Abstract The establishment of epigenetic marks during the reprogramming window is susceptible to environmental influences, and stimuli during this critical stage can cause altered DNA methylation in offspring. In a previous study, we found that low levels of sulphur and cobalt (low S/Co) in the diet offered to oocyte donors altered the DNA methylome of bovine embryos. However, due to the extensive epigenetic reprogramming that occurs during embryogenesis, we hypothesized that the different methylation regions (DMRs) identified in the blastocysts may not maintain in adulthood. Here, we aimed to characterize DMRs previously identified in embryos, in the blood and sperm of adult progenies of two groups of heifers (low S/Co and control). We used six bulls and characterized the DNA methylation levels of KDM2A, KDM5A, KMT2D, and DOT1L genes. Our results showed that all DMRs analysed in both groups and tissues were hypermethylated unlike that noticed in the embryonic methylome profiles. These results suggest that embryo DMRs were reprogrammed during the final stages of de novo methylation during embryogenesis or later in development. Therefore, due to the highly dynamic epigenetic state during early embryonic development, we suggest that is essential to validate the DMRs found in embryos in adult individuals.

Abstract In the present study, the complete characterization of cDNA and genomic sequences of IL-1β and IL-8, as well as the expression profile of these genes in the South American fish pacu (Piaractus mesopotamicus) is provided. The full-length pmIL-1β cDNA was composed of 1208 nucleotides that would produce a precursor peptide with 273 amino acid residues. A putative caspase-1 cleavage site, similar to what is found in mammalian IL-1β, was identified producing a mature peptide with a theoretical molecular weight of 17.21 kDa. The pmIL-8 cDNA sequence consisted of 1019 nucleotides which encoded a 95-amino acid protein with a theoretical molecular weight of 10.43 kDa that showed all typical CXC chemokine features, including a 20-residue signal peptide and four conserved cysteine residues. Constitutive mRNA expression was detected for both genes in the liver, head kidney, gill, intestine, skin and spleen. After a bacterial challenge, up-regulation was detected for both pmIL-1β and pmIL-8 in the spleen and head kidney at 12 h post-infection. At 24 h post-infection there was a decrease in the expression of both genes, with pmIL-8 showing a significant down-regulation in the liver and head kidney when compared to the control groups.

Abstract Cross-species hybridizations have been extensively used to generate animals and plants better suited for draft and food and fiber production since Roman times, and are still important in current agricultural practices with growing uses especially in aquaculture. Diagnostic tools based on marker panels with sufficient numbers of markers for accurate identification of cross-species hybrid individuals from intercrossed and backcrossed populations are increasingly necessary for practical, accurate species-purity certification and management of commercial broodstocks. Minimal numbers of di-allelic markers with species-specific alleles required to accurately identify hybrid individuals in intercrossed and advanced backcrossed populations were estimated using power analysis, and ranged from 5 to 191 (α = .05), and from 7 to 293 (α = .01), considering backcross 1 (BC1) to BC6 populations, respectively. Numbers of markers required for accurate hybrid identification observed in simulated BC1 to BC6 populations ranged from 5 to 1,131 and 7 to 8,065, considering error rates ≤ 5% and ≤ 1%, respectively. Estimated and observed numbers of diagnostic markers required for accurate hybrid identification up to four generations of backcrossing fall within practical operational limits of most commercial platforms currently available for genotyping low-density SNP marker panels. Therefore, cost-effective assay panels could be developed to provide practical tools for accurate species-purity certification.

Resumo: O objetivo deste trabalho foi avaliar a utilidade de um subconjunto de 18 polimorfismos de nucleotídeo único (SNPs) para a certificação das raças de ovinos Crioula Brasileira, Morada Nova (MN) e Santa Inês (SI). Dados de 588 animais foram analisados com o programa Structure. Em 82% dos casos, observou-se designação racial correta com confiança acima de 90%. A maioria dos casos de designação incorreta de raça foi observada em MN e SI. Portanto, apesar de o subconjunto de 18 SNPs ter confiabilidade elevada, ele não é suficiente para a inequívoca certificação das raças estudadas, principalmente das deslanadas. É necessário o desenvolvimento de um painel mais preciso para uso amplo em certificação racial.

Abstract: The objective of this work was to evaluate the usefulness of a subset of 18 single nucleotide polymorphisms (SNPs) for breed identification of Brazilian Crioula, Morada Nova (MN), and Santa Inês (SI) sheep. Data of 588 animals were analyzed with the Structure software. Assignments higher than 90% confidence were observed in 82% of the studied samples. Most of the low-value assignments were observed in MN and SI breeds. Therefore, although there is a high reliability in this subset of 18 SNPs, it is not enough for an unequivocal assignment of the studied breeds, mainly of hair breeds. A more precise panel still needs to be developed for the widespread use in breed assignment.

Resumo: As corridas em homozigose (ROHs) são longos trechos genômicos em homozigose, identificáveis por meio de marcadores moleculares, que podem fornecer informações genômicas para estimativas precisas, utilizadas para caracterizar populações, determinar história evolutiva e informações demográficas, estimar níveis de consanguinidade e identificar assinaturas de seleção em animais de interesse zootécnico. Este artigo de revisão tem por objetivo fazer um levantamento dos trabalhos sobre a eficiência de ROHs para essas finalidades. Fatores como deriva genética, seleção natural ou artificial, efeito fundador e número efetivo da população influenciam diretamente o tamanho e a distribuição das ROHs pelo genoma. Individualmente, a estimativa genômica de consanguinidade baseada em ROHs pode ser realizada por meio do índice FROH, que, em geral, é considerado mais preciso do que índices baseados em outros tipos de informações genômicas ou genealógicas. Altas frequências de ROHs específicas em uma população podem ser utilizadas para identificar assinaturas de seleção. Os resultados de trabalhos recentes com ROHs em animais domésticos têm mostrado a eficiência de seu uso para caracterização de rebanhos, de forma confiável e acessível, a partir de informações genômicas.

Abstract: Runs of homozygosity (ROHs) are long stretches of homozygous genomic segments, identifiable by molecular markers, which can provide genomic information for accurate estimates to characterize populations, determine evolutionary history and demographic information, estimate levels of consanguinity, and identify selection signatures in production animals. This review paper aims to perform a survey of the works on the efficiency of ROHs for these purposes. Factors such as genetic drift, natural or artificial selection, founder effect, and effective population size directly influence the size and distribution of ROHs along the genome. Individually, genome estimates of consanguinity based on ROHs can be obtained using the FROH index, which is generally considered more accurate than indexes based on other types of genomic or genealogical information. High frequencies of specific ROHs in a population can be used to identify selection signatures. The results of recent studies with ROHs in domestic animals have shown the efficiency of their use to characterize herds in a reliable and accessible way, using genomic information.

Resumo: O recente desenvolvimento de painéis de “single nucleotide polymorphisms” (SNPs) distribuídos ao longo do genoma possibilitou a realização de diversos trabalhos com diferentes espécies. O processo seletivo promove o aumento ou a diminuição da frequência alélica (ou gênica) em loci específicos do genoma, além de promover o arrasto dos alelos próximos no cromossomo. Desta forma, são formadas regiões do genoma com o aumento na frequência de determinados alelos na população, o que caracteriza a assinatura de seleção. A detecção destas assinaturas é importante para a caracterização dos recursos genéticos, bem como a identificação de genes ou regiões envolvidos no controle e na expressão de características de importância produtiva e econômica. Os ovinos são uma importante espécie para estes estudos, uma vez que encontram-se amplamente dispersos em diferentes ambientes e apresentam grande diversidade fenotípica. Devido à grande quantidade de dados gerados nas análises genômicas, são necessários métodos estatísticos e programas específicos para a detecção de assinaturas de seleção. Assim, os objetivos deste artigo de revisão são apresentar os principais métodos estatísticos e os programas atualmente utilizados para análise de dados genômicos e a detecção de assinaturas de seleção; descrever os resultados dos recentes trabalhos publicados sobre assinaturas de seleção em ovinos; e discutir alguns desafios e oportunidades nesta área de pesquisa.

Abstract: The recent development of genome-wide single nucleotide polymorphism (SNP) arrays made it possible to carry out several studies with different species. The selection process can increase or reduce allelic (or genic) frequencies at specific loci in the genome, besides dragging neighboring alleles in the chromosome. This way, genomic regions with increased frequencies of specific alleles are formed, caracterizing selection signatures or selective sweeps. The detection of these signatures is important to characterize genetic resources, as well as to identify genes or regions involved in the control and expression of important production and economic traits. Sheep are an important species for theses studies as they are dispersed worldwide and have great phenotypic diversity. Due to the large amounts of genomic data generated, specific statistical methods and softwares are necessary for the detection of selection signatures. Therefore, the objectives of this review are to address the main statistical methods and softwares currently used for the analysis of genomic data and the identification of selection signatures; to describe the results of recent works published on selection signatures in sheep; and to discuss some challenges and opportunities in this research field.

Resumo: O objetivo deste trabalho foi identificar polimorfismo de nucleotídeo único (SNPs) em dados de ressequenciamento dos genes MC1R, ASIP e TYRP1, obtidos de ovelhas Crioulas (Ovis aries) com cores de pelagem distintas. Polimorfismos nos genes ASIP (agouti-signaling protein), MC1R (melanocortin 1 receptor) e TYRP1 (tyrosinase-related protein 1) foram analisados em 115 ovinos, provenientes do núcleo de conservação da ovelha Crioula, da Embrapa. No total, 7.914 pb foram sequenciados por animal, e 14 SNPs foram identificados. Dois ensaios adicionais foram realizados para detectar duplicações e deleções no gene ASIP. Noventa e cinco por cento da variação de cor da pelagem foi explicada pelas interações epistáticas entre alelos específicos dos genes MC1R e ASIP. Evidências sugerem um importante papel das variantes TYRP1 para cor da lã, apesar das suas baixas frequências. O painel de marcadores usado foi eficiente em detectar alelos associados à coloração da pelagem nos animais estudados e, portanto, pode ser usado na implementação de um programa de seleção assistida por marcadores, no núcleo de conservação de ovinos da raça Crioula.

Abstract: The objective of this work was to identify single nucleotide polymorphisms (SNPs) in resequencing data from MC1R, ASIP, and TYRP1 genes derived from Crioula sheep (Ovis aris) with different coat colors. Polymorphisms in the ASIP (agouti-signaling protein), MC1R (melanocortin 1 receptor), and TRYP1 (tyrosinase-related protein 1) genes were analyzed in 115 sheep from Embrapa’s conservation nucleus of crioula sheep, in Brazil. A total of 7,914 bp were sequenced per animal, and 14 SNPs were identified. Two additional assays were performed to detect duplications and deletions in the ASIP gene. Ninety-five percent of the coat color variation was explained by epistatic interactions observed between specific alleles in the MC1R and ASIP genes. Evidence suggests an important role of TYRP1 variants for wool color, despite their low frequencies. The marker panel was efficient enough in predicting coat color in the studied animals and, therefore, can be used to implement a marker-assisted selection program in the conservation nucleus of sheep of the crioula breed.

RESUMO: A crescente evolução das tecnologias de sequenciamento e genotipagem de DNA tornaram possível analisar o genoma de várias espécies, compreender suas funções dentro dos sistemas biológicos e, sobretudo, começar a entender os mecanismos que controlam as interações entre os genótipos e os efeitos ambientais que estão envolvidos com a expressão de características de interesse econômico. Várias tecnologias foram apresentadas por diferentes empresas para a genotipagem de painéis de SNP de baixa densidade, os quais podem ser utilizados em diferentes aplicações com objetivos variados, desde testes de paternidade e diagnósticos de doenças genéticas até a identificação de animais geneticamente superiores, com base em polimorfismos caracterizados em genes candidatos. Essa revisão analisa a tecnologia Goldengate Beadxpress e coloca em perspectiva seu uso nessas aplicações.

ABSTRACT: The increasing development of DNA sequencing and genotyping technologies has made possible to analyze the genomes of several species. Genomic studies of production animals have greatly increased the understanding of mechanisms that control the interactions of genetic and environmental factors involved in the expression of traits of economic importance. Several technologies have been presented by different companies for the genotyping of low-density SNP panels, which may be used in different applications with different goals, such as paternity testing, diagnosis of genetic diseases, and identification of genetically superior animals based on polymorphisms characterized in candidate genes. The present review critically analyzes the GoldenGate Beadxpress technology and puts its use in these applications into perspective.

Brazilian goats are generally kept in small herds and extensive rearing systems, mainly in the northeastern region of the country. Despite production improvement in recent years, the lack of pedigree control has affected genetic progress. This study aimed to validate a panel of 16 microsatellites for parentage testing in locally adapted and commercial goats breeds raised in Brazil, as well as to compare its efficiency with the panel recommended by the Brazilian Ministry of Agriculture, Livestock and Supplies (MAPA) in 2004. The number of alleles and expected heterozygosity (He) per marker ranged from four to 18, and from 0.051 to 0.831, respectively. Using all markers, 100% of parentage cases of the validation dataset were resolved with a strict confidence level of 95%. The 16 microsatellites panel showed adequate exclusion power (99.99%) and identity accuracy (99.99%). Suggestions for improvement of the marker panel endorsed by MAPA are provided.

This study estimated the genetic diversity and structure of 12 genetic groups (GG) of locally adapted and specialized pigs in the state of Pernambuco using 22 microsatellite markers. Nine locally adapted breeds (Baé, Caruncho, Canastra, Canastrão, Mamelado, Moura, Nilo, Piau and UDB (Undefined Breed)) and 3 specialized breeds (Duroc, Landrace and Large White), totaling 190 animals, were analyzed. The Analysis of Molecular Variance (AMOVA) showed that 3.2% of the total variation was due to differences between genetic groups, and 3.6% to differences between local and commercial pigs. One hundred and ninety eight alleles were identified and apart from the Large White breed, all GG presented Hardy-Weinberg Equilibrium deviations for some loci. The total and effective allele means were lower for Duroc (3.65 and 3.01) and higher for UDB (8.89 and 4.53) and Canastra (8.61 and 4.58). Using Nei's standard genetic distance and the UPGMA method, it was possible to observe that the Landrace breed was grouped with the local genetic groups Canastra, Moura, Canastrão, Baé and Caruncho. Due to the complex admixture pattern, the genetic variability of the 12 genetic groups can be analyzed by distributing the individuals into two populations as demonstrated by a Bayesian analysis, corroborating the results from AMOVA, which revealed a low level of genetic differentiation between the inferred populations.

O objetivo deste trabalho foi investigar prováveis modos de ação da levedura Cryptococcus magnus, que resultam no controle da antracnose (Colletotrichum gloeosporioides) em frutos de mamoeiro na póscolheita. A microscopia eletrônica de varredura foi utilizada para avaliar o efeito da levedura sobre inoculações realizadas após a colheita. Os resultados mostraram que C. magnus é capaz de colonizar a superfície de ferimentos nos frutos e vencer a competição por espaço e, provavelmente, por nutrientes. Além disso, C. magnus produz uma matriz de textura característica que afeta a integridade da hifa do patógeno. A competição por espaço e a produção de substâncias que afetam a integridade das hifas estão entre os mais importantes modos de ação desta levedura.

The objective of this work was to investigate possible modes of action of the yeast Cryptococcus magnus in controlling anthracnose (Colletotrichum gloeosporioides) on post harvested papaya fruits. Scanning electron microscopy was used to analyze the effect of the yeast on inoculations done after harvest. Results showed that C. magnus is able to colonize wound surfaces much faster than the pathogen, outcompeting the later for space and probably for nutrients. In addition, C. magnus produces a flocculent matrix, which affects hyphae integrity. The competition for space and the production of substances that affect hyphae integrity are among the most important modes of action of this yeast.