Changes in expression levels of Nod-like receptors in the spleen of ewes

Nucleotide-binding oligomerization domain receptors (NOD-like receptors, NLRs) have critical effects on interfaces of the immune and reproductive systems, and the spleen plays a key role in both innate and adaptive immune functions. It is hypothesized that NLR family participates in maternal splenic immune regulation during early pregnancy in sheep. In this study, maternal spleens were collected on day 16 of the estrous cycle, and days 13, 16 and 25 of gestation (n = 6 for each group) in ewes. Expression of NLR family, including NOD1, NOD2, class II transactivator (CIITA), NLR family apoptosis inhibitory protein (NAIP), nucleotide-binding oligomerization domain, Leucine rich repeat and Pyrin domain containing 1 (NLRP1), NLRP3 and NLRP7, was analyzed using quantitative real-time PCR, Western blot and immunohistochemistry analysis. The results revealed that expression levels of NOD1, NOD2, CIITA and NLRP3 were downregulated at days 13 and 16 of pregnancy, but expression of NLRP3 was increased at day 25 of pregnancy. In addition, expression values of NAIP and NLRP7 mRNA and proteins were improved at days 16 and 25 of pregnancy, and NLRP1 was peaked at days 13 and 16 of pregnancy in the maternal spleen. Furthermore, NOD2 and NLRP7 proteins were limited to the capsule, trabeculae and splenic cords. In summary, early pregnancy changes expression of NLR family in the maternal spleen, which may be related with the maternal splenic immunomodulation during early pregnancy in sheep.(AU)

Antimicrobial resistance of Shiga toxin­producing Escherichia coliisolated from sheep / Escherichia coli produtora de toxina Shiga resistentes a antimicrobianos isoladas de ovinos

The presence of Shiga toxin-producing Escherichia coli (STEC) and resistance to beta-lactams in healthy sheep represents a potential public health risk. This study aimed to characterize STEC isolates in sheep feces for toxin production and resistance to beta-lactam antibiotics. In the present study, among the 40 isolates, we found a predominance of subtype Stx1 (22/40), followed by subtype Stx1 + Stx2 (11/40), while the less prevalent group was Stx2 (7/40). Also, we found phenotypical resistance to beta-lactam antibiotics in 50% (20/40) of the strains analyzed, forming two groups, one with resistant isolates and the other with non-resistant isolates. The cytotoxicity of the isolates did not vary among the groups. In addition to having this characteristic, some multiresistant isolates produced significant amounts of toxins. This leads to the conclusion that the mechanisms of antimicrobial resistance via beta-lactamases are present in sheep STEC and that the cytotoxicity of those isolates is variable regarding such resistance.(AU)

A presença de Escherichia coli produtora de toxina Shiga (STEC) e resistente a beta-lactâmicos em ovinos saudáveis, representa um risco potencial para a saúde pública. O objetivo deste estudo foi caracterizar isolados STEC presentes em fezes de ovinos, quanto a produção de toxina, bem como a resistência aos antibióticos beta-lactâmicos. No presente estudo, dentre os quarenta isolados, foi observada a predominância do subtipo Stx1 (22/40), seguido do subtipo Stx1+ Stx2 (11/40), enquanto o grupo menos prevalente foi Stx2 (7/40). A resistência fenotípica aos antibióticos beta-lactâmicos foi observada em 50% (20/40) das cepas analisadas, formando dois grupos, um com isolados resistentes e outro de isolados não resistentes. A citotoxicidade dos isolados não variou entre os grupos. Alguns isolados multirresistentes, além de possuírem essa característica, produziram quantidades significativas de toxinas. Isto conclui, que os mecanismos de resistencia antimicrobiana, por meio de beta-lactamases estão presentes em STEC de ovinos, e que a citotoxicidade desses isolados é variável com relação a esta resistência.(AU)

Proteinograma de ovelhas imunizadas com veneno detoxificado do escorpião Tityus serrulatus / Proteinogram of immunized sheep with detoxified Tityus serrulatus scorpion venom

Background: Scorpionism is a worldwide medical issue, especially relevant in the tropical and subtropical countries. Tityusserrulatus is the species responsible for most cases in Brazil. Antivenom administration to victims is the sole specific therapyobtained from donor animals. Most of these donors suffer with symptoms of the poisoning, debilitating their health andreducing their life expectancy. The aim of the present research was to evaluate whether the immunogens prepared fromthe crude and detoxified venom of T. serrulatus promoted different changes in fractionated sheep plasma proteins, duringa scorpion antivenom serum production.Materials, Methods & Results: Twelve sheep, healthy, mean weight of 30 kg, were distributed into 3 groups (n = 4): G1(control), G2 (crude venom) and G3 (detoxified venom). The adopted immunization protocol (first cycle) had 6 doses, 3using Freund’s adjuvant, with a 21-day interval between each one (day 0, 22 and 43), and 3 doses with no adjuvant (booster)and 0.2 mg of antigen (reinforcement), spaced 3 days between each other (day 50, 53 and 56). Group control (G1) received6 immunizations with phosphate buffered saline (PBS) associated with Freund’s adjuvant (1:1), while the other 2 groupsreceived 0.5 mg of venom (G2) and detoxified venom (G3), respectively, diluted in PBS, associated with the Freund adjuvant. The boosters were 1/3 of the initial dose, diluted only PBS. At baseline (T0) and at 24 and 48 h after immunization,all animals underwent clinical examinations. Blood samples were collected at day 0, 22, 43, 53 and 56 for proteinogramanalysis. Total protein, albumin and globulins fractions were measured. Plasma albumin concentration at T0 ranged from3.41-4.86 g/dL, with a mean value of 4.12 g/dL. There was no statistical difference between...

First study of Brucella ovis antibodies in purebred sheep flocks in the State of Parana, Brazil / [Primeiro estudo de anticorpos para Brucella ovis em rebanhos de ovinos puros de Origem, no estado do Paraná, Brasil]

Brucella ovis, a non-zoonotic species, is the etiological agent of ovine brucellosis, an infectious disease of clinical or subclinical occurrence in sheep flocks. Until then, there is no serological study of anti-Brucella ovis antibodies in purebred sheep herds. This study aimed to determine the presence of anti-Brucella ovis antibodies in purebred sheep flocks with breeding purposes from Parana State. Blood samples from 728 animals, of which 563 were females and 165 males, between 8 and 56 months of age from the six major sheep producing mesoregions of Parana, were submitted to detection of anti-Brucella ovis antibodies by the Agar Gel Immunodiffusion technique using an antigen from the bacteria Brucella ovis (Reo 198). The results indicate the presence of this disease in purebred sheep from Parana State in a low occurrence of 0.27% (2/728). The only two positive animals were rams, Santa Inês breed, from the same flock in the East Center region of Parana, without clinical disease. In conclusion, Brucella ovis is present in purebred sheep in Parana State, Brazil, and this low occurrence may have occurred due to rigorous breeding systems that may contribute to reduce the transmission of this disease.(AU)

Brucella ovis, espécie não zoonótica, é o agente etiológico da brucelose ovina, doença infecciosa de ocorrência clínica ou subclínica. Atualmente, não existe estudo sorológico de anticorpos anti-Brucella ovis em rebanhos de ovinos puros de origem. Este estudo teve como objetivo determinar a presença de anticorpos anti-Brucella ovis em rebanhos ovinos de raça pura de origem, com fins reprodutivos do estado do Paraná. Amostras de sangue de 728 animais, sendo 563 fêmeas e 165 machos, entre oito e 56 meses de idade, pertencentes a seis principais mesorregiões produtoras de ovinos no Paraná, foram submetidas à detecção de anticorpos anti-Brucella ovis pela técnica de imunodifusão em ágar gel usando-se um antígeno da bactéria Brucella ovis (Reo 198). Os resultados indicam a presença da doença em ovinos puros de origem do estado do Paraná em baixa ocorrência de 0,27% (2/728). Os dois únicos animais positivos foram reprodutores da raça Santa Inês, do mesmo rebanho da região Centro Leste do Paraná, sem manifestação clínica. Em conclusão, Brucella ovis está presente em ovinos puros de origem no estado do Paraná, e essa baixa ocorrência pode ter ocorrido devido a sistemas rigorosos de criação, que podem contribuir para a redução da transmissão dessa doença.(AU)

Effect of supplementation with core homeopathic on the serum immunological profile of pregnant ewes and lambs / Efeito da suplementação com complexo homeopático no perfil imunológico sérico de ovelhas gestantes e cordeiros

Few information is available about the optimal period for colostrum absorption in the bowel of newborn lambs. The aim of the current study was to investigate the effects of using supplementation with commercial core homeopathic during pregnancy and early lactation on 18 pregnant Santa Inês ewes presenting body condition score ≥ 2.5. Pregnant animals were randomly divided in two groups; one was used as control (control group) and the other was supplemented on a daily basis with 30 g of homeopathic product with Natrum muriaticum 10-60, Calcium carbonicum 10-30, Silicea terra 10-400 and Hypothalamus 10-30 per animal, mixed to the concentrate during pregnancy and early lactation (treatment group). The Total Protein (TP) and globulin (GLB) values found in ewes and lambs belonging to the treatment group were higher than and different from those recorded for the control group. The serum albumin (ALB) concentration did not differ between groups of ewes but was different in lambs. TP and GLB evolution in lambs and ewes throughout the seven experimental days showed the animals in the treatment group trend to present higher TP and GLB values in all experimental periods; supplemented animals showed trend to progressive increase in both variables until day 3; whereas non-supplemented animals showed such trend until day 2 (when they reached their peak). The commercial core homeopathic to the animals during pregnancy and early lactation resulted in higher TP and GLB concentrations in the serum samples collected from ewes and lambs.

Poucas informações estão disponíveis sobre o período ideal para a absorção do colostro no intestino de neonatos ovinos. O objetivo do presente estudo é investigar os efeitos da suplementação com complexo homeopático comercial durante a gestação e início da lactação em 18 ovelhas Santa Inês gestantes com escore de condição corporal ≥ 2,5. Os animais gestantes foram divididos aleatoriamente em dois grupos; um controle (grupo controle) e outro suplementado diariamente com 30 g de produto homeopático com Natrum muriaticum 10-60, Calcium carbonicum 10-30, Silicea terra 10-400 and Hypothalamus 10-30 por animal, misturado ao concentrado durante a gestação e início da lactação (grupo tratamento). Os valores de Proteína Total (PT) e globulina (GLB) encontrados em ovelhas e cordeiros pertencentes ao grupo tratamento foram superiores e dife-rentes dos registrados para o grupo controle. A concentração de albumina sérica (ALB) não diferiu entre os grupos de ovelhas, mas foi diferente nos cordeiros. A evolução de PT e GLB em cordeiros e ovelhas ao longo dos 7 dias experimentais demons-trou que os animais do grupo tratamento tendem a apresentar maiores valores de PT e GLB em todo o período experimen-tal; os animais suplementados apresentaram tendência de aumento progressivo em ambas as variáveis até o dia 3; enquanto os animais não suplementados mostraram tal tendência até o dia 2 (quando atingiram o pico). O núcleo homeopático comercial fornecido para os animais durante a gestação e início da lactação resultou em maiores concentrações de PT e GLB nas amos-tras de soro coletadas de ovelhas e cordeiros.

Effect of supplementation with core homeopathic on the serum immunological profile of pregnant ewes and lambs / Efeito da suplementação com complexo homeopático no perfil imunológico sérico de ovelhas gestantes e cordeiros

Few information is available about the optimal period for colostrum absorption in the bowel of newborn lambs. The aim of the current study was to investigate the effects of using supplementation with commercial core homeopathic during pregnancy and early lactation on 18 pregnant Santa Inês ewes presenting body condition score ≥ 2.5. Pregnant animals were randomly divided in two groups; one was used as control (control group) and the other was supplemented on a daily basis with 30 g of homeopathic product with Natrum muriaticum 10-60, Calcium carbonicum 10-30, Silicea terra 10-400 and Hypothalamus 10-30 per animal, mixed to the concentrate during pregnancy and early lactation (treatment group). The Total Protein (TP) and globulin (GLB) values found in ewes and lambs belonging to the treatment group were higher than and different from those recorded for the control group. The serum albumin (ALB) concentration did not differ between groups of ewes but was different in lambs. TP and GLB evolution in lambs and ewes throughout the seven experimental days showed the animals in the treatment group trend to present higher TP and GLB values in all experimental periods; supplemented animals showed trend to progressive increase in both variables until day 3; whereas non-supplemented animals showed such trend until day 2 (when they reached their peak). The commercial core homeopathic to the animals during pregnancy and early lactation resulted in higher TP and GLB concentrations in the serum samples collected from ewes and lambs.(AU)

Poucas informações estão disponíveis sobre o período ideal para a absorção do colostro no intestino de neonatos ovinos. O objetivo do presente estudo é investigar os efeitos da suplementação com complexo homeopático comercial durante a gestação e início da lactação em 18 ovelhas Santa Inês gestantes com escore de condição corporal ≥ 2,5. Os animais gestantes foram divididos aleatoriamente em dois grupos; um controle (grupo controle) e outro suplementado diariamente com 30 g de produto homeopático com Natrum muriaticum 10-60, Calcium carbonicum 10-30, Silicea terra 10-400 and Hypothalamus 10-30 por animal, misturado ao concentrado durante a gestação e início da lactação (grupo tratamento). Os valores de Proteína Total (PT) e globulina (GLB) encontrados em ovelhas e cordeiros pertencentes ao grupo tratamento foram superiores e dife-rentes dos registrados para o grupo controle. A concentração de albumina sérica (ALB) não diferiu entre os grupos de ovelhas, mas foi diferente nos cordeiros. A evolução de PT e GLB em cordeiros e ovelhas ao longo dos 7 dias experimentais demons-trou que os animais do grupo tratamento tendem a apresentar maiores valores de PT e GLB em todo o período experimen-tal; os animais suplementados apresentaram tendência de aumento progressivo em ambas as variáveis até o dia 3; enquanto os animais não suplementados mostraram tal tendência até o dia 2 (quando atingiram o pico). O núcleo homeopático comercial fornecido para os animais durante a gestação e início da lactação resultou em maiores concentrações de PT e GLB nas amos-tras de soro coletadas de ovelhas e cordeiros.(AU)

Haemonchus contortus infection induces a variable immune response in resistant and susceptible Pelibuey sheep.

The immune response and phenotypic characteristics of Pelibuey lambs were analysed after the induction of a Haemonchus contortus trickle infection. Male lambs (n = 29; 20 kg live weight) were infected with 100 H. contortus infective larvae per kg of live weight on day 3, 5 and 7 of the experiment. The number of eggs per gram (epg), seven haematological parameters and the immunoglobulin A (IgA) level were analysed for 56 experimental days. In addition, histopathological samples from the fundic abomasal region and the relative expression of 10 immune-related genes from 15 infected and three non-infected lambs were analysed at day 0 and 49 of the experiment. The epg count and some haematological parameters (leucocytes, red blood cells, haemoglobin and total protein) with statistically significant differences (P < 0.01) were used to identify nine resistant and 20 susceptible lambs (1166 ±â€¯1071 and 3171 ±â€¯1463 epg, respectively). Moreover, acute infiltration of immune cells and parasitic granuloma formation were observed in susceptible lambs; the resistant group had moderate inflammatory cell infiltration. With respect to relative gene expression, resistant lambs showed upregulation (P < 0.001) of 10 genes, from 2.2 to 15.99 fold. Moreover, there was a strong indirect correlation (P < 0.05) between the epg count and interleukin 5 (IL5) gene expression. By contrast, there was an average 0.34 fold downregulation in nine of the immune-related genes (P ≤ 0.05) in susceptible lambs (the only exception was Fc fragment of IgE receptor Ia [FCER1A] upregulation). In addition, there was a direct correlation (P ≤ 0.05) between the epg count and the expression of IL8, which encodes an inflammatory chemokine. In conclusion, this study showed differential IL5 and IL8 gene expression during haemonchosis in resistant and susceptible Pelibuey lambs, respectively, together with a variable immune response based on histopathological and haematological parameters.

Bacillus toyonensis BCT-7112T transient supplementation improves vaccine efficacy in ewes vaccinated against Clostridium perfringens epsilon toxin.

AIM: The aim of the present study was to examine the vaccine immune response in ewes supplemented with Bacillus toyonensis BCT-7112T during a period of 5-day supplementation before vaccination against a recombinant Clostridium perfringens epsilon toxin (rETX). METHODS AND RESULTS: Ewes were vaccinated with 200 µg of rETX adjuvanted with 10% aluminium hydroxide. The treat group was orally supplemented with B. toyonensis BCT-7112T (3 × 108 viable spores) for 5 days prior to the first and second vaccination. Ewes supplemented with B. toyonensis BCT-7112T showed higher neutralizing antibody titres than the non-supplemented ewes (P < 0·05), with an increase in serum levels for total IgG anti-rETX by 3·2-fold (P < 0·0001), and for both IgG isotypes IgG1 and IgG2 by 2·1-fold and 2·3-fold (P < 0·01), respectively, compared with the control group. The peripheral blood mononuclear cells of ewes in the supplemented group had a higher (P < 0·05) cytokine mRNA transcription levels for IL-2 (6·4-fold increase), IFN-γ (2·9-fold increase) and transcription factor Bcl6 (2·3-fold increase) compared with the control group. CONCLUSION: We conclude that a 5 days of supplementation with B. toyonensis BCT-7112T prior vaccination is sufficient to significantly improve the humoral immune response of ewes against C. perfringens recombinant ETX vaccine. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings open a new perspective in the utilization of B. toyonensis BCT-7112T as an immunomodulator since a 5 days period of probiotic supplementation is sufficient to improve the vaccine immune response.

The effect of diets supplemented with Coriandrum sativumseeds on carcass performance, immune system, blood metabolites, rumen parameters and meat quality of lambs

A trial was conducted to investigate the effects of dietary Coriandrum sativum seeds on carcass performance, immune system, blood metabolites, rumen parameters and meat quality of Lambs. 16 Sanjabi lambs of 27 ± 5.1 kg during post-weaning (97 d of age) period were randomly selected. Four diets including 0, 1, 3, and 5% coriander seeds, replaced by Alfa alfa in the diet, were considered. A 30:70 alfalfa hay: concentrate diet for a period of time (97 to 187 d of age) was used. The results showed that feed intake was significantly increased by adding coriander seeds, linearly. There was no significant difference for apparent digestibility of crude protein, crude fat, neutral and acidic detergent fiber, crude ash, rumen fluid pH and ammonia nitrogen at 0, 2 and 4 h after feeding, Meat dry matter, ash, crude protein and fat, and the meat’s fatty acid profile (p > 0.05). Dietary coriander seeds had a significant effect on neutrophils, lymphocytes, monocytes and eosinophil’s (in days 7 and 14 of trial) and blood metabolites at the middle of trial. Obtained results suggested that supplementation of coriander seed may have limited effects on nutrient digestibility, ruminal parameters, meat quality, blood cells and metabolites.

Anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Paraná state, South Brazil: prevalence and associated factors / Anticorpos anti-Toxoplasma gondii e anti-Neospora caninum em ovinos do estado do Paraná, sul do Brasil: prevalência e fatores associados

The aim of this study was to evaluate the seroprevalence and factors associated with the presence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Paraná state. The detection of antibodies for T. gondii and N. caninum was performed by homemade and commercial indirect ELISA, respectively. Multiple logistic regression analysis was used to verify the factors associated with the seroprevalence. Antibodies anti-T. gondii and anti-N. caninum were observed in 42.7% and in 17.6% of the animals, respectively. The protective factors associated to seropositive were some level of confinement (full or semi-extensive confinement) (OR=0.53) for T. gondii and use of skilled labor (OR=0.64) for N. caninum. The risk factors were presence of cats (OR=1.75) for T. gondii and feeding of dogs with sheep placental remains (OR=1.79) for N. caninum. In addition, to presenting a significant and simultaneous seroprevalence for both agents (9.9% of the animals), the results also indicate that deficiencies in management and environmental sanitation, the presence of reservoirs, and types of exploitation enhance the seropositivity. Thus, studies like this might support sanitary programs and public policies for the prevention of T. gondii and N. caninum in the sheep herds of Paraná state.(AU)

O objetivo deste estudo foi avaliar a soroprevalência e os fatores de risco associados à presença de anticorpos contra Toxoplasma gondii e Neospora caninum em ovinos do Paraná. A detecção dos anticorpos para T. gondii e N. caninum foi realizada, usando-se testes de ELISA indireto caseiro e comercial, respectivamente. A análise de regressão logística múltipla foi usada para verificar os fatores de risco associados à soroprevalência. Anticorpos anti-T. gondii e anti-N. caninum foram observados em 42,7% e em 17,6% dos animais, respectivamente. Os fatores de proteção associados à soropositividade foram algum nível de confinamento (confinamento ou semiextensivo) (OR=0,53) para T. gondii e presença de mão de obra especializada para N. caminum (OR=0,64). Os fatores de risco foram presença de gatos para T. gondii (OR=1,75) e alimentação de cães com resíduos placentários (OR=1,79) para N. caninum. Além de apresentar uma significante e simultânea soroprevalência para ambos os agentes (9,9% dos animais), os resultados também indicam que as deficiências em gestão e saneamento ambiental, a presença de reservatórios e os tipos de exploração favorecem a soropositividade. Dessa forma, estudos como estes podem auxiliar em programas sanitários e políticas públicas para a prevenção de T. gondii e N. caninum nos rebanhos ovinos do Paraná.(AU)

The effect of diets supplemented with Coriandrum sativumseeds on carcass performance, immune system, blood metabolites, rumen parameters and meat quality of lambs

A trial was conducted to investigate the effects of dietary Coriandrum sativum seeds on carcass performance, immune system, blood metabolites, rumen parameters and meat quality of Lambs. 16 Sanjabi lambs of 27 ± 5.1 kg during post-weaning (97 d of age) period were randomly selected. Four diets including 0, 1, 3, and 5% coriander seeds, replaced by Alfa alfa in the diet, were considered. A 30:70 alfalfa hay: concentrate diet for a period of time (97 to 187 d of age) was used. The results showed that feed intake was significantly increased by adding coriander seeds, linearly. There was no significant difference for apparent digestibility of crude protein, crude fat, neutral and acidic detergent fiber, crude ash, rumen fluid pH and ammonia nitrogen at 0, 2 and 4 h after feeding, Meat dry matter, ash, crude protein and fat, and the meats fatty acid profile (p > 0.05). Dietary coriander seeds had a significant effect on neutrophils, lymphocytes, monocytes and eosinophils (in days 7 and 14 of trial) and blood metabolites at the middle of trial. Obtained results suggested that supplementation of coriander seed may have limited effects on nutrient digestibility, ruminal parameters, meat quality, blood cells and metabolites.(AU)

Immunogenicity in sheep of Uruguayan commercial vaccines against bovine alphaherpesvirus 1, 5 and bovine pestiviruses / Imunogenicidade em ovelhas de vacinas comerciais Uruguaias para alfaherpesvírus bovino 1, 5 e pestivírus de bovinos

The serological responses induced by four commercial inactivated Uruguayan vaccines against bovine alphaherpesviruses (BoHV)-1 and -5 and bovine pestiviruses (BVDV-1, BVDV-2, and HoBiPeV) were evaluated in sheep. Thirty-seven sheep were immunized twice (day 0 and 25) and their serum samples were tested at different intervals (days 0, 25, 40, 60, and 90) post-vaccination (PV). Among the four vaccines tested, only one (G4) could induce the production of moderate neutralizing antibody titers against BoHV-1 and -5 and BVDV-1 and -2. The G3 vaccine showed a neutralizing serological response against the bovine alphaherpesviruses only. The G1 and G2 vaccines produced extremely low levels of antibodies in a few vaccinated animals only (geometric mean titers (GMT) 2.2). Similar levels of immunological responses were induced by the G4 vaccine against BoHV-1 and -5, and titers of neutralizing antibodies induced in approximately 70% of the animals are known to confer protection (GMT > 8). For bovine pestiviruses, the vaccine stimulated response of G4 against BVDV-2 was higher compared to that against BVDV-1, and extremely low for HoBiPeV. The peak of neutralizing antibodies to BoHV-1 and BVDV-1 was observed on days 40 and 60 PV, respectively. Thereafter, a remarkably decrease in neutralizing antibody response was observed at day 90 PV. These results demonstrated that tested commercial Uruguayan vaccines did not induce a serological response of adequate magnitude and duration. Thus, it is important to periodically review formulations and compositions of commercial vaccines against bovine alphaherpesviruses and pestiviruses.(AU)

A resposta sorológica induzida por quatro vacinas comerciais uruguaias inativadas contra os alfaherpesvírus bovinos (BoHV-1 e -5) e pestivírus de bovinos (BVDV-1, BVDV-2 e HoBiPeV) foi avaliada em ovinos. Os animais foram imunizados duas vezes (dia 0 e dia 25) e o soro testado em diferentes intervalos (dias 0, 25, 40, 60 e 90) após a vacinação (PV). Dentre as quatro vacinas testadas, apenas uma (G4) apresentou títulos de anticorpos neutralizantes moderados para os BoHV-1 e -5, BVDV-1 e 2. A vacina G3 apresentou resposta somente para alfaherpesvírus bovinos. As vacinas G1 e G2 estimularam resposta somente em alguns animais vacinados. Para a vacina G4, observou-se que a resposta imunológica frente ao BoHV-1 e 5 foi semelhante e pelo menos 70% dos animais apresentaram níveis protetivos de anticorpos neutralizantes. Para os pestivírus bovinos, a vacina G4 estimulou resposta para o BVDV-2 mais elevada quando comparada com o BVDV-1, e quase que indetectável para HoBiPeV. O pico de anticorpos neutralizantes para o BoHV-1 foi observado no dia 40 PV e no dia 60 PV para o BVDV-1. Após isso, observou-se um decréscimo considerável na resposta de anticorpos neutralizantes. Os resultados demonstraram que vacinas comerciais uruguaias testadas não induziram resposta sorológica de magnitude e duração adequadas. Assim, ressalva-se a importância de rever periodicamente a formulação e composição das vacinas comerciais para alfaherpesvírus e pestivírus bovinos.(AU)

Saccharomyces cerevisiae (YT001) supplementation for the control of Haemonchus contortus and modulation of the immune response of sheep.

Studies aiming at the development and evaluation of alternative methods to minimise losses caused by the gastrointestinal nematode Haemonchus contortus are extremely important. Such research is essential, given the high morbidity rates among sheep and the significant mortality rates of lambs, allied to the low efficacy of commercial products for the control of this parasite. The purpose of this study was to evaluate the effect of the Saccharomyces cerevisiae (YT001 - YEASTECH) on the control of H. contortus and its modulation of the immune response in experimentally infected sheep. Eighteen sheep were divided into two groups. Group 1, the control group, comprised animals infected with H. contortus and supplemented with distilled water, while Group 2, the treated group, consisted of animals infected and supplemented with S. cerevisiae (400 million cfu/day of suspension for 49 days). The following parasitological parameters were evaluated: number of eggs per gram of faeces, number of infective larvae (L3) recovered per faecal culture, and parasitic load of the abomasum. The following immunological parameters were quantified: immunoglobulin (Ig)A in the mucous secretions and serum IgG; cytokines interleukin (IL)-4, IL-5 and IL-10; number of eosinophils in the abomasal mucosa and groups of cells positive for the markers: MHCII, CD4+CD25+, CD5+CD8+, WC4, CD5+CD4+, CD8+CD11b+ and CD5+WC1 by whole blood flow cytometry. The results revealed a significant decrease (P<0.05) in the number of larvae and significantly higher serum IgG levels (P<0.05) in the group supplemented with S. cerevisiae. The supplemented animals showed significantly larger numbers of eosinophils (P<0.05), as well as more cells positive for MHCII, CD4+CD25+, CD5+CD8+ than the control animals. This study confirmed the beneficial action of S. cerevisiae on the host immune response to H. contortus, as evidenced mainly by the smaller number of L3 recovered from the faeces of sheep supplemented with S. cerevisiae.

Infección experimental en ovinos con el virus de leucosis bovina: dosis mínima de células BLV-FLK y virus libre de células, y actividad neutralizante de anticuerpos naturales / Experimental infection of sheep with Bovine leukemia virus (BLV): Minimum dose of BLV-FLK cells and cell-free BLV and neutralization activity of natural antibodies

Abstract Bovine leukemia virus (BLV) is an important cattle pathogen that causes major economic losses worldwide, especially in dairy farms. The use of animal models provides valuable insight into the pathogenesis of viral infections. Experimental infections of sheep have been conducted using blood from BLV-infected cattle, infectious BLV molecular clones or tumor-derived cells. The Fetal Lamb Kidney cell line, persistently infected with BLV (FLK-BLV), is one of the most commonly used long-term culture available for the permanent production of virus. FLK-BLV cells or the viral particles obtained from the cell-free culture supernatant could be used as a source of provirus or virus to experimentally infect sheep. In this report, we aimed to determine the minimum amount of FLK-BLV cells or cell-free supernatant containing BLV needed to produce infection in sheep. We also evaluated the amount of antibodies obtained from a naturally-infected cow required to neutralize this infection. We observed that both sheep experimentally inoculated with 5000 FLK-BLV cells became infected, as well as one of the sheep receiving 500 FLK-BLV cells. None of the animals inoculated with 50 FLK-BLV cells showed evidence of infection. The cell-free FLK-BLV supernatant proved to be infective in sheep up to a 1:1000 dilution. Specific BLV antibodies showed neutralizing activity as none of the sheep became infected. Conversely, the animals receiving a BLV-negative serum showed signs of BLV infection. These results contribute to the optimization of a sheep bioassay which could be useful to further characterize BLV infection.

Resumen El virus de la leucosis bovina (bovine leukemia virus [BLV]) es un importante agente patógeno del ganado que causa importantes pérdidas económicas en todo el mundo, especialmente en los rodeos lecheros. El uso de modelos animales proporciona información valiosa sobre la patogénesis de las infecciones virales. Se realizaron infecciones experimentales en ovejas usando sangre de bovinos infectados con BLV, clones moleculares de BLV infecciosos o células derivadas de tumores. La línea celular Fetal Lamb Kidney, persistentemente infectada con el BLV (FLK-BLV), es uno de los cultivos a largo plazo más utilizados para la producción permanente de virus. Las células FLK-BLV o las partículas virales obtenidas del sobrenadante del cultivo libre de células podrían usarse como fuente de provirus o de virus para infectar experimentalmente ovejas. En este trabajo, nuestro objetivo fue determinar la cantidad mínima de células FLK-BLV o de sobrenadante libre de células que contiene BLV necesaria para producir infección en ovejas. También evaluamos la cantidad de anticuerpos bovinos anti-BLV necesaria para neutralizar la infección. Observamos que las dos ovejas inoculadas experimentalmente con 5000 células FLK-BLV se infectaron, y que una de las dos ovejas que recibieron 500 células FLK-BLV se infectó. Ninguno de los animales inoculados con 50 células FLK-BLV mostró evidencia de infección. El sobrenadante FLK-BLV libre de células demostró ser infectivo en ovejas hasta la dilución 1:1000. Los anticuerpos BLV específicos mostraron actividad neutralizante, ya que ninguna de las ovejas se infectó. Por el contrario, los animales que recibieron un suero BLV negativo mostraron signos de infección por BLV. Estos resultados contribuyen a la optimización de un bioensayo en ovejas útil para caracterizar la infección por BLV.

Detection of Echinococcus granulosus sensu lato infection by using extracts derived from a protoscoleces G1 cell line.

Cystic echinococcosis (CE) can be diagnosed by means of several serological approaches, but their results vary among laboratories due to the molecular characteristics of the reference antigens used. Thus, this study aimed to address both the relevance of an EGPE cell line previously obtained from Echinococcus granulosus protoscoleces G1 and the complexity of the immune response by using two different in vitro growth stages as separate sources of parasite antigens. The serum reactivity was investigated by western blotting (WB) in 21 CE patients from an endemic area in a matched case-control design and also in seven experimentally infected sheep and five healthy control sheep. EGPE-antigen-human serum sensitivity by WB was higher than that of hydatid fluid (HF) WB, ELISA and DD5 (P < .05, Chi-square test). EGPE protein extract was immunogenic in mice and hyperimmune plasma reacted with HF proteins, and AgB2 expression was detected by molecular analysis. Proteins of 37 to 60 kDa were recognized by 95.24% of the CE patients' sera but, with poor specificity. Statistically significant differences were found between serum protein extract recognition at 7 and 20 days of cell growth. The EGPE cell line is a laboratory source of antigens for improvement of CE serological diagnosis.

Determinação da frequência de anticorpos anti - T. gondii em caprinos e ovinos abatidos para o consumo humano na região metropolitana de Recife-PE / Determination of frequency of anti - T. gondii antibodies in goats and sheep abolished for human consumption in the metropolitan region of Recife – PE

Ovinos e caprinos são suscetíveis a infecções por Toxoplasma gondii e importantes na transmissão a humanos através do consumo de carnes crua ou mal passada. Objetivou-se pesquisar o T. gondii nos animais abatidos em abatedouro situado na região metropolitana do Recife-PE, identificando anticorpos anti - T. gondii, sob a metodologia da Imunofluorescência Indireta (RIFI). Analisou-se 93 amostras, 66 amostras de soro ovino e 27 amostras de soro caprino. Nas ovinas, obteve-se 60,6% (40/66) positivas, sendo 31,81% (21/66) para titulação de 1:64; 10,6% (7/66) para 1:128; 7,57% (5/66) para 1:256; 6,06% (4/66) para 1:512; 4,54% (3/66) para 1:1024. Nas caprinas, obteve-se 62,96% (17/27) positivas, sendo 37,03% (10/27) para titulação de 1:64; 14,81% (4/27) para 1:128; 7,4% (2/27) para 1:256 e 3,7% (1/27) para 1:512, sem positividade para 1:1024. Conclui-se que há uma frequência expressiva de IgG anti - T. gondii nos soros provenientes deste abatedouro, tornando-se questão de saúde pública.

Determinação da frequência de anticorpos anti - T. gondii em caprinos e ovinos abatidos para o consumo humano na região metropolitana de Recife-PE / Determination of frequency of anti - T. gondii antibodies in goats and sheep abolished for human consumption in the metropolitan region of Recife PE

Ovinos e caprinos são suscetíveis a infecções por Toxoplasma gondii e importantes na transmissão a humanos através do consumo de carnes crua ou mal passada. Objetivou-se pesquisar o T. gondii nos animais abatidos em abatedouro situado na região metropolitana do Recife-PE, identificando anticorpos anti - T. gondii, sob a metodologia da Imunofluorescência Indireta (RIFI). Analisou-se 93 amostras, 66 amostras de soro ovino e 27 amostras de soro caprino. Nas ovinas, obteve-se 60,6% (40/66) positivas, sendo 31,81% (21/66) para titulação de 1:64; 10,6% (7/66) para 1:128; 7,57% (5/66) para 1:256; 6,06% (4/66) para 1:512; 4,54% (3/66) para 1:1024. Nas caprinas, obteve-se 62,96% (17/27) positivas, sendo 37,03% (10/27) para titulação de 1:64; 14,81% (4/27) para 1:128; 7,4% (2/27) para 1:256 e 3,7% (1/27) para 1:512, sem positividade para 1:1024. Conclui-se que há uma frequência expressiva de IgG anti - T. gondii nos soros provenientes deste abatedouro, tornando-se questão de saúde pública.(AU)

Brazilian red pepper fruit essential oil (Schinus terebinthifolius) may replace monensin in high concentrate diets for feedlot lambs / Óleo essencial dos frutos de aroeira (Schinus terebinthifolius) pode substituir a monensina em dietas com elevado teor de concentrado para cordeiros confinados

Essential oil (EO) from Brazilian red pepper fruit contains compounds with antimicrobial activity, and could be possible substitutes for the antibiotics commonly used in ruminant nutrition. The objectives of the present study were to evaluate the effects of the Brazilian red pepper fruit EO (Schinus terebinthifolius) as a substitute for monensin on performance, carcass characteristics and meat of lambs fed high concentrate diets. Forty-eight lambs were used, 24 males (20 ½ Dorper × ½ Santa Inês and 4 Santa Inês) and 24 females (24 ½ Dorper × ½ Santa Inês), with 21.54 ± 0.88 kg of initial body weight (BW) and 78 ± 2.4 days of age, in a randomized complete block design. The experiment lasted 56 days, divided into 2 periods of 28 days each. The treatments were defined by the inclusion in diets of 8 ppm of monensin (MON), and the doses 0.14% (14EO), 0.28% (28EO) and 0.42% (42EO) of red pepper fruit EO. The additives were included in a base diet with a 10:90 of forage to concentrate ratio. At the end of 56 days, 32 animals were slaughtered for the measurement of carcass parameters and meat composition. There was no interaction among treatments and periods for average daily gain (P = 0.08), DM intake (P = 0.36), feed efficiency (P = 0.24) and oocyst of Eimeria ssp. in feces (P = 0.46). The treatments did not affect (P > 0.05) the average daily gain (ADG), dry matter intake (DMI) and feed efficiency. Lambs fed diets containing monensin had less (P < 0.01) oocyst/g compared with the diet 14EO. There was no effect of diets on carcass characteristics. The treatments with higher doses of the Brazilian red pepper fruit EO had reduced mineral content of meat compared to monensin. The red pepper fruit EO demonstrated the potential to replace monensin in feedlot lambs fed high concentrate diets, maintaining performance and carcass characteristics. However, the monensin has greater capacity to control coccidiosis in feedlot lambs.

Os óleos essenciais (OE) dos frutos de aroeira possuem compostos com atividade antimicrobiana, sendo possíveis substitutos aos antibióticos comumente utilizados na nutrição de ruminantes. Os objetivos do presente estudo foram avaliar os efeitos da inclusão do óleo essencial de aroeira fruta (Schinus terebinthifolius) como substituto da monensina sobre o desempenho, características de carcaça e da carne de cordeiros alimentados com dietas contendo elevado teor de concentrado. Foram utilizados 48 cordeiros, 24 machos (20 ½ Dorper × ½ Santa Inês e 4 Santa Inês) e 24 fêmeas (24 ½ Dorper × ½ Santa Inês), com peso inicial de 21,54 ± 0,88 kg e 78 ± 2,4 dias de idade, em delineamento de blocos completos casualizados. O experimento teve duração de 56 dias, divididos em 2 períodos de 28 dias cada. Os tratamentos foram definidos pela inclusão na dieta de 8 ppm de monensina sódica (MON) e as doses de 0,14% (14EO), 0,28% (28EO) e 0,42% (42EO) de óleo essencial dos frutos da aroeira. As dietas experimentais foram compostas por 10% de volumoso e 90% de concentrado. Ao final dos 56 dias, 32 animais foram abatidos para a mensuração dos parâmetros de carcaça e análise química da carne. Não houve interação entre tratamento e período para o ganho médio diário (P = 0,08), consumo de MS (P = 0,36), eficiência alimentar (P = 0,24) e contagem de oocistos de Eimeria ssp. (P = 0,46). Não houve efeito (P > 0,05) dos tratamentos no ganho de peso médio diário (GMD), consumo de matéria seca (CMS) e eficiência alimentar (EA). Cordeiros alimentados com dietas contendo monensina tiveram menor (P < 0,01) contagem de oocistos/g de fezes comparado com a dieta 14OE. Não houve efeito das dietas sobre as características de carcaça. A inclusão de 0,28 e 0,42% de OE de aroeira fruto reduziram a concentração de matéria mineral da carne dos cordeiros comparados ao tratamento MON. O OE dos frutos da aroeira demonstrou capacidade de substituir a monensina, apresentando resultados similares com relação ao desempenho e características de carcaça. Entretanto, a monensina apresentou maior capacidade no controle de coccidiose

First molecular detection of Mycoplasma ovis (Hemotropic mycoplasmas) from Sheep in Brazil / Primeira detecção molecular de Mycoplasma ovis (Micoplasmas Hemotrópicos) em Ovinos no Brasil

Mycoplasma ovis is an emerging zoonotic pathogen with a worldwide distribution and can cause mild to severe hemolytic anemia, icterus, and poor weight gain in animals. Although M. ovis has been described in small ruminants worldwide, data on M. ovis in sheep in Brazil is unknown. The objective of the present study was to present the first report of hemotropic mycoplasma (HM) in sheep from Brazil. We evaluated factors associated with this infection, such age group, tick presence, and anemia. Blood samples were collected from 33 sheep from a farm in southern Brazil and screened for hemoplasmas using PCR. Out of 33 samples, 26 (78.8%) tested positive for M. ovis. The sequencing of positive samples showed 100% identity with multiple M. ovis 16S rDNA sequences. No association was observed between the presence of M. ovis and the FAMACHA© score (p = 0.620). Rhipicephalus (Boophilus) microplus (15/33, 45.4%) was the tick species found on the animals. No significant association between M. ovis infection and presence of ticks (p = 0.4134) and age group (p = 0.4221) was observed. This is the first report of M. ovis infection in sheep from Brazil and only the second report of this pathogen in sheep in Latin America.(AU)

Mycoplasma ovis é um patógeno zoonótico emergente com distribuição mundial e pode causar anemia hemolítica de leve a grave, icterícia e baixo ganho de peso em animais. Embora M. ovis tenha sido descrito em pequenos ruminantes em todo o mundo, os dados sobre M. ovis em ovinos no Brasil são desconhecidos. O objetivo deste estudo foi apresentar o primeiro relato de micoplasmas hemotrópicos em ovinos no Brasil. Avaliamos os fatores associados a essa infecção, como faixa etária, presença de carrapatos e anemia. Amostras de sangue foram coletadas de 33 ovelhas de uma fazenda no sul do Brasil e testadas para hemoplasmas usando a PCR. Das 33 amostras, 26 (78,8%) apresentaram resultado positivo. O sequenciamento das amostras positivas mostrou 100% de identidade com múltiplas sequências de M. ovis 16S rDNA. Não foi observada associação entre a presença de M. ovis e o escore FAMACHA© (p = 0,620). Rhipicephalus (Boophilus) microplus (15/33, 45,4%) foi a espécie de carrapato encontrada nos animais. Não houve associação significativa entre infecção por M. ovis e presença de carrapatos (p = 0,4134) e faixa etária (p = 0,4221). Este é o primeiro relato de infecção por M. ovis em ovinos no Brasil e o segundo relato deste patógeno em ovinos na América Latina.(AU)

Temporal evolution of anti-Clostridium antibody responses in sheep after vaccination with polyvalent clostridial vaccines.

Polyvalent clostridial vaccines, composed of a complex mixture of toxoids from up to 9 different species, are highly effective in controlling clostridial diseases in cattle and sheep. Commercially available vaccines usually state that in normal field conditions two doses administered 4 to 6 weeks apart elicit protective antibody levels that will last for one year. However, studies on the development and duration of the antibody response against the different Clostridium species in target animals are scarce and only partial. Evaluating the temporal evolution of the antibody responses upon vaccination in target species is relevant to understand the bases of protective immunity induced by these vaccines and to develop new optimized vaccines. Here, we assessed the antibody response in sheep against each Clostridium component of two different 9-valent Clostridial vaccines over the period of one year. One vaccine was a commercially available vaccine and the other was an experimental vaccine prepared by us with the same antigens that we used to set up a specific ELISA for each Clostridium species. Both vaccines showed similar results, irrespectively of the origin of the antigens used for the ELISAs, with antibody titers that peaked at day 36 after vaccination and large inter individual variations in the magnitude of the response. Antibody titers were maintained up to 90 days and then markedly decreased, becoming even undetectable in some animals 6 months after vaccination. Given that the current scheme of yearly revaccination has largely shown to be effective at controlling the burden of disease, our results strongly suggest that circulating antibody levels cannot completely explain the protective immunity elicited by these vaccines, and prompt for further studies into the correlates of protection of clostridial vaccines.