C.G. Fonseca

Genetic diversity between herds of Alpine and Saanen dairy goats and the naturalized Brazilian Moxotó breed

Brazilian naturalized goat breeds are adapted to the semiarid conditions prevalent in the Northeast region of the country (which has the largest Brazilian goat heard) and represent an as yet uninvestigated source of genetic diversity. Currently, imported goat breeds are crossed with Brazilian naturalized goat breeds, endangering the genetic potential of the naturalized breeds. We used 11 microsatellite markers to determine the genetic diversity among imported (non-naturalized) dairy Alpine and Saanen goats and naturalized Brazilian Moxoto goats. We genotyped 292 goats from three herds (one private, one from the University of Minas Gerais and the Moxoto conservation herd from Embrapa Caprinos) and found that the general heterozygosity was 0.6952 for Alpine, 0.7043 for Saanen and 0.4984 for Moxoto goats. The number of alleles ranged from 5 (INRA005) to 11 (BM3205), with an average of 7 alleles per locus in the imported breeds and 3.5 alleles per locus in the Moxoto breed. Mean differentiation between populations was higher for herds (FSTS = 0.0768) than for breeds (FSTP = 0.0263), indicating similarity between the imported breeds and the existence of crosses between them. Neis genetic distance was highest between the Moxoto breed and the imported breeds. These indicate that further studies using these molecular markers would be fruitful.

Cryptic Speciation in Brazilian Epiperipatus (Onychophora: Peripatidae) Reveals an Underestimated Diversity among the Peripatid Velvet Worms

BACKGROUND Taxonomical studies of the neotropical Peripatidae (Onychophora, velvet worms) have proven difficult, due to intraspecific variation and uniformity of morphological characters across this onychophoran subgroup. We therefore used molecular approaches, in addition to morphological methods, to explore the diversity of Epiperipatus from the Minas Gerais State of Brazil. METHODOLOGY/PRINCIPAL FINDINGS Our analyses revealed three new species. While Epiperipatus diadenoproctussp. nov. can be distinguished from E. adenocryptussp. nov. and E. paurognostussp. nov. based on morphology and specific nucleotide positions in the mitochondrial cytochrome c oxidase subunit I (COI) and small ribosomal subunit RNA gene sequences (12S rRNA), anatomical differences between the two latter species are not evident. However, our phylogenetic analyses of molecular data suggest that they are cryptic species, with high Bayesian posterior probabilities and bootstrap and Bremer support values for each species clade. The sister group relationship of E. adenocryptussp. nov. and E. paurognostussp. nov. in our analyses correlates with the remarkable morphological similarity of these two species. To assess the species status of the new species, we performed a statistical parsimony network analysis based on 582 base pairs of the COI gene in our specimens, with the connection probability set to 95%. Our findings revealed no connections between groups of haplotypes, which have been recognized as allopatric lineages in our phylogenetic analyses, thus supporting our suggestion that they are separate species. CONCLUSIONS/SIGNIFICANCE Our findings suggest high cryptic species diversity and endemism among the neotropical Peripatidae and demonstrate that the combination of morphological and molecular approaches is helpful for clarifying the taxonomy and species diversity of this apparently large and diverse onychophoran group.

Variations in genotype-phenotype correlations in phenylketonuria patients.

Phenylalanine hydroxylase deficiency is a trait inherited in an autosomal recessive pattern; the associated phenotype varies considerably. This variation is mainly due to the considerable allelic heterogeneity in the phenylalanine hydroxylase enzyme locus. We examined the genotype-phenotype correlation in 54 phenylketonuria (PKU) patients from Minas Gerais, Brazil. Two systems were used. The first was a phenotype prediction system based on arbitrary values (AV) attributed to each mutation and the second was a correlation analysis. An AV was assigned to each mutation: AV = 1 for classical PKU mutation; AV = 2 for moderate PKU mutation; AV = 4 for mild PKU mutation, and AV = 8 for non-PKU hyperphenylalaninemia mutation. The observed phenotype for AV analysis was the clinical diagnosis established by the overloading phenylalanine test. Among the 51 PKU patients that we analyzed based on this trait, in 51% the predicted phenotype did not match the observed phenotype; the highest degree of concordance was found in patients with null/null genotypes. The genotype was observed to be a good predictor of the clinical course of the patients and significant correlations were found between phenylalanine values at first interview and predicted residual activity, genotype and arbitrary value sum.

Análise temporal da endogamia e do tamanho efetivo da população de eqüinos da raça Mangalarga Marchador

A partir de informacoes de 286.047 animais registrados na Associacao Brasileira dos Criadores do Cavalo Mangalarga Marchador, desde a sua fundacao, em 1949, ate dezembro de 1999, verificaram-se o coeficiente de endogamia e o tamanho efetivo da raca Mangalarga Marchador. A media do coeficiente de endogamia para toda a populacao foi de 1,3% e diferente de zero para 22,6% dos animais. Considerando apenas os animais endogâmicos, o coeficiente medio de endogamia foi de 5,7%, minimo de 0,001 e maximo de 46,9%. Observou-se que 50% da populacao endogâmica apresentou coeficiente de endogamia entre 0,0001 e 10%. Na populacao atual a media de endogamia foi 3,8%, enquanto a media nos pais foi de 7,3%. O tamanho efetivo da populacao variou entre os periodos bianuais de nascimentos, sendo de 9.174,24 animais para o periodo de 1998-1999. O valor maximo para a razao entre o tamanho efetivo e o numero de animais da populacao foi quase sempre inferior a 0,50, variando de 0,39 em 1980-1981 a 0,79 em 1954-1955.

Brief communication: Admixture analysis with forensic microsatellites in Minas Gerais, Brazil: the ongoing evolution of the capital and of an African-derived community.

We report the estimated allele frequencies for 13 and 14 microsatellite loci in two populations of Minas Gerais, Brazil as follows: Belo Horizonte (the capital) and Marinhos (an African-derived community). Analysis of the African, Amerindian, and European genetic contributions to both populations, together with historical information, revealed distinct differences between the two populations. Estimates for Belo Horizonte revealed a higher-European (66%) than African (32%) contribution, and a minimal Amerindian contribution. These results are consistent with the peopling of the city mainly by people from the Minas Gerais hinterland, a people highly admixed but with more European ancestry. Estimates for Marinhos confirmed the high-African component of the population. However, a temporal analysis of two datasets-CURRENT (representing the population living in Marinhos today) and ORIGINAL (representing families, who have lived in Marinhos since the onset of the 20th century),-identified a diminishing of the populations African ancestry from 92% in the ORIGINAL group to 67% in the CURRENT group. This change is here interpreted as a consequence of the growing migration into the village of people with more European ancestry and subsequent admixture with the local population.

PKU in Minas Gerais State, Brazil: Mutation Analysis

This work was undertaken in order to ascertain the PKU mutational spectrum in Minas Gerais, Brazil, the relative frequency of the mutations in the State and the origin of these mutations by haplotype determination. Minas Gerais is a trihybrid population formed by miscegenation from Europeans, Africans and Amerindians. All 13 exons of the PAH gene from 78 PKU patients were analyzed, including splicing sites and the promoter region. We identified 30 different mutations and 98% of the PAH alleles were established. A new mutation (Q267X) was identified as well. The most common mutations found were V388M (21.2), R261Q (16.0%), IVS10‐11G>A (15.3%), I65T (5.8%), IVS2+5G>C (5.8%), R252W (5.1%), IVS2+5G>A (4.5%), P281L (3.8%) and L348V (3.2%). These nine mutations correspond to 80% of the PKU alleles in the state. Haplotypes were determined to characterize the origin of the PAH alleles. The majority of the mutations found, with respective haplotypes, are frequent in the Iberian Peninsula. However, there were some mutations that are rare in Europe and four previously unreported mutation‐haplotype associations. I65T and Q267X were found in association with haplotype 38 and may be African in origin or the result of miscegenation in the Brazilian population.

Genetic diversity and population structure in Brazilian Mangalarga Marchador horses.

One hundred and fifteen unrelated Mangalarga Marchador horses were sampled from three geographically distinct regions of Minas Gerais State, Brazil (South, Southeast, and Northeast) and tested for 10 microsatellite loci. Genetic diversity and population structure parameters were estimated with ARLEQUIN 3.0, CERVUS 2.0, POPGENE 1.31, GENEPOP on the web, STRUCTURE 2.0, and SPAGEDI 1.2 software packages. Under Hardy-Weinberg assumptions, seven markers were at equilibrium (LEX014, LEX017, LEX019, SGCV23, TKY321, VHL20, and VIASH39), while two (ASB3 and LEX031) presented significant homozygote excess. Seventy-four alleles were identified in these nine markers, with a mean of 8.22 alleles. Mean heterozygosity was 0.637 and polymorphism information content was 0.662. Markers ASB3, LEX019, SGCV23, TKY321, and VHL20 were highly informative (PIC >0.7) and may be useful for eventual expansion of parentage test panels. The F(ST) value (0.0562) indicated relatively little geographical structure. However, based on a Bayesian-based cluster analysis under a three-cluster model, 94% of the 115 individuals were correctly assigned to the subpopulations from where they were sampled. Mean pairwise f was relatively high (0.11), and in spite of the efforts towards non-consanguineous sampling, 1% of the pairs of individuals shared over 50% of the alleles. These results strongly suggest that the population is genetically structured. Under a conservation genetics approach, two strategies are recommended: avoidance of crosses between highly endogamic individuals and stimulation of crosses between individuals from those regions for which low genetic flow was identified.

Delineating geographic boundaries of the woolly mouse opossums, Micoureus demerarae and Micoureus paraguayanus (Didelphimorphia: Didelphidae)

This paper reports on molecular classification of the woolly mouse opossum, Micoureus spp., in the southeastern Atlantic Forest in Brazil, a hotspot of critically threatened biodiversity. Phylogenetic analysis and character-based diagnosis were done using DNA sequences from the mitochondrial cytochrome b gene and cytochrome c oxidase subunit I genes, and exon 6 of the nuclear dentine matrix protein 1 gene (DMP1). Although the nuclear DMP1 gene showed insufficient genetic variation for species diagnosis, the mtDNA analyses resulted in the robust grouping of samples of the M. paraguayanus/M. demerarae complex into three clades with distinct DNA sequence diagnostics for the species units in this study. The results support the species status of M. paraguayanus (Tate in Am Mus Novit 493: 1–13, 1931), which has a geographic distribution in the Atlantic Forest from the North and Northeast of Minas Gerais state in Brazil, going south along the coastal region of Brazil, to Paraguay and Argentina. Evidence of the boundary for this species and the provided diagnostics should facilitate and improve accuracy of studies that have been done in critical threatened fragments of the Atlantic Forest, especially in Minas Gerais and Bahia states, Brazil.

Análise multivariada de resultados da ovulação múltipla seguida de transferência de embriões de doadoras zebuínas

Data on 1390 Zebu donors multiple ovulation followed by embryo transfer results were analyzed. The donors came from different regions of Brazil and belonged to Brahman (31), Gir (22), Guzera (68) and Nelore (411) breeds. The age of donors ranged from 1.5 to 19.5 years old. The first recovery data and consecutive recovery data were submitted to multivariate analysis. The estimated Pearson correlation coefficients were: 0.72 between total recovered structures (ER) and number of viable embryos (EV); 0.66 between ER and total of positive pregnant receptors (PP); 0.53 between ER and number of palpated corpora luteum; 0.89 between EV and PP; 0.33 between EV and CL and 0.30 between PP and CL. Relationships among inbreeding coefficient, age of donors to superovulation, stimulating drug, drug dosage, recovery number and number of executed insemination were also observed. Factor analysis indicated the following factors for consecutive recoveries: factor 1 - factors that reduce superovulation response, factor 2 - superovulation scheme according to characteristics of donors, factor 3 - success of superovulation scheme. For first recovery analysis the following factors were designated: factor 1 - optimum age for superovulation, factor 2 - superovulation scheme according to characteristics of donors, and factor 3 - alternative superovulation scheme. The analysis also indicated the results of superovulation are quite variable and that the age of donors to superovulation, inbreeding coefficient of donors, recovery number, drug dosage and number of inseminations have high effect on the results. The number of viable embryos and the correlation between number of viable embryos and total of positive pregnant receptors can be indicators of MOET success.

PERMANENT GENETIC RESOURCES: Characterization of eight microsatellite loci in the woolly mouse opossum, Micoureus paraguayanus, isolated from Micoureus demerarae

Eight novel microsatellite markers were isolated from the woolly mouse opossum from the Amazon Forest in Peru, Micoureus demerarae, using a partial genomic DNA library and an enrichment protocol. These loci were polymorphic in M. demerarae and Micoureus paraguayanus populations from the Atlantic Forest in Brazil with the number of alleles ranging from two to 23. Those eight loci plus another five already described for M. paraguayanus will allow for the evaluation of genetic diversity of populations from the ‘Rio Doce’ Park, one of the last Atlantic Forest fragments in Minas Gerais state, Brazil.