso médio diário de cerca de 250g. Na Fase 2, uma dieta equilibrada idêntica foi fornecida para ambos os grupos. A cada 14 dias, os animais foram pesados e calcularam-se o ganho médio diário, o consumo médio diário de matéria seca e escore de condição corporal. A partir do quinto mês de idade, em cada grupo, um macho vasectomizado foi usado para detectar estro, estabelecendo a idade de início da puberdade e estro para cada cordeira. As amostras de sangue foram coletadas a cada 28 dias para determinar a concentração plasmática de hormônio do crescimento. Cordeiras do tratamento B ganharam mais peso e tiveram maior escore de condição corporal (P<0,05), mas não houve diferença de idade para o início da puberdade e para os níveis plasmáticos de hormônio do crescimento (P>0,05). Verificou-se que os tratamentos A e B apresentaram desempenhos satisfatórios. Assim, o tratamento A pode ser indicado como um sistema de alimentação para alcançar respostas positivas em cordeiras submetidas ao confinamento durante a fase de crescimento.

Attempts to vaccinate ewes and their lambs against natural infection with Haemonchus contortus in a tropical environment.

A vaccine containing integral membrane glycoproteins from the intestine of Haemonchus contortus was evaluated in three groups of grazing sheep each containing 13 ewes and their 16 lambs naturally infected with gastrointestinal nematodes. Two groups were vaccinated with either 5 or 50µg of the antigen per immunisation, while the third, the control group, received adjuvant alone. The sheep were immunised six times at 3week intervals, partly because the vaccine antigens are hidden and thus no immunological boost would be delivered by subsequent infection and partly because the level of Haemonchus spp. challenge was expected to be high. The vaccinated ewes, first immunised approximately 1month before lambing, showed a circulating antibody response but no signs of reduced anaemia or Haemonchus spp. egg counts, compared with control ewes. Several ewes with severe haemonchosis in all three groups had to be given precautionary treatment with anthelmintic drugs. In contrast, vaccinating their lambs with either 5 or 50µg of the antigen per immunisation resulted in 10 fold higher antibody titres. In the case of the lower antigen dose this was associated with significantly less anaemia, 72% reduction in the overall number of Haemonchus spp. eggs produced and significantly fewer worms compared with control lambs. It is hypothesised that the heavily pregnant or lactating ewes did not have sufficient physiological reserves to mount a protective response following vaccination in the tropical weather and high challenge conditions that prevailed. Nevertheless, the vaccine could afford useful protection for lambs against H. contortus.

Pregnancy rate and birth rate of calves from a large-scale IVF program using reverse-sorted semen in Bos indicus, Bos indicus-taurus, and Bos taurus cattle.

Obtaining sexed sperm from previously frozen doses (reverse-sorted semen [RSS]) provides an important advantage because of the possibility of using the semen of bulls with desired genetic attributes that have died or have become infertile but from whom frozen semen is available. We report the efficiency of RSS on the pregnancy rate and birth rate of calves in a large-scale program using ovum pick-up and in vitro embryo production (IVEP) from Bos indicus, Bos indicus-taurus, and Bos taurus cattle. From 645 ovum pick-up procedures (Holstein, Gir, and Nelore), 9438 viable oocytes were recovered. A dose of frozen semen (Holstein, Nelore, Brahman, Gir, and Braford) was thawed, and the sperm were sex-sorted and cooled for use in IVF. Additionally, IVF with sperm from three Holstein bulls with freeze-thawed, sex-sorted (RSS) or sex-sorted, freeze-thawed (control) was tested. A total of 2729 embryos were produced, exhibiting a mean blastocyst rate of 29%. Heifers and cows selected for adequate body condition, estrus, and health received 2404 embryos, and 60 days later, a 41% average pregnancy rate was observed. A total of 966 calves were born, and 910 were of a predetermined sex, with an average of 94% accuracy in determining the sex. Despite the lower blastocyst rate with freeze-thawed, sex-sorted semen compared with sex-sorted semen, (P < 0.05), the pregnancy rate (bull I, 45% vs. 40%; II, 35% vs. 50%; and III, 47% vs. 48% for RSS and control, respectively; P > 0.05) and sex-sorted efficiency (bull I, 93% vs. 98%; II, 96% vs. 94%; and III, 96% vs. 97% for RSS and control, respectively; P > 0.05) were similar for each of the three bulls regardless of the sperm type used in the IVF. The sexing of previously frozen semen, associated with IVEP, produces viable embryos with a pregnancy rate of up to 40%, and calves of the desired sex are born even if the paternal bull has acquired some infertility, died, or is located a long distance from the sexing laboratory. Furthermore, these data show the feasibility of the process even when used in a large-scale IVEP program.

High tumor necrosis factor-α/interleukin-10 ratio is associated with hepatocellular carcinoma in patients with chronic hepatitis C.

Hepatitis C virus (HCV) is the main cause of chronic liver disease, cirrhosis and hepatocellular carcinoma (HCC) worldwide. The risk for the development of HCC increases with the severity of liver inflammation and fibrosis. The hepatic inflammation caused by HCV involves host regulatory immune response, which is mediated by cytokines with anti-viral role upon the interaction of viral polypeptides with innate and adaptive immunity. Two cytokines; tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) play key roles in the regulation of cellular immune response in HCV infection. The aim of the present study was to determine the levels of IL-10 and TNF-α, as well as the ratio of TNF-α and IL-10 serum levels in patients with HCV and HCC caused by HCV (HCC-HCV). The study included 173 patients with chronic HCV. TNF-α and IL-10 serum levels were measured by ELISA (R&D Systems, Inc.). In the present study, 54 patients presented liver mild fibrosis, 68 had severe fibrosis and 51 patients had HCC. After adjustment in the multivariate regression analysis, the following variables remained significantly associated with HCC-HCV occurrence: diabetes (p=0.012 OR 10.44 CI 1.66-65.60), IL-10 lower levels (p<0.0001 OR 0.83 CI 0.78-0.89) and TNF-α higher levels (p<0.0001 OR 1.19 CI 1.11-1.28). Individuals with HCC presented higher TNF-α/IL-10 ratio than those with fibrosis grade F4, F3 or F0+F1+F2 (p=0.0003, p<0.0001, p<0.0001, respectively). Patients with HCC were associated to higher index TNF-α/IL-10 ratio, suggesting that the unbalanced production of these cytokines may represent progression to the liver disease severity in HCV infected patients.

Effects of hepatitis C virus on cardiovascular risk in infected patients: a comparative study.

The role of hepatitis C virus (HCV) in the pathogenesis of atherosclerosis and cardiovascular events is unclear. The aim of this study was to evaluate the direct effect of HCV on cardiovascular risk and correlate it with pro and anti-inflammatory cytokines in patients with HCV. HCV monoinfected patients, genotype 1, naive, non-obese (BMI<30) and non-diabetics were included and compared to controls (blood donors). Patients with prior diagnosis of cardiovascular diseases, hypertension, chronic renal failure, cancer and chronic use of lipid-lowering drugs or immunosuppressants were excluded. Age, BMI, systolic blood pressure (SBP) and diastolic (DBP), fasting glucose and lipid levels were determined. Serum cytokines (IL-6, IL-10 and TNF-α) and Framingham score were also evaluated. 62 HCV patients, 34 (54.8%) were males and none of them was smoking. The Framingham scores (median and 25th and 75th percentiles) were 12% (6.5-14%), showing an intermediate cardiovascular risk in patients with HCV. There was significant direct correlation between Framingham and total cholesterol (p=0.043) and DBP (p=0.007). HDL-C (p=0.002) was inversely correlated with the Framingham score. HCV patients had higher levels of proinflammatory cytokines (IL-6 and TNF-α) compared to controls (p<0.0001) and the relation of proinflammatory/anti-inflammatory TNF-α/IL10 and IL-6/IL10 were higher in HCV patients (p<0.01). The Framingham score was directly correlated to IL-6 and TNF-α, but differences were not statistically significant. Patients with HCV monoinfected, nonobese, naïve and non diabetic have an intermediate cardiovascular risk, as measured by the Framingham score and high levels of proinflammatory cytokines (IL-6 and TNF).

MTP -493G/T gene polymorphism is associated with steatosis in hepatitis C-infected patients

The reduction of hepatic microsomal transfer protein (MTP) activity results in fatty liver, worsening hepatic steatosis and fibrosis in chronic hepatitis C (CHC). The G allele of the MTP gene promoter, -493G/T, has been associated with lower transcriptional activity than the T allele. We investigated this association with metabolic and histological variables in patients with CHC. A total of 174 untreated patients with CHC were genotyped for MTP -493G/T by direct sequencing using PCR. All patients were negative for markers of Wilson’s disease, hemochromatosis and autoimmune diseases and had current and past daily alcohol intake lower than 100 g/week. The sample distribution was in Hardy-Weinberg equilibrium. Among subjects with genotype 1, 56.8 percent of the patients with fibrosis grade 3+4 presented at least one G allele versus 34.3 percent of the patients with fibrosis grade 1+2 (OR = 1.8; 95 percentCI = 1.3-2.3). Logistic regression analysis with steatosis as the dependent variable identified genotypes GG+GT as independent protective factors against steatosis (OR = 0.4, 95 percentCI = 0.2-0.8; P = 0.01). The results suggest that the presence of the G allele of MTP -493G/T associated with lower hepatic MTP expression protects against steatosis in our CHC patients.

MTP -493G/T gene polymorphism is associated with steatosis in hepatitis C-infected patients.

The reduction of hepatic microsomal transfer protein (MTP) activity results in fatty liver, worsening hepatic steatosis and fibrosis in chronic hepatitis C (CHC). The G allele of the MTP gene promoter, -493G/T, has been associated with lower transcriptional activity than the T allele. We investigated this association with metabolic and histological variables in patients with CHC. A total of 174 untreated patients with CHC were genotyped for MTP -493G/T by direct sequencing using PCR. All patients were negative for markers of Wilson's disease, hemochromatosis and autoimmune diseases and had current and past daily alcohol intake lower than 100 g/week. The sample distribution was in Hardy-Weinberg equilibrium. Among subjects with genotype 1, 56.8% of the patients with fibrosis grade 3+4 presented at least one G allele versus 34.3% of the patients with fibrosis grade 1+2 (OR = 1.8; 95%CI = 1.3-2.3). Logistic regression analysis with steatosis as the dependent variable identified genotypes GG+GT as independent protective factors against steatosis (OR = 0.4, 95%CI = 0.2-0.8; P = 0.01). The results suggest that the presence of the G allele of MTP -493G/T associated with lower hepatic MTP expression protects against steatosis in our CHC patients.

Composição tecidual e qualidade da gordura na carne de cordeiros castrados e não castrados confinados sob dois fotoperíodos / Tissue composition and fat quality of meat of intact or castrated male lambs confined under two photoperiods

Avaliaram-se os efeitos do fotoperíodo e da castração sobre a composição dos tecidos da paleta e características de qualidade da gordura do lombo e da paleta, de 20 cordeiros mestiços Ideal, distribuídos em esquema fatorial 2 x 2 (dois fotoperíodos - curto (FC), com 12 horas de luz, e longo (FL), com 18 horas de luz, e duas condições sexuais - não castrados (NC) e castrados (C)), com cinco repetições. Os animais foram abatidos aos 37kg de peso corporal. Maior quantidade de gordura total ocorreu nos cordeiros C e mais tecido conjuntivo nos animais NC. A castração influenciou o resíduo mineral fixo (RMF), o extrato etéreo (EE) e a proteína da gordura subcutânea. O efeito da interação entre fotoperíodo longo e castração resultou em aumento do teor de umidade na gordura intermuscular da paleta. A castração elevou o teor de EE e diminuiu o percentual de RMF. Não foi evidenciado efeito do fotoperíodo no EE dos músculos da paleta, e os animais castrados apresentaram gordura intramuscular mais elevada. Os níveis de colesterol da paleta foram mais elevados que os do lombo. Na carne de animais C, verificou-se maior quantidade de ácidos graxos saturados.

The effects of photoperiod and castration on tissue muscle composition, fat physical-chemical composition, and cholesterol was determined for two muscles of 20 Ideal crossbred lambs. The animals were divided into four treatments: five intact males and five castrate during a short photoperiod of 12 light hours, and five intact males and five castrated during a long photoperiod of 18 light hours. The animals were allocated in individual pens, in two identical rooms, with light intensity of 300 lux, (intact and castrated animals x short and long photoperiod). The animals were slaughtered as they reached 37kg of body weight. Castrated lambs showed a significantly higher amount of total fat tissues while intact animals showed higher connective tissue for the shoulder tissue composition. Effect of castration on ash, lipid, and protein was observed in the subcutaneous fat tissue. Interaction effect between LF and castration promoted in shoulder fat intermuscular tissue increase of moisture content while the castration increased the lipid content and decreased the ash. Castrated animals presented higher intramuscular fat. Castrated lambs showed cholesterol contents in shoulder muscles larger than in the Longissimus lumborum muscle. The castration promoted larger amount of saturated fatty acids.

Desempenho de cordeiros Corriedale, puros e mestiços, terminados em pastagem e em confinamento / Performance of Corriedale and crossbred lambs raised on pasture and on drylot

O experimento teve por objetivo avaliar o desempenho de cordeiros, do nascimento ao abate com 30-32kg de peso vivo na origem, usando-se 23 cordeiros Corriedale (C), 25 Bergamácia x Corriedale (BC) e 17 Hampshire Down x Corriedale (HC), terminados em pastagem de coast cross (Cynodon dactylon) ou em confinamento. O peso ao nascimento e o ganho de peso médio diário do nascimento ao desmame näo foram influenciados pelo grupo genético (P>0,05). O ganho de peso médio diário do desmame ao abate (GPDA) de 0,144kg dos cordeiros confinados foi superior aos 0,106kg dos cordeiros em pastagem (P<0,05). O GPDA dos cordeiros BC (0,136kg) e HC (0,130kg) foram maiores (P<0,05) que o dos C (0,106kg). O sistema de terminaçäo e de cruzamento (P<0,05) influiu no ganho de peso do nascimento ao abate (GPNA), com maiores ganhos observados para cordeiros confinados (0,137kg) e cruzados (0,132kg). Os cordeiros confinados atingiram o peso de abate (219 dias) mais cedo (P<0,05) que os da pastagem (258 dias). Os cordeiros terminados em confinamento, no dia do abate, apresentaram menor perda de peso da origem ao abate (3,16 por cento), comparados aos da pastagem (5,17 por cento) (P<0,05). Os resultados indicam que pode-se recomendar o cruzamento de ovelhas Corriedale com carneiros das raças citadas, bem como a terminaçäo dos cordeiros em confinamento, desde que se tenha preocupaçäo de avaliar os custos de produçäo deste sistema