Molecular confirmation of ovine herpesvirus 2-induced malignant catarrhal fever lesions in cattle from Rio Grande do Norte, Brazil

Molecular findings that confirmed the participation of ovine herpesvirus 2 (OVH-2) in the lesions that were consistent with those observed in malignant catarrhal fever of cattle are described. Three mixed-breed cattle from Rio Grande do Norte state demonstrated clinical manifestations that included mucopurulent nasal discharge, corneal opacity and motor incoordination. Routine necropsy examination demonstrated ulcerations and hemorrhage of the oral cavity, corneal opacity, and lymph node enlargement. Significant histopathological findings included widespread necrotizing vasculitis, non-suppurative meningoencephalitis, lymphocytic interstitial nephritis and hepatitis, and thrombosis. PCR assay performed on DNA extracted from kidney and mesenteric lymph node of one animal amplified a product of 423 base pairs corresponding to a target sequence within the ovine herpesvirus 2 (OVH-2) tegument protein gene. Direct sequencing of the PCR products, from extracted DNA of the kidney and mesenteric lymph node of one cow, amplified the partial nucleotide sequences (423 base pairs) of OVH-2 tegument protein gene. Blast analysis confirmed that these sequences have 98-100% identity with similar OVH-2 sequences deposited in GenBank. Phylogenetic analyses, based on the deduced amino acid sequences, demonstrated that the strain of OVH-2 circulating in ruminants from the Brazilian states of Rio Grande do Norte and Minas Gerais are similar to that identified in other geographical locations. These findings confirmed the active participation of OVH-2 in the classical manifestations of sheep associated malignant catarrhal fever.

A clinical and hemogasometric survey of neonatal lambs

In order to provide adequate medical assistance to neonates, the extent of vitality impairment has to be investigated through complementary exams, as well as clinical assessment. This investigation aimed to identify the physiological changes that occur during neonatal adaptation and to develop a clinical approach that can be performed during the first hour of life in neonatal lambs born through vaginal eutocic labor. The neonatal vitality of 14 Santa Ines lambs was verified using the Apgar system and rectal temperature at birth and after 5 and 60 min after birth. From the total number of neonates, 7 lambs were randomly selected for blood gas analysis and glucose immediately at birth and 1 h after birth. The lambs had hypoglycemia immediately after birth, as well as acidosis due to metabolic and respiratory causes. Given their hypoxemia at birth, lambs immediately exhibit tachycardia and tachypnea. However, neonatal lambs reached Apgar score superior than 7 after 5 min of birth. Ovine neonates are relatively mature at birth, with adequate thermoregulation and active mechanisms to compensate for physiological acid-base imbalances. In conclusion, a systematic clinical examination of newborn sheep should include the implementation of the Apgar score coupled with the confirmation of any acid-base imbalances. Further research should evaluate neonatal adaptation to this critical period over a longer period of time. (C) 2012 Elsevier B.V. All rights reserved.

Post-thaw viability of in vivo-produced canine blastocysts cryopreserved by slow freezing

The objectives were to evaluate the reexpansion blastocoele rate, post-thaw viability, and in vitro development of canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol (GLY) or 1.5 m ethylene glycol (EG). Fifty-one in vivo-produced canine blastocysts were randomly allocated in two groups: GLY (n = 26) and EG (n = 25). After thawing, embryos from MO were immediately stained with the fluorescent probes propidium iodide and Hoechst 33 342 to evaluate cellular viability. Frozen-thawed embryos from M3 and M6 were cultured in SOFaa medium + 10% FCS at 38.5 degrees C under an atmosphere of 5% CO2 with maximum humidity, for 3 and 6 days, respectively, and similarly stained. The blastocoele reexpansion rate (24 h after in vitro culture) did not differ between GLY (76.5%) and EG (68.8%). Post-thaw viable cells rate were not significantly different between GLY and EG (66.5 +/- 4.8 and 57.3 +/- 4.8, respectively, mean +/- SEM), or among MO (62.3 +/- 5.7%), M3 (56.9 +/- 6.0%), and M6 (66.5 +/- 6.0%). In conclusion, canine blastocysts cryopreserved by slow freezing in 1.0 m glycerol or 1.5 m ethylene glycol, had satisfactory blastocoele reexpansion rates, similar post-thawing viability, and remained viable for up to 6 days of in vitro culture. (C) 2012 Elsevier Inc. All rights reserved.

Intestinal IgG uptake by small intestine of goat kid fed goat or lyophilized bovine colostrum

The immunoglobulin G (IgG) uptake and enterocyte nucleus position in the villous were studied in newborn goat kids fed goat or lyophilized bovine colostrum. Two groups of 15 newborn goat kids, each received 5% of body weight of goat colostrum (GC) or lyophilized bovine colostrum (LBC) containing 55 mg/mL of immunoglobulin G (IgG) at 0, 7 and 14 h of life. Three animals were sampled just after birth, receiving no colostrum intake, to be used as control. Samples of duodenum, medium jejunum and ileum were collected at 0, 18, 36 and 96 h of life. IgG vacuoles were not observed in the duodenum throughout the experiment regardless of all the experimental time points. In this segment, at 0, 18 and 36 h of life, nuclei were found in the apical, medial and basal positions in the enterocytes, and localized in the upper, medial and lower parts in the villous, respectively. At 96 h, a basal nuclei position was observed in the enterocytes, throughout the villous. In jejunum, IgG vacuoles were distributed along the villous at 18 and 36 h. In this segment at Oh the nuclei were positioned predominantly apically in the enterocytes, throughout the villous. At 18 and 36 h, no consistent nuclei pattern was verified: however at 96 h, the nuclei were positioned basally in the enterocytes, throughout the jejunal villous. In the ileum at 0, 18 and 36 h, a great number of vacuoles without IgG were verified in the medial-apical part of the villous. In this segment, at Oh of life and 96 h of life, the predominance of basal nuclei was observed. Nuclei were positioned in medial-apically part of the ileal enterocytes in the upper part of the villous at 18 and 36 h. It was found that the jejunal epithelium was the most important segment related to absorption process. The IgG absorption and nucleus position in the newborn goats were dependent on the small intestine segments and experimental time points, regardless of the colostrum source. GC or LCB. Considering the IgG uptake mechanism observed in the present study, the lyophilized bovine colostrum might be used instead of goat colostrum. (C) 2011 Elsevier B.V. All rights reserved.

Lyophilized bovine colostrum as a source of immunoglobulins and insulin-like growth factor for newborn goat kids

The objective of this study was to evaluate lyophilized bovine colostrum as an alternative source of passive immunity and insulin like growth factor I (IGF-I) for goat kids, considering newborns consuming non-maternal colostrum. Twenty-nine male newborns received 5% of body weight of lyophilized bovine (LBC) or goat colostrum (GC), both with 55 mg/mL of IgG, at 0, 7 and 14 h of life. Blood samples were collected at 0, 7, 14, 18, 24, 36, 48, 72 and 96 h of life to determine serum IgG, total protein (TP), IGF-I and apparent efficiency of IgG absorption at 7, 14, 18 and 24 h (AEA(7hr), AEA(14hr), AEA(18hr), AEA(total), respectively). In LBC, the values of serum IgG at 14, 18,24 and 48 h (13.1, 13.4, 14.1 and 14.6 mg/mL, respectively) were higher than the values at 0 and 7 h (0.04 and 6.9 mg/mL, respectively). In GC, the serum IgG at 18 h (9.3 mg/mL) was higher than the value at 7 h (5.5 mg/mL). AEA(7hr) and AEA(14hr) in LBC were the same (19.2 and 18.5%, respectively, P>0.05) and the values of AEA(18hr) and AEA(total), 9.3 and 9.5%, respectively, were equal and smaller than AEA(7hr), and AEA(14hr). In GC, AEA(7hr), 20.8%, was higher than AEA(14hr), 16.1% (P<0.05) and AEA(18hr), and AEA(total), 9.2 and 8.0%, respectively, were equal and smaller than AEA(7hr) and AEA(14hr). The serum TP and IGF-I were not affected by colostrum feeding. Considering the variables study in the present work, lyophilized bovine colostrum constitutes a promising alternative substitute to goat colostrum in newborn goat kids, since the supply of immunoglobulins and IGF-I was suitable for the kids. (c) 2012 Elsevier B.V. All rights reserved.

Goat kids' intestinal absorptive mucosa in period of passive immunity acquisition

Colostrum intake in newborn goat kids is essential for the acquisition of immunoglobulins (Ig) and influencing development of gastrointestinal mucosa. The present study investigated small intestine structure in the postnatal goat kid fed lyophilized bovine colostrum, an alternative source of antibodies to small ruminants, or goat colostrum using scanning electron microscopy technique. At 0,7 and 14 h of life 15 male newborns received 5% of body weight of lyophilized bovine colostrum (LBC) and 14 goat colostrum (GC), both with 55 mg/mL of IgG. Samples of duodenum, medium jejunum and ileum were collected at 18, 36 and 96 h of life. Three animals were sampled at birth without colostrum intake (0 h). The enteric tissues were analyzed for villi density (villi/cm(2)) and morphological characteristics. The villi density did not differ between treatment, sampling time and intestinal segments (P>0.05). The morphological characteristics were not different between LBC and GC in all segments. Duodenal villi were fingerlike, thick and short, and with different heights. Duodenal folds could also be verified. Frequent anastomoses in all sampling times were observed in this segment. In the jejunum, fingerlike villi, thin and thick, of different heights were observed in all sampling times as well as leaf-shaped villi. Vacuoles with colostrum were observed in the jejunum of goats sampled at 18 h of life. In ileum, fingerlike villi were observed in all sampling times. At 0 and 96 h of life, thick and low villi were verified while at 18 and 36 h the villi showed different heights and widths. At all sampling times, regularly cell extrusion processes were observed with grouped cells at the apex of the ileum villi and with isolated cells along the villi. In the first 4 days of goat kids' life the small intestine structure was unaffected by different sources of colostrum, goat or lyophilized bovine, and by the replacement of fetal enterocytes, which are able to absorb macromolecules, by adult-type ones. (C) 2011 Elsevier B.V. All rights reserved.

Maternal high-sodium intake alters the responsiveness of the renin-angiotensin system in adult offspring

Aims: The goal of the current study was to evaluate the impact of maternal sodium intake during gestation on the systemic and renal renin-angiotensin-aldosterone-system (RAAS) of the adult offspring. Main methods: Female Wistar rats were fed high- (HSD-8.0% NaCl) or normal-sodium diets (NSD-1.3% NaCl) from 8 weeks of age until the delivery of their first litter. After birth, the offspring received NSD. Tail-cuff blood pressure (TcBP) was measured in the offspring between 6 and 12 weeks of age. At 12 weeks of age, the offspring were subjected to either one week of HSD or low sodium diet (LSD-0.1 6%NaCl) feeding to evaluate RAAS responsiveness or to acute saline overload to examine sodium excretory function. Plasma (PRA) and renal renin content (RRC), serum aldosterone (ALDO) levels, and renal cortical and medullary renin mRNA expression levels were evaluated at the end of the study. Key findings: TcBP was higher among dams fed HSD, but no TcBP differences were observed among the offspring. Male offspring, however, exhibited increased TcBP after one week of HSD feeding, and this effect was independent of maternal diet. Increased RAAS responsiveness to the HSD and LSD was also observed in male offspring. The baseline levels of PRA. ALDO, and cortical and medullary renin gene expression were lower but the RRC levels were higher among HSD-fed male offspring (HSDoff). Conversely, female HSDoff showed reduced sodium excretion 4 h after saline overload compared with female NSDoff. Significance: High maternal sodium intake is associated with gender-specific changes in RAAS responsiveness among adult offspring. (C) 2012 Elsevier Inc. All rights reserved.

Enzyme activity in the small intestine of goat kids during the period of passive immunity acquisition

Enzyme activity of protein and carbohydrate degradation in small intestinal mucosa was investigated in goat kids fed with lyophilized bovine and goat colostrum. At 0,7 and 14 h of life 15 male newborns received 5% of body weight of lyophilized bovine colostrum and 14 goat colostrum, both with 55 mg/mL of IgG. Duodenum, jejunum and ileum samples were collected at 18,36 and 96 h of life. Three animals were sampled at birth, without colostrum intake. Activity of aminopeptidase N and A, dipeptidil peptidase IV, lactase, maltase and sucrase was determined as one international unit per gram of tissue. Intracellular enzymatic activity of acid phosphatase was observed by histochemistry in tissue section. Only the activity of aminopeptidase A in the ileum was affected by treatment, with a greater value for LBC than for GC (P < 0.05). The aminopeptidase N activity was the highest at 36 h in the duodenum (P < 0.05) and lowest at 96 h in the jejunum (P < 0.05). Dipeptidil peptidase IV activity was highest at 36 h in the duodenum (P < 0.05), lowest at 96 h in the jejunum (P < 0.05) and higher at 36 h than at 96 h in the ileum (P < 0.05). Aminopeptidase A activity in the ileum was highest at 36 h (P < 0.05), followed by 18 and 96 h of life (P < 0.05). Lactase activity in the duodenum increased from 18 to 36 h and from 36 to 96 h in the jejunum (P < 0.05). Maltase activity increased only in the duodenum from 18 to 96 h (P < 0.05). Sucrase activity in the jejunum decreased from 18 to 36 h and from 36 to 96 h in the ileum (P < 0.05). At birth, activity of most enzymes was similar to that at later times (P < 0.05). Histochemistry analyses showed a higher frequency of lysosomes with acid phosphatase activity in the duodenum, especially at 36 h of life. In the jejunum, the presence of lysosomes with acid phosphatase activity was the highest at 96 h, followed by 36 and 18 h of life. In the ileum, all samples showed low presence of lysosomes with acid phosphatase activity. These results indicate that lyophilized bovine colostrum, as a heterologous source of antibodies or nutrients, is a possible alternative management tool for goats. The present work also suggests that in the first 4 days of life, enzyme activity in the intestinal epithelium of goats is still not fully stimulated, which is an important characteristic for these animals that depend on macromolecule absorption to acquire passive protection after birth. (C) 2012 Elsevier B.V. All rights reserved.

Fiber architecture in remodeled myocardium revealed with a quantitative diffusion CMR tractography framework and histological validation

Background: The study of myofiber reorganization in the remote zone after myocardial infarction has been performed in 2D. Microstructural reorganization in remodeled hearts, however, can only be fully appreciated by considering myofibers as continuous 3D entities. The aim of this study was therefore to develop a technique for quantitative 3D diffusion CMR tractography of the heart, and to apply this method to quantify fiber architecture in the remote zone of remodeled hearts. Methods: Diffusion Tensor CMR of normal human, sheep, and rat hearts, as well as infarcted sheep hearts was performed ex vivo. Fiber tracts were generated with a fourth-order Runge-Kutta integration technique and classified statistically by the median, mean, maximum, or minimum helix angle (HA) along the tract. An index of tract coherence was derived from the relationship between these HA statistics. Histological validation was performed using phase-contrast microscopy. Results: In normal hearts, the subendocardial and subepicardial myofibers had a positive and negative HA, respectively, forming a symmetric distribution around the midmyocardium. However, in the remote zone of the infarcted hearts, a significant positive shift in HA was observed. The ratio between negative and positive HA variance was reduced from 0.96 +/- 0.16 in normal hearts to 0.22 +/- 0.08 in the remote zone of the remodeled hearts (p<0.05). This was confirmed histologically by the reduction of HA in the subepicardium from -52.03 degrees +/- 2.94 degrees in normal hearts to -37.48 degrees +/- 4.05 degrees in the remote zone of the remodeled hearts (p < 0.05). Conclusions: A significant reorganization of the 3D fiber continuum is observed in the remote zone of remodeled hearts. The positive (rightward) shift in HA in the remote zone is greatest in the subepicardium, but involves all layers of the myocardium. Tractography-based quantification, performed here for the first time in remodeled hearts, may provide a framework for assessing regional changes in the left ventricle following infarction.

Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil

Santos C.S.A.B., Piatti R.M., Azevedo S.S., Alves C.J., Higino S.S.S., Silva M.L.C.R., Brasil A.W.L. & Gennari S.M. 2012. Seroprevalence and risk factors associated with Chlamydophila abortus infection in dairy goats in the Northeast of Brazil. Pesquisa Veterinaria Brasileira 32(11):1082-1086. Unidade Academica de Medicina Veterinaria, Centro de Sa de e Tecnologia Rural, Universidade Federal de Campina Grande, Av. Universitaria s/n, Bairro Santa Cecilia, Patos, PB 58700-970, Brazil. E-mail: sergio.azevedo@pq.cnpq.br Few data are available on the prevalence and risk factors of Chlamydophila abortus infection in goats in Brazil. A cross-sectional study was carried out to determine the flock-level prevalence of C. abortus infection in goats from the semiarid region of the Paraiba State, Northeast region of Brazil, as well as to identify risk factors associated with the infection. Flocks were randomly selected and a pre-established number of female goats >= 12 mo old were sampled in each of these flocks. A total of 975 serum samples from 110 flocks were collected, and structured questionnaire focusing on risk factors for C. abortus infection was given to each farmer at the time of blood collection. For the serological diagnosis the complement fixation test (CFT) using C. abortus S26/3 strain as antigen was performed. The flock-level factors for C. abortus prevalence were tested using multivariate logistic regression model. Fifty-five flocks out of 110 presented at least one seropositive animal with an overall prevalence of 50.0% (95%; CI: 40.3%, 59.7%). Ninety-one out of 975 dairy goats examined were seropositive with titers >= 32, resulting in a frequency of 9.3%. Lend buck for breeding (odds ratio = 2.35; 95% CI: 1.04-5.33) and history of abortions (odds ratio = 3.06; 95% CI: 1.37-6.80) were associated with increased flock prevalence.

Sinais clínicos e ocorrência de anticorpos anti-Chlamydophila abortus em ovinos de São Paulo e Minas Gerais

A Chlamydophila abortusi, anteriormente conhecida como Chlamydia psittaci sovovar 1, é uma bactéria Gram negativa, intracelular obrigatória. Esse micro-organismo é frequentemente encontrado em distúrbios reprodutivos em ovinos, bovinos e caprinos, sendo o aborto epizoótico dos bovinos e o aborto enzoótico dos ovinos e caprinos as manifestações mais importantes. Considerando-se o pouco material literário a respeito da clamidofilose no Brasil, a pesquisa teve como objetivo determinar a presença de anticorpos fixadores de complemento anti-Chlamydophila abortusi, correlacionando os resultados obtidos com achados no exame clínico e histórico dos animais, além de alterações nos índices zootécnicos, em especial na esfera reprodutiva, tais como alto índice de repetição de cio, número elevado de abortamentos, elevado número de natimortos, entre outros. Foram testadas para prova de fixação do complemento 220 amostras de soro de ovinos, de 26 propriedades, distribuídas em 19 municípios, com relato de manifestação reprodutiva, obtendo-se 19,55% (43/220) de testes positivos para Chlamydophila abortusi, com ocorrência de foco constatada de 61,53%. No geral, a titulação de anticorpos encontrada foi baixa, com título não superior a 64. A frequência de manifestação reprodutiva mais observada foi o aborto, representando 65,12% (28/43) do número total de animais soropositivos, seguido de repetição de cio juntamente com nascimento de cordeiro fraco, com frequência de 6,98% (3/43) e, por fim, morte neonatal com 4,65% (2/43), sendo que não houve associação significativa entre animais que foram positivos ao teste e a esses fatores.

Sinais clínicos e ocorrência de anticorpos anti-Chlamydophila abortus em ovinos de São Paulo e Minas Gerais

A Chlamydophila abortus, anteriormente conhecida como Chlamydia psittaci sovovar 1, é uma bactéria Gram negativa, intracelular obrigatória. Esse micro-organismo é frequentemente encontrado em distúrbios reprodutivos em ovinos, bovinos e caprinos, sendo o aborto epizoótico dos bovinos e o aborto enzoótico dos ovinos e caprinos as manifestações mais importantes. Considerando-se o pouco material literário a respeito da clamidofilose no Brasil, a pesquisa teve como objetivo determinar a presença de anticorpos fixadores de complemento anti-Chlamydophila abortus, correlacionando os resultados obtidos com achados no exame clínico e histórico dos animais, além de alterações nos índices zootécnicos, em especial na esfera reprodutiva, tais como alto índice de repetição de cio, número elevado de abortamentos, elevado número de natimortos, entre outros. Foram testadas para prova de fixação do complemento 220 amostras de soro de ovinos, de 26 propriedades, distribuídas em 19 municípios, com relato de manifestação reprodutiva, obtendo-se 19,55% (43/220) de testes positivos para Chlamydophila abortus, com ocorrência de foco constatada de 61,53%. No geral, a titulação de anticorpos encontrada foi baixa, com título não superior a 64. A frequência de manifestação reprodutiva mais observada foi o aborto, representando 65,12% (28/43) do número total de animais soropositivos, seguido de repetição de cio juntamente com nascimento de cordeiro fraco, com frequência de 6,98% (3/ 43) e, por fim, morte neonatal com 4,65% (2/43), sendo que não houve associação significativa entre animais que foram positivos ao teste e a esses fatores.