Effect of the particle size of maize, rice, and sorghum in extruded diets for dogs on starch gelatinization, digestibility, and the fecal concentration of fermentation products

The influence of rice, maize, and sorghum raw material particle size in extruded dry dog food on the digestibility of nutrients and energy and the fecal concentration of fermentation products was investigated. Three diets with similar nutrient compositions were formulated, each with 1 starch source. Before incorporation into diets, the cereals were ground into 3 different particle sizes (approximately 300, 450, and 600 mu m); therefore, a total of 9 diets were in a 3 x 3 factorial arrangement (3 cereals and 3 particle sizes). Fifty-four beagle dogs (12.0 +/- 0.1 kg BW) were randomly assigned to the diets, with 6 dogs per diet. The digestibility was measured with the chromium oxide method. The data were evaluated with ANOVA considering the carbohydrate source, grinding effect, and interactions. The means were compared with the Tukey test and polynomial contrasts (P < 0.05). With the same grinding procedure, rice was reduced to smaller particles than other cereals. The cereal mean geometric diameter (MGD) was directly related to starch gelatini-zation (SG) during extrusion. For rice diets, the MGD and SG did not change nutrient digestibility (P > 0.05); only GE digestibility was reduced at the largest MGD (P < 0.01). For maize and sorghum diets, the total tract apparent nutrient digestibility was reduced for foods with greater MGD and less SG (P < 0.01). A linear reduction in nutrient digestibility according to cereal particle size was observed for sorghum (r(2) < 0.72; P < 0.01). Higher concentrations of fecal total shortchain fatty acids (SCFA) were observed for sorghum diets (P < 0.05) than for other diets. The rice diets led to the production of feces with less lactate (P < 0.05). The increase in raw material MGD did not influence fecal SCFA for rice diets, but for the dogs fed maize and sorghum foods, an increase in propionate and butyrate concentrations were observed as MGD increased (P < 0.05). In conclusion, for dogs fed different particle sizes of the cereal starches in the extruded diets, the digestibility and fecal characteristics were affected, and this effect was ingredient dependent.

Gallus gallus domesticus are resistant to infection with Neospora caninum tachyzoites of the NC-1 strain

The aim of this study was to experimentally evaluate infection in Gallus gallus domesticus with Neospora caninum tachyzoites of the NC-1 strain. Experimental infection was conducted in 90-day-old chickens, embryonated eggs and bioassays in dogs. In the first experiment, poults were randomly divided into four groups. Groups I and II were provided feed with coccidiostat, whereas groups III and IV received feed without coccidiostat. When the poults from groups I and III reached 90 days of age, they received a subcutaneous inoculation of N. caninum. Once the hens entered their egg-laying period, during the following 30 days, the eggs were collected, identified, weighed and placed in an incubator. On the 70th day after inoculation, all animals, including the chicks, were euthanized. Tissue samples from the adult poultry and chicks were collected for histopathology, immunohistochemistry (IHC) and PCR. Brain tissue and pectoral muscle samples from infected birds were fed to two dogs. Notably, the average weight of the group III eggs was lower than that of the group IV eggs (p <0.05). No changes consistent with infection in adult poultry or chicks were detected by histopathology or IHC; moreover, no amplified parasite DNA was detected in the birds'tissues or dogs'feces. No dog eliminated oocysts. In the second experiment, the embryonated chicken eggs were inoculated with 1 x 10(2) N. caninum tachyzoites, on the 10th day of incubation, and chicks born from these eggs were housed in boxes suitable for the species and received commercial feed and distilled water ad libitum. On the 30th day after infection (DAI), the poultry were euthanized, and their organs were processed as described in experiment I. The amplification of parasite DNA was observed in the spleen and pectoral muscles of one of the birds. The ingestion of bird tissues by dogs did not result in oocyst elimination. These results indicate that the parasite may have been eliminated by the host and that the use of tachyzoites to induce chronic disease might be a poor source for hens. (C) 2014 Elsevier B.V. All rights reserved.

Sequential healing at implants with different configuration and modified surfaces: an experimental study in the dog

To evaluate the peri-implant soft and hard tissue adaptation at implants with different modified surfaces and configurations. Six Beagle dogs were used. Mandibular premolars and first molars were extracted bilaterally. After 3 months, full-thickness flaps were elevated, and two different types of trans-mucosal implants (ICX-Gold®, Medentis Medical GmbH, Dernau, Germany and SLActive®, Institute Straumann, Bern, Switzerland) and two different surfaces were randomly installed in the distal regions of one side of the mandible. Abutments were applied, and a nonsubmerged healing was allowed. After 1 month, the procedures were performed in the other side of the mandible, and after a further month, the animals were sacrificed, biopsies were collected, and ground sections prepared for histological examination. Similar results in marginal bone and soft tissues dimensions were observed after 1 month of healing at the two implant systems used, and no major changes could be observed after 2 months of healing. After 1 month, the percentage of new bone was 69.0% and 68.8% at ICX-Gold and SLActive surfaces, respectively. After 2 months, the percentage of new bone was 67.8% and 71.9% at ICX-Gold Medentis and SLActive surfaces, respectively. No statistically significant differences in osseointegration were found. The two implant systems used resulted in similar osseointegration after 1 and 2 months of healing.

Epidemiological aspects of dog bites considering biter dogs and victims

The aim of this study was to characterize dog bites using data on biter dogs and victims. An exploratory cross-sectional study was performed using 203 records of individuals who had attended in public health services in 2009 in the municipality of Araçatuba, São Paulo, Brazil, after they had been bitten by a dog. Over 70% (92/129) of the biter dogs were male and most of them (71%) received as a gift. Dog owners reported companionship as the main reason for acquiring the dog. The victims who were children were predominantly male, while the victims who were elderly were predominantly female. Most children were bitten on the head/neck, while adults were bitten on the hands/feet and lower limbs (p<0.0001). The owner of the dog was known in 83.2% of cases. However, rabies observation of the biter dog following the attack was only reported in 59.4% of cases. Situations involving aggression were related to dogs having escaped from their home (18.7%) or roaming free on the streets (17.0%). The analysis of biting dog characteristics using information obtained from dog bite victims and biting dog owners can help direct the medical treatment for dog bite victims. Moreover, concepts of responsible dog ownership can reduce the occurrence of bites.

Role of ATM and PTEN in prostatic carcinogenic dog prostate: validation of aCGH results

The canis lupus familiares is the only species besides human that spontaneously develop prostatic carcinoma (PCa). In addition, the metastatic sites are similar to those frequently reported in men. For these reasons, the dog is the best natural model to study the molecular mechanisms in PCa development providing a natural animal model for treatment by molecular targets. Previously, we investigated copy number alterations by arrayCGH (Canine Genome CGH Microarray 4x44K-G2519F, Agilent Technologies) in canine prostatic lesions: 3 benign prostatic hyperplasias (BPH), 4 proliferative inflammatory atrophies (PIA), and 14 PCa. Five histologically normal prostatic tissues were used as reference. Genomic alterations were evaluated using Genomic Workbench Standard Edition 5.0.14. This previous study revealed significant copy number losses of Atm and Pten exclusively in PCa. In the present study, ATM and PTEN immunoexpression were investigated using a tissue microarray (TMA) containing 149 canine prostatic paraffin-embedded lesions (BPH, PIA and PCa) collected from 67 animals. Immunohistochemical reactions were performed using the polyclonal rabbit antibody anti-PTEN (Santa Cruz Biotech, 1:50) and anti-ATM (Abcam, 1:50). The sections were developed with diaminobenzidine (DAB) and peroxidase. The immunohistochemical staining was assessed in each core by the distribution of positive cells for each antibody per lesion (score 1: <25% cells positive, 2: 26% to 50%, 3: being 51% and 75% and 4:> 75%) and intensity (1: weak, 2: moderate, 3: intense). Chi-square or Fisher exact test was used to determine the association between the categorical variables using GraphPad Prism 5 (GraphPad Software Inc., La Jolla, CA). Distribution of positive cells did not differ among lesions. PCa and PIA showed more samples with weak intensity for ATM when compared to normal prostatic tissue and BPH (PCa: p=0,032 and PIA: p=0,025). Benign prostatic hyperplasia and normal samples presented intense PTEN immunostaining than PCa (p=0,021) and PIA (p=0,0013). These results suggest that ATM and PTEN proteins expression in canine prostatic carcinoma are downregulated possibly by copy number losses. These findings are similar from those described in prostate carcinomas from human corroborating for the use of dogs as a natural model to study prostatic disease in men.

Undifferentiated pulmonary adenocarcinoma of clear cells associated to hypertrophic osteopathy in a dog

Background: Most of the primary pulmonary tumors in dogs are malignant and from epithelial origin, being bronchioalveolar tumors more prevalent. Adenocarcinoma of clear cells, however, is a very rare pulmonary tumor and its origin is still unknown. It is related to several clinical abnormalities, including hypertrophic osteopathy, an unusual paraneoplastic syndrome characterized by a periosteal reaction along the shaft of long bones. Because of the unusual presentation of the pulmonary adenocarcinoma, the aim of this study was to describe the radiographic, histopathological, and immunohistochemical fi ndings of a dog affl icted with hypertrophic osteopathy secondary to an undifferentiated pulmonary adenocarcinoma of clear cells. Case: A 12-year-old, 45 kg, not castrated male Great Dane dog was presented with painful swelling of all four limbs and moderate respiratory distress. Radiographic examination and computed tomography of the limbs showed palisade-like periosteal bone proliferation involving radius, ulna, femur, patella, tibia, fi bula, tarsus, metacarpal, metatarsal and digits, suggesting hypertrophic osteopathy. Radiographic examination and computed tomography of the lungs also showed a round mass well delimited localized in the right diaphragmatic lobe. A lobectomy of the right diaphragmatic lobe and partial lobectomy of accessory lobe were performed. A poorly differentiated clear squamous cell carcinoma was diagnosed by histological examination. An immune-panel of CK5/CK6, CK7, p63 and TTF-1 was used for immunophenotyping. Immunostaining was weakly positive for CK5/CK6 and negative to all others. Therefore, the diagnosis was poorly differentiated clear cell adenocarcinoma. The dog showed improvement in clinical signs seven days after surgery. One month postoperatively, radiographic examination of the limbs showed less intense periosteal reaction and initiation of bone remodeling. Discussion: Primary pulmonary tumors are considered very infrequent in small animals, but its true incidence rate is dif- fi cult to establish in animal populations. The histological origin of the tumor in the present case, as verifi ed in the literature, is not well established by histological analysis. In these situations, the immunohistochemistry panel may be useful. The modifi cation of the diagnosis between histological analysis and by immunohistochemistry, among other factors, might be due to transdifferentiation from one phenotype to another at various stages in the neoplastic process. The clear cell appearance observed in this case may be verifi ed in all types of carcinoma due to intracellular accumulation of glycogen, most of which is dissolved during the preparation of paraffi n sections. This uncommon neoplasm apparently did not infl uence the radiographic or tomographic fi ndings of the hypertrophic osteopathy in the present case. The frequency of metastases depends on the histological type of the tumor, being common in the pulmonary adenocarcinoma and usually to tracheobronchial lymph nodes and pulmonary parenchyma. Although in this case the imaging studies did not show metastases to other pulmonary lobes, the histological exams showed metastatic lesions that may be associated to the dog’s death after the surgery.

T-cell lymphoma in the tongue of a dog with cutaneous and striated forelimb muscle involvement

Background: Primary tongue tumors rarely affect dogs and correspond to 4% of tumors involving the oropharynx. Until now, primary tongue lymphoma had not been reported. However, lymphoma involvement in the skeletal muscle, although quite unusual, was described in the literature in four cases. Cutaneous lymphoma is another rare extranodal manifestation. The objective of this report is to describe a case of T immunophenotype lymphoma occurrence, whose manifestation is atypical, not only because it is situated in the tongue muscle but also because of the subsequent involvement of the striated musculature of the left forelimb and the skin, which showed unfavorable evolution. Case: A female seven-year-old mongrel was seen showing a regular lump in the base of the tongue, 3 cm in diameter, not ulcerated and of fi rm consistency, with halitosis as the only clinical sign of the disease. Incisional biopsy of the lump was performed and histopathology verifi ed that it was large cell lymphoma. The material was sent for immunohistochemical evaluation and was characterized as T immunophenotype lymphoma by positive CD3 and negative CD79a marking. The CHOP (cyclophosphamide, doxorubicin, vincristine and prednisone) chemotherapy protocol was established as treatment and after the fi rst chemotherapy session there was partial remission of the mass, measuring 2 cm in diameter. The lump, however, remained stable in the following sessions. Thirty days after the diagnosis of lymphoma, the animal began to show lameness of the left forelimb and swelling near the head of the left humerus. A muscle mass, fi rm in consistency, progressing fast, presented a signifi cant increase, just three weeks after its appearance. Two skin lesions, arcuate, erythematous and pruritic also appeared in the dorsocervical and ventral-abdominal region. Incisional biopsy of these lesions was performed and the histopathological diagnosis confi rmed muscle and cutaneous large cell lymphoma and immunophenotype compatible with T cells (positive CD3 and negative CD79a). Due to disease advance, even during chemotherapy, a rescue protocol of L-asparaginase administration followed by lomustine and prednisone was proposed. Even with the rescue protocol there was no remission of the tumors and the case was classifi ed as progressive. The animal of this report died after completing the fi rst cycle of chemotherapy protocol, with a survival of 92 days. Discussion: Despite the fact that clinical behavior of primary lymphoma in dogs’ skeletal muscle is unknown, it is believed that, as in humans, it can be associated with chronic infl ammation or neoplastic cell invasion by proximity of the tumor or metastasis, which could justify the dissemination of the lymphoma reported here from the tongue to other tissues. However, appearance of concurrent independent lymphomas cannot be ruled out. As observed in the three cases of primary muscular lymphoma, the dog of this report had low response to therapy and short survival. This report presents the fi rst case of lymphoma in tongue with subsequent skin and left forelimb skeletal muscle involvement described in the literature. The clinical outcome corroborates the aggressiveness of muscular lymphoma observed in the other reports and also suggests that both tongue and other skeletal muscle tumors should be included in the differential diagnosis of canine lymphoma.

Occurrence of Cryptosporidium (Apicomplexa, Cryptosporidiidae) in Crotalus durissus terrificus (Serpentes, Viperidae) in Brazil

The objective of the present study was to investigate the prevalence of Cryptosporidium (Apicomplexa, Cryptosporidiidae) in the snake Crotalus durissus terrificus (Serpentes, Viperidae). Fifty animals were evaluated for the presence of oocysts of Cryptosporidium sp. at the time of arrival and 30 and 60 days later. Intestinal washings with saline solution (1% body weight), fecal samples, and organ scrapings were collected during the study. Oocysts were concentrated by an ether-phosphate-buffered saline sedimentation technique and then separated by a density gradient centrifugation technique. Smears were made with the sediment and submitted to modified acid-fast and auramine-rhodamine staining. Cryptosporidium-positive smears were used as controls for the experimental findings. The overall prevalence of Cryptosporidium sp. oocysts was 14%. Among the positive snakes, oocysts were detected only in the intestinal washing in two specimens, only in the feces in four specimens, and in both materials at least once in one specimen. The positive snakes were predominantly from Santa Maria da Serra city State of São Paulo (57.1%). We also observed that all of the examinations that presented positive results were obtained at least 27 days after the capture of the animals.

Avaliação histológica da próstata de cães adultos sexualmente intactos

Avaliou-se histologicamente a próstata de 30 cães adultos e idosos sexualmente intactos que apresentavam ou não sintomatologia clínica de doença prostática, e verificou-se a incidência de possíveis alterações da glândula. Dentre as alterações encontradas, a hiperplasia prostática benigna constituiu o diagnóstico mais comum, 85,6% (n=24), seguida por prostatite crônica, 64,3% (n=18), displasia do epitélio glandular, 42,8% (n=12), atrofia do epitélio glandular, 39,3% (n=11), infiltrado inflamatório focal, 25% (n=7), dilatação glandular focal, 21,4% (n=6), prostatite aguda, 7,1% (n=2), metaplasia escamosa, 3,6%, (n=1), metástase de neoplasia sistêmica, 3,6% (n=1) e abscesso prostático, 3,6% (n=1). Como em muitos casos os cães são assintomáticos, ressalta-se a importância da realização rotineira de exames clínicos específicos, como o toque retal e a ultrassonografia, para o diagnóstico precoce e o tratamento das afecções prostáticas.

Imunorreatividade da p53 associada à ausência de mutações no gene TP53 em linfomas caninos

Sabendo-se da influência das mutações no gene TP53 no desenvolvimento das neoplasias e da discrepância entre os resultados obtidos pelas técnicas de sequenciamento e imunoistoquímica, esta pesquisa teve como objetivo relacionar a sequência do TP53 com a imunorreatividade da p53. Foram obtidas amostras de linfoma de 12 cães. O diagnóstico histopatológico foi determinado pela classificação de Kiel. O imunofenótipo e a imunomarcação da p53 foram determinados por imunoistoquímica. Para reação com a p53, utilizou-se anticorpo policlonal anti-p53 (CM1) na diluição de 1:500. A região do gene TP53 compreendida entre os exons quatro e nove foi amplificada por PCR e submetida ao sequenciamento. Apesar dos resultados obtidos pela imunoistoquímica, nenhuma mutação foi encontrada nas sequências analisadas. Conclui-se que a imunorreatividade da p53 pela imunoistoquímica não pode ser atribuída à presença de mutações no domínio central do gene TP53.

Prevalence and antimicrobial resistance patterns of methicillin-resistant staphylococci (MRS) isolated in a Veterinary Teaching Hospital in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Dietary copper absorption and excretion in three semi-terrestrial grapsoid crabs with different levels of terrestrial adaptation

Three species of phylogenetically related semi-terrestrial crabs (Superfamily Grapsoidea - Sesarma rectum, Goniopsis cruentata and Neohelice granulata (formerly: Chasmagnathus granulatus) with different degrees of terrestriality were studied to quantify the accumulation of copper (Cu) in hemolymph, gills, hepatopancreas and antennal gland, and its excretion through the faeces. These crabs were fed for 15 days practical diets containing 0 (A), 0.5 (B), 1.0 (C), and 1.5% (D) of added CuCl2 (corresponding to 0, 0.2, 0.5 and 0.7% of Cu2+, respectively). The amount of food ingested was directly proportional to the degree of terrestriality: S. rectum, the most terrestrial species, ate around 2-3 times more than the other crabs, whereas G. cruentata ate 1.5-2 times more than N. granulata, the least terrestrial. The amount of Cu excreted in the feces was proportional to Cu ingestion, and was 76.8% and 64.2% higher for Sesarma fed diet D compared to G. cruentata and N. granulata, respectively. Sesarma also displayed higher Cu concentration in the haemolymph, gills and antennal glands, but not in the hepatopancreas. A detoxifying mechanism followed by elimination was probably present at this last organ, preventing Cu accumulation. More terrestrial crabs, such as Sesarma, may accumulate more Cu in hemolymph and tissues, showing a correlation between metal accumulation and increased terrestriality. In this aspect, contaminated feed sources with Cu may have more impact in conservation of terrestrial crabs. (C) 2008 Elsevier B.V. All rights reserved.

Isolation of extraintestinal pathogenic Escherichia coli from diarrheic dogs and their antimicrobial resistance profile

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Digestibilidade e balanço de nitrogênio em ovinos alimentados à base de dietas com elevado teor de concentrado e níveis crescentes de polpa cítrica peletizada

O experimento foi realizado com o objetivo de determinar os coeficientes de digestibilidade, a ingestão de nutrientes e o balanço de nitrogênio de dietas com 20% de silagem de milho e 80% de concentrado e 0, 25, 40 e 55% de polpa cítrica na matéria seca total, substituindo o milho em grão. em delineamento de blocos ao acaso, foram utilizados oito cordeiros mestiços, com nove meses de idade e 34 kg de peso, mantidos em gaiolas individuais com dispositivo para separação de fezes e urina. O período de coleta de alimento, sobras, fezes e urina foi de seis dias e a quantidade de alimento oferecido correspondeu à média do consumido nos três últimos dias do período de adaptação. O experimento foi desenvolvido em duas fases, utilizando-se os mesmos animais, que foram distribuídos em blocos pelo peso. Os resultados foram analisados por regressão polinomial. Houve aumento linear nos coeficientes de digestibilidade aparente da proteína bruta e da fibra em detergente ácido, com o aumento da participação da polpa cítrica na dieta. Os demais coeficientes de digestibilidade não diferiram entre si e foram, respectivamente: 71,8; 73,3; 77,2; 63,5; e 72,2%; para matéria seca, matéria orgânica, extrato etéreo, fibra em detergente neutro e hemicelulose. O teor médio de nutrientes digestíveis totais foi de 72,39%. Houve aumento linear da ingestão de matéria seca e de nutrientes digestíveis totais por dia e por peso metabólico, com o aumento de polpa cítrica na dieta. O mesmo ocorreu com a quantidade de nitrogênio ingerido, absorvido, urinário e retido. O aumento da participação da polpa cítrica peletizada até 55% da matéria seca da dieta, substituindo o grão de milho, implicou em melhor aproveitamento da fibra e eficiência no metabolismo do nitrogênio.