Fine-scale genetic diversity of the Brazilian Pantaneiro horse breed adapted to flooded regions.

Among the animal species first introduced in Brazil during the country's discovery, horses (Equus caballus) stand out because of their evolutionary history and relationship with humans. Among the Brazilian horse breeds, the Pantaneiro draws attention due to its adaptative traits. Blood samples of 116 Pantaneiro horses were divided into six populations based on their sampling location, aiming to identify the existence of genetic structure and quantify genetic diversity within and between them. Populations were compared to elucidate genetic variability and differentiation better and assess the impact of Pantanal's natural geographic barriers on gene flow between populations. Data from the GGP Equine BeadChip (Geneseek-Neogen, 65.157 SNPs) was used to assess basic diversity parameters, genetic distance (FST), principal component analysis (PCA), and population structure (ADMIXTURE) for the sampled animals. Mantel test was also performed to investigate the correlation between the populations' genetic and geographic distances. Results showed high genetic variability in all populations, with elevated levels of admixture in their structure. High levels of admixture make it challenging to establish a racial pattern and, consequently, populations within the breed, being that only one of the populations differentiated itself from the others. No significant correlations between genetic and geographic distances were observed, indicating that environmental barriers did not hinder gene flow between populations, and neither farmers' selection practices might have change breed genetic composition significantly. Low genetic distance and similar heterozygosity values were observed among populations, suggesting strong genetic proximity and low differentiation. Thereby, the Pantaneiro breed does not exhibit genetic subpopulations and could be considered, for conservation purposes, a single big population in the Panatnal region. This study will support sampling strategies for National genebank.

The Program for Biodiversity Research in Brazil: The role of regional networks for biodiversity knowledge, dissemination, and conservation.

The Program for Biodiversity Research (PPBio) is an innovative program designed to integrate all biodiversity research stakeholders. Operating since 2004, it has installed long-term ecological research sites throughout Brazil and its logic has been applied in some other southern-hemisphere countries. The program supports all aspects of research necessary to understand biodiversity and the processes that affect it. There are presently 161 sampling sites (see some of them at Supplementary Appendix), most of which use a standardized methodology that allows comparisons across biomes and through time. To date, there are about 1200 publications associated with PPBio that cover topics ranging from natural history to genetics and species distributions. Most of the field data and metadata are available through PPBio web sites or DataONE. Metadata is available for researchers that intend to explore the different faces of Brazilian biodiversity spatio-temporal variation, as well as for managers intending to improve conservation strategies. The Program also fostered, directly and indirectly, local technical capacity building, and supported the training of hundreds of undergraduate and graduate students. The main challenge is maintaining the long-term funding necessary to understand biodiversity patterns and processes under pressure from global environmental changes.

Integration of georeferenced and genetic data for the management of biodiversity in sheep genetic resources in Brazil.

There are few animal germplasm/gene bank collections in Brazil, and basic studies are needed to attend the future internal and external demands from international partners. The aim of this work was to validate a "proof of concept" that integrates spatial (georeferenced data) and genetic data regarding the local of origin from 3518 DNA samples from 17 different genetic groups or breeds of sheep in the Brazilian Germplasm bank. Spatialisation shows that not all genetic groups have samples in the bank, and collection is concentrated in the conservation nuclei spread nationwide. Only 21% of states with a specific breed have samples in the gene bank. The mean number of animals sampled per collection was 32, while the mean distance travelled to collect samples was 262 km from the conservation nuclei. For example, the Brazilian Somali were only collected in the conservation nucleus in Ceará State. No samples were collected to date for the Cariri breed, which is recognised by the Brazilian Ministry of Agriculture. Only two farms and one breed in the bank are from the northern region. Of the 27 states, there are samples in the gene bank of sheep from 13, so several states have no samples, requiring collection from herds outside the official system of conservation to make sure that studies using this germplasm realised are not biased. Significant genetic differences are seen above 332 km, which should guide future sampling efforts. Suggestions are given for improving the quantity, quality and diversity of samples in the gene bank.

A subunit vaccine based on biodegradable microspheres carrying rHsp65 protein and KLK protects BALB/c mice against tuberculosis infection.

Of the hundreds of new tuberculosis (TB) vaccine candidates, some have therapeutic value in addition to their prophylactic properties. This is the case for the DNA vaccine encoding heat-shock protein 65 (DNAhsp65) from Mycobacterium leprae. However, there are concerns about the use of DNA vaccines in certain populations such as newborns and pregnant women. Thus, the optimization of vaccination strategies that circumvent this limitation is a priority. This study evaluated the efficacy of a single dose subunit vaccine based on recombinant Hsp65 protein against infection with M. tuberculosis H37Rv. The Hsp65 protein in this study was either associated or not with immunostimulants, and was encapsulated in biodegradable PLGA microspheres. Our results demonstrate that the protein was entrapped in microspheres of adequate diameter to be engulfed by phagocytes. Mice vaccinated with a single dose of Hsp65-microspheres or Hsp65+CpG-microspheres developed both humoral and cellular-specific immune responses. However, they did not protect mice against challenge with M. tuberculosis. By contrast, Hsp65+KLK-microspheres induced specific immune responses that reduced bacilli loads and minimized lung parenchyma damage. These data suggest that a subunit vaccine based on recombinant protein Hsp65 is feasible.

Modelagem da dinâmica de rebanho de bovino de corte por meio de cadeia de Markov / Modeling dynamics of the beef cattle herd through Markov chain

A estrutura etária do rebanho e a estratégia de descarte de matrizes são pontos importantes na avaliação da eficiência do sistema de produção de gado de corte. Neste trabalho foi monitorado o rebanho de cria, cujas matrizes eram mantidas ou descartadas com base em critério técnico, conforme a avaliação do desempenho reprodutivo. Para tanto, foi utilizada a cadeia de Markov com o objetivo de simular a probabilidade de descarte por idade, ao longo da vida, e de calcular a expectativa de vida da matriz no rebanho. Após a introdução da estação de monta, a pressão de descarte foi maior, com aumento da probabilidade de descarte e da diminuição da probabilidade de permanência da matriz no rebanho nas idades mais avançadas. Com a introdução de tecnologias e especialmente da monta controlada, o processo torna-se mais dinâmico com a identificação e descarte de novilhas e matrizes improdutivas, diminuindo a idade média das vacas no rebanho e de descarte das mesmas.

The determination of age structure in a herd and the culling strategy in dams are important points to evaluate efficiency of a beef cattle production system. A breeding herd was monitored, where cows were maintained or discard according to technical criterion. Using the Markov chain the culling probability due to age throughout the life time of the cow was simulated. Life expectancy of the dam in the herd was also calculated. After the breeding season, culling pressure was higher and culling probability increased while permanency probability in the herd for older cows decreased, this made the herd more dynamic.

Body indices for the pantaneiro horse / Índices corporais para o cavalo pantaneiro

Body index measurements on 2248 horses registered by the Brazilian Association of Pantaneiro Horse Breeders (ABCCP), in the period from 1972 to 2000 were analyzed. Data was analyzed using SAS and DFREML to calculate genetic parameters. The Dactyl Thoracic Index indicated that these animals are intermediary (not light horses but not suitable for traction), while the BARON and CREVAT index confirmed this result. The Chest Index presented animals that are good for speed as they have long legs but looking at the Body index the indication was that these horses are neither suitable for speed nor traction (intermediary). The analysis of other conformation indices indicated that, in general, these horses are medilines, show good speed and chest development, making them suitable for dealing with cattle and resistance for long treks. The environmental factor analysis verified that these horses have changed in recent years possibly due to selection by the breeder which may affect their adaptive capacity.

Foram estudados índices zootécnicos de 2.248 animais registrados na Associação Brasileira de Criadores do Cavalo Pantaneiro (ABCCP), no período compreendido entre 1972 e 2000. Os dados foram analisados usando o programa SAS e o DFREML para estimação de parâmetros genéticos. Na análise dos índices corporais, o índice dáctilo torácico (IDT) apresentou animais intermediários (nem leves e nem aptos para tração). O índice de BARON e CREVAT também confirmou este resultado. Na determinação do índice peitoral (IP), os animais foram considerados, em geral, favoráveis à velocidade por apresentarem membros longos. Já no índice corporal (IC), a maioria dos animais não foi apta à velocidade e nem à tração (mediolíneos). Através da análise dos índices de conformação verificou-se que, em geral, os Pantaneiros são mediolíneos, aptos para velocidade e possuem bom desenvolvimento torácico, proporcionando velocidade na lida do gado e resistência a longas caminhadas. Na análise dos fatores ambientais, verificou-se que nos últimos anos alguns índices têm sofrido alterações, provavelmente devido à seleção já exercida pelo criador, que pode afetar a capacidade de adaptação dos cavalos.

Prime-boost vaccination based on DNA and protein-loaded microspheres for tuberculosis prevention.

We evaluated the use of a vaccine formulation based on a mixture of two different PLGA microspheres, composed by faster and slower release profiles, containing DNA encoding hsp65 and the recombinant hsp65 protein, respectively, aiming to DNA priming and protein boost after a single dose vaccination. The combination of PLGA50:50 microspheres containing DNA-hsp65 and trehalose dimycolate (TDM) with PLGA75:25 microspheres containing recombinant hsp65 (prime-boost Me) was able to induce high levels of anti-hsp65 specific antibodies. The serum levels of these specific antibodies remained high during 90 days after vaccination, whereas the DNA Me formulation based only in DNA-hsp65 plus TDM-loaded microspheres was not able to sustain the high antibody levels during the same period. Production of IFN-gamma was significant in animals vaccinated with both formulations, while the prime-boost Me vaccinated mice sustained higher levels of this cytokine during all the evaluation period. Thus, prime-boost strategy by using biodegradable microspheres seems to be a promising strategy to stimulate long-lasting immune response.

É possível uma vacina gênica auxiliar no controle da tuberculose? / Could a DNA vaccine be useful in the control of tuberculosis?

Vacinas de DNA, ainda em fase de experimentação e testes clínicos, podem se tornar uma importante ferramenta de combate a doenças infecciosas para as quais, até hoje, não existe prevenção segura e eficaz, como a tuberculose. Nos últimos anos vários estudos têm sido dedicados ao desenvolvimento de vacinas de DNA que codificam proteínas de micobactérias, entre as quais destacam-se as que codificam o antígeno 85 (Ag 85) e a proteína de choque térmico de 65 kDa (hsp65). Estes dois antígenos foram os mais estudados apresentando resultados bastante satisfatórios em ensaios pré-clínicos e com grande volume de dados registrados na literatura. Além de proteger contra infecção experimental por Mycobacterium tuberculosis virulenta, a vacina DNA-hsp65 também apresenta atividade terapêutica, ou seja, é capaz de curar os animais previamente infectados, inclusive aqueles com bacilos resistentes a múltiplas drogas. Esta vacina, hoje em avaliação clínica no Brasil também para o tratamento de câncer, é capaz de induzir a produção de citocinas de padrão Th1 tal como IFN- interferon-gama, associadas ao controle da doença. Além disso, a vacina de DNA-hsp65 é capaz de estimular clones de células CD8 citotóxicos e CD4 que podem ser caracterizados como células de memória sendo responsáveis por conferir imunidade duradoura contra a infecção. Quando utilizada na terapia da infecção, a vacina de DNA-hsp65 faz com que haja uma mudança no padrão de resposta imune, induzindo a secreção de citocinas de padrão Th1 criando um ambiente favorável à erradicação do bacilo. Os resultados demonstram ainda que a via de administração e a formulação na qual a vacina é administrada exerce fundamental influência no padrão e duração da resposta imune desencadeada. O conjunto de resultados hoje disponíveis mostra que uma vacina de DNA contra a tuberculose contribuirá de maneira significativa no controle desta doença.

Vaccine adjuvant: it makes the difference.

The use of protein or DNA in vaccination process rather than living or attenuated microorganism, aims at the increase of the vaccines safety. However, in these cases, the use of adjuvant is frequently required to improve their immunogenicity. In this study, we show the importance of the adjuvant in a vaccine formulation. Vaccines for tuberculosis provide an instructive example, based on the mycobacterial 65 kDa heat shock protein (hsp65). The same antigen can elicit completely different patterns of immune response depending on how it is administered. Thus, the same antigen might or not protect mice from challenge with Mycobacterium tuberculosis, depending on the formulation. These data suggest that, despite the name, the adjuvant plays a fundamental role on the vaccination process.