Molecular tools and analytical approaches for the characterization of farm animal genetic diversity

Genetic studies of livestock populations focus on questions of domestication, within- and among-breed diversity, breed history and adaptive variation. In this review, we describe the use of different molecular markers and methods for data analysis used to address these questions. There is a clear trend towards the use of single nucleotide polymorphisms and whole-genome sequence information, the application of Bayesian or Approximate Bayesian analysis and the use of adaptive next to neutral diversity to support decisions on conservation.

Escherichia coli phylogenetic group determination and its application in the identification of the major animal source of fecal contamination

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

Refining the intestinal motility test in mice to reduce animal stress

A new protocol using 3-h fast animal for intestinal motility test was developed in our laboratory aiming the 3R's concept to reduce the stress of animals. Our results may aid in formulating recommendations that can be included in revised guidelines with regard to fasting time of mice.

Princípios éticos na experimentação animal

The treatment of animals used in research or in education tests requires proper care, scientific judgment and qualified professionals to understand the needs of the animals and the special requirements of the research, tests, and educational programs. The established guidelines aim the development of knowledge necessary for the improvement of health and wellbeing of humans as well as animals to get precise and accurate results. The techniques should not be performed with inappropriate procedures that cause pain or suffering. Some principles should be considered to have an appropriate animal care and treatment.

Estimates of covariance functions for growth of Nelore cattle applying a parametric correlation structure to model within-animal correlations

A total of 20,065 weights recorded on 3016 Nelore animals were used to estimate covariance functions for growth from birth to 630 days of age, assuming a parametric correlation structure to model within-animal correlations. The model of analysis included fixed effects of contemporary groups and age of dam as quadratic covariable. Mean trends were taken into account by a cubic regression on orthogonal polynomials of animal age. Genetic effects of the animal and its dam and maternal permanent environmental effects were modelled by random regressions on Legendre polynomials of age at recording. Changes in direct permanent environmental effect variances were modelled by a polynomial variance function, together with a parametric correlation function to account for correlations between ages. Stationary and nonstationary models were used to model within-animal correlations between different ages. Residual variances were considered homogeneous or heterogeneous, with changes modelled by a step or polynomial function of age at recording. Based on Bayesian information criterion, a model with a cubic variance function combined with a nonstationary correlation function for permanent environmental effects, with 49 parameters to be estimated, fitted best. Modelling within-animal correlations through a parametric correlation structure can describe the variation pattern adequately. Moreover, the number of parameters to be estimated can be decreased substantially compared to a model fitting random regression on Legendre polynomial of age. © 2004 Elsevier B.V. All rights reserved.

Water-soluble extract of Coleus barbatus modulates weight gain, energy utilization and lipid metabolism in secondary biliary cirrhosis: an experimental study in young rats.

PURPOSE: To test if a water extract of Coleus barbatus (WEB) has any effect on weight gain, food energy utilization and lipid metabolism in young rats with obstructive cholestasis. METHODS: Forty 21 day old (P21) Wistar rats, in groups of 10, were submitted to one of the following treatments: a sham operation with daily water or WEB administration, double ligature and resection of the bile duct with daily water or WEB administration. At P49 they were submitted for euthanasia when the following were determined: ingested feed (IF), energy utilization (EU) and weight gain (WG) from P29 to P49, together with total serum cholesterol (TC) and triacylglycerol (TG) concentrations, liver wet weight (LWW) and fat content (LFC). Two Way ANOVA and the S.N.K. test for paired comparisons were employed to study the effects of cholestasis and those of WEB and their interactions (p < or = 0.05). RESULTS: Cholestasis, independently of WEB, and WEB, independently of cholestasis both reduced IF, EU, and WG but there was no significant interaction between the two factors. Cholestasis, independently of WEB, increased LWW, LFC, the TC and TG The WEB, independently of cholestasis, reduced these values, and there was a significant interaction between the two factors; such that these effects were more accentuated in animals with cholestasis. CONCLUSION: The WEB reduced IF, WG, and EU, both in the presence and absence of cholestasis in the same proportion. It also partially inhibited the increase in LWW, LFC, TC and TG caused by cholestasis.

Fluorescence spectroscopy for tongue carcinoma detection: Study in an animal model

Fluorescence diagnosis of malignant lesions has been showed as an attractive optical technique due especially to its real-time response and a more objective and quantitative evaluation. Even though the oral cavity allows a direct examination many lesions are diagnosed when it is already in advanced stage, compromising the patient prognosis. In this study, the fluorescence spectroscopy was used to the detection of chemically induced carcinoma at the lateral border of the tongue in a hamster model. Two excitations wavelengths in visible region were applied: 442 and 532 nm. All the spectra results were analyzed comparing with the histopathological diagnosis. The better results were achieved with the 442 nm laser excitation. The spectra from carcinoma showed new emission bands and these were used to determined different ratios for a quantitative analysis. Using the 625-645 nm fluorescence range under 442 nm excitation (A3 coefficient) the percentage of false negative was of 9.1%, however the false positive percentage was of 18.5%. The 532 nm excitation provided a better normal tissue detection compared to 442 nm excitation. The ideal clinical condition is probably the use of multiple wavelengths excitation for a broader tissue fluorescence investigation.

Evaluation of serum- and animal protein-free media for the production of infectious bronchitis virus (M41) strain in a continuous cell line

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo retrospectivo de osteossarcoma primário dos ossos da pelve em cães em um período de 14 meses

The osteosarcoma (OSA) is one of the most common malignant tumors of dogs. Large and giant-breed dogs are affected and the mean age of these dogs is about 7 years. The tumor usually involves the metaphyses of long bones and the treatment is radical amputation and chemotherapy with cisplatin. The therapy of neoplasia located in pelvis is difficult and usually don't accept by owners. This report describes the incidence of primary osteosarcoma in pelvis of dogs, during a period of fourteen months. Of the eight animals taken care with this neoplasia, a bigger frequency in the dogs of the race Rottweiler and German Pastor was observed. Amongst the bones that compose animal's pelvis was observed greater osteosarcoma incidence in ileum (6 cases). Lameness, functional impotence of pelvic member and pain had been the main observed clinical signals. In seven cases was found advanced involvement osseous, mainly, for lyses of the cortical. All the animals had been submitted to the euthanasia for not presenting possibility of surgical treatment with safety margin. Also one was taken in consideration for this decision to the clinical condition of the patients, the possibilities to confer improvement in the quality of life of the dogs and the option of the owners' front to the prognostic.

The use of planarians as in vivo animal model to study laser biomodulation effects

A variety of effects is attributed to the photo stimulation of tissues, such as improved healing of ulcers, analgesic and anti-inflammatory effects, stimulation of the proliferation of cells of different origins and stimulation of bone repair. Some investigations that make qualitative evaluations, like wound healing and evaluation of pain and edema, can be conducted in human subjects. However, deeper investigations on the mechanisms of action of the light stimulus and other quantitative works that requires biopsies or destructive analysis has to be carried out in animal models or in cell cultures. In this work, we propose the use of planarians as a model to study laser-tissue interaction. Contrasting with cell cultures and unicellular organisms, planarians are among the simplest organism having tissue layers, central nerve system, digestive and excretory system that might have been platforms for the evolution of the complex and highly organized tissues and organs found in higher organisms. For the present study, 685 nm laser radiation was employed. Planarians were cut transversally, in a plane posterior to the auricles. The body fragments were left to regenerate and the proliferation dynamics of stem cells was studied by using histological analysis. Maximum cell count was obtained for the laser treated group at the 4th experimental day. At that experimental time, we also had the largest difference between the irradiated and the non-irradiated control group. We concluded that the studied flatworm could be an interesting animal model for in vivo studies of laser-tissue interactions.

Experimental model of pulmonary carcinogenesis in Wistar rats

Purpose: To elaborate an experimental model of pulmonary carcinogenesis in Wistar rats. Methods: Male Rattus norvegicus albinus, Wistar lineage was carried through an intra-pulmonary instillation of the Benzo[a]pyrene (B[a]P) dilution in alcohol 70%, a polycyclic aromatic hydrocarbon widely known by its power of tumoral induction. Three experimental groups had been formed with 08 animals each: Control Group (Alcohol 70%); B[a]P Group 10 mg/kg; e B[a]P Group 20mg/ kg, submitted to euthanasia 08, 10, 12 and 14 weeks after the experimental procedure. The pulmonary sections had been colored by hematoxilin-eosin (HE) and submitted to the morphometrical analysis to describe the tissue alterations. Results: The presence of diffuse inflammatory alterations was observed in all groups, however, at the analysis of the pulmonary tissue of the experimental groups, it had been observed hyperplasic alterations (BALT hyperplasia), and in one of the animals of the experimental group 20mg/kg (12 weeks), it was noticed the presence of cellular epithelial tracheal pleomorphism, suggesting the adenocarcinoma formation in situ. Conclusion: The main secondary alterations to the intra-pulmonary instillation of B[a]P in Wistar rats were: cellular proliferation, inflammatory alterations of several degrees and nodular lymphoid hyperplasias. The association of an activator agent of the pulmonary metabolic reply is necessary to establish the ideal reply-dose to the development of the lung cancer.

Ações de vigilância continuada, papel do cão como animal sentinela para toxoplasmose.

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii and its definitive host is the domestic and wild felids infecting human beings and other warmblooded animals. Dogs are considered a potential risk on the transmission due they can mechanically transmit oocysts to man. In this study, a retrospective analysis of toxoplasmic infection in dog serum samples sent to Serviço de Diagnóstico de Zoonoses/FMVZ-UNESP/Botucatu, SP, in the period of 1998 to 2007 was performed. During this period 1097 serum samples were analyzed by the indirect fluorescent antibody test (IFAT), with 299 (27.25%) positive. The most frequent titer was 16 (42.80%), followed by 64 (37.79%). The results indicate that T.gondii is distributed in the environment showing the role of the dog as sentinel animal to toxoplasmosis to monitor public health actions to the control of this zoonosis.

Avaliação clínica e parasitológica de cães naturalmente infectados por Leishmania (Leishmania) chagasi submetidos a tratamento com antimoniato de meglumina e alopurinol

Aiming to assess the efficacy of the treatment, to verify the occurrence of possible disease relapses and to search for the presence of parasites after the treatment, seven dogs naturally infected by Leishmania sp., were submitted to a treatment with meglumine antimoniate and allopurinol. For this, lymph node and bone marrow aspiration biopsies were carried out at seven moments. After the end of the six-month observation period all dogs were submitted to euthanasia. Then, spleen and liver imprints and in vitro cultures were carried out to search for amastigote forms of the parasite. All animals presented remission of the symptoms and during all the observation period no dog presented relapse of the disease, although amastigote forms of the parasite were observed in two of the animals at the end of the experiment. Thus, it was possible to conclude that the treatment promotes clinical healing but it does not eliminate the parasites completely.