E. Gus Cothran

INFLUENCES OF GENETIC VARIABILITY AND MATERNAL FACTORS ON FETAL GROWTH IN WHITE-TAILED DEER

A number of quantitative traits influence secondary productivity and are associated with genetic variability. High heterozygosity has been positively correlated with aggressiveness and exploratory behavior (Garten, 1976, 1977), social dominance (Baker and Fox, 1978), body size (Koehn et al., 1973; Boyer, 1974; Garten, 1976), growth rate (Singh and Zouros, 1978), production of offspring (Smith et al., 1975; Johns et al., 1977) and developmental stability (Mitton, 1978; Soule, 1979). In domestic animals, much of the variation in secondary productivity (i.e., somatic growth) has been explained by the effects of inbreeding depression or heterosis (Falconer, 1960). Reproductive capacity and/or physiological efficiency are apparently the characteristics most sensitive to inbreeding depression (Falconer, 1960 p. 258). Secondary productivity and the heterozygosity of individuals may be influenced by the breeding history of the population. Differences in gene frequency over relatively short distances indicate population subdivision and some degree of site tenacity for individual animals, since dispersal between subdivisions results in genetic homogeneity in the absence of large differences in selective pressures (Crow and Kimura, 1970). The combination of site tenacity and small population size increases the likelihood of inbreeding. Dispersal between the subdivisions will increase the heterozygosity of resulting offspring and eliminate the direct effects of inbreeding (i.e., apparent heterosis). In-

Microsatellites in Portuguese autochthonous horse breeds: usefulness for parentage testing

The use of DNA technology for parentage testing is increasing every day. Most laboratories have improved their exclusion probabilities (PE) by the addition of DNA microsatellite loci to standard blood-typing results. The efficacy of each locus depends on the number of alleles detected and their frequencies in the breeds tested. Here we analyzed the usefulness of six microsatellites for routine parentage testing in three Portuguese autochthonous horse breeds: Lusitano, Sorraia and Garrano. The DNA loci analyzed - ASB2, HMS3, HMS7, HTG4, HTG10, and VHL20 - were chosen based upon the polymorphism detected in other breeds with Iberian horse influence. The estimated probability of exclusion of wrongly named parents (PE) was high, with values ranging from 88.5% to 99.6%.

Variation in the mitochondrial control region sequence between the two maternal lines of the Sorraia horse breed

The mitochondrial control region sequence was compared between the two surviving matrilineal lineages of the Sorraia horse. Two haplotypes were found, showing three differences between them. The reduced number of surviving maternal lineages emphasizes the importance of establishing a conservation plan for this endangered breed.

Genetic variability in white-tailed deer.

Genetic variability at 36 loci was studied in white-tailed deer (Odocoileus virginianus) populations, on the Savannah River Plant (SRP) in South Carolina. Mean multilocus heterozygosity ([Hmacr ]), percentage of polymorphic loci (P) and average number of alleles per locus (A) for white-tailed deer were calculated and compared with values for white-tailed deer from several locations (SRP deer: [Hmacr ] = 9·9 per cent, P0·05 = 30.6 per cent, A = 1·89; white-tailed deer overall: [Hmacr ] = 10·4 per cent, P0·05 = 32.3 per cent, A = 1·94). Frequency distributions for single locus heterozygosity values (h) and the number of alleles per locus for white-tailed deer were found to be significantly different from those of mammals in general. Analysis of single-locus data based on quaternary structure and functional groups of proteins failed to demonstrate expected differences as predicted from the literature. White-tailed deer have a high level of heterozygosity, but they do not exhibit many of the life history and environmental characteristics associated with high heterozygosity in other animals.

Mitochondrial DNA D-loop sequence variation among 5 maternal lines of the Zemaitukai horse breed

Genetic variation in Zemaitukai horses was investigated using mitochondrial DNA (mtDNA) sequencing. The study was performed on 421 bp of the mitochondrial DNA control region, which is known to be more variable than other sections of the mitochondrial genome. Samples from each of the remaining maternal family lines of Zemaitukai horses and three random samples for other Lithuanian (Lithuanian Heavy Draught, Zemaitukai large type) and ten European horse breeds were sequenced. Five distinct haplotypes were obtained for the five Zemaitukai maternal families supporting the pedigree data. The minimal difference between two different sequence haplotypes was 6 and the maximal 11 nucleotides in Zemaitukai horse breed. A total of 20 nucleotide differences compared to the reference sequence were found in Lithuanian horse breeds. Genetic cluster analysis did not shown any clear pattern of relationship among breeds of different type.

Genetic Analysis of Three Lithuanian Native Horse Breeds

This study was the first to analyze the polymorphic characteristics of a wide range of genetic markers in three Lithuanian horse breeds. Genetic variation at seven red blood cell alloantigen (blood group) loci, 10 biochemical genetic loci and 16 microsatellites was examinated in 31 Zemaitukai, 30 large-type Zemaitukai and 24 Lithuanian Heavy Draught horse and compared to that of other domestic horse breeds. Genetic variability of the Zemaitukai breeds based upon blood groups and biochemical loci was higher than average for domestic horse breeds. For microsatellite loci, the variability of the large-type Zemaitukai and the Heavy Draught breeds also was high compared to the mean variation for domestic breeds; however, for the Zemaitukai breed, variation levels were very near the average for horses.

Genetics of the Pantaneiro horse of the Pantanal region of Brazil

Genetic variation at seven red blood cell alloantigen, blood group loci and 10 biochemical genetic loci was examined in 102 Pantaneiro horses from the Pantanal region of Brazil and compared to that of other domestic horse breeds. Measures of both individual genic variation and populational genetic diversity within the Pantaneiro were near the average for domestic horse breeds. From the standpoint of genetic conservation there is no immediate concern for loss of variation within the Pantaneiro. Analysis of genetic relationship showed that the Pantaneiro was clearly of Iberian Peninsula descent and that it was most closely related to other Brazilian breeds.

Maternal Lineages in Native Canadian Equine Populations and Their Relationship to the Nordic and Mountain and Moorland Pony Breeds

A 378-bp section of the mitochondrial displacement loop was used to estimate genetic diversity in the native Canadian equine populations. The inclusion of 10 Mountain and Moorland, 3 Nordic pony breeds, 2 feral populations, and 5 horse breeds were also investigated as they may have influenced the development (or rejuvenation) of the native Canadian populations. A total of 281 samples were sequenced, which produced 75 haplotypes derived from 54 informative sites. On further investigation, 36 of these 75 haplotypes were found to be previously unreported. Overall, total diversity was lowest in the feral Sable Island population with a haplotype diversity (0.27 ± 0.12), nucleotide diversity (0.0007 ± 0.0004), and pairwise difference of 0.286 ± 0.317. This is not surprising due to the geographic isolation of this population. Haplotype diversity was highest (1.00 ± 0.13) in the New Forest population, pairwise difference was highest (8.061 ± 4.028) in the Icelandic breed, whereas nucleotide diversity was highest in the Exmoor breed (0.0209 ± 0.0025). Within the Canadian populations, haplotype diversity was highest in the Newfoundland pony (0.96 ± 0.08), whereas pairwise difference and nucleotide diversity was highest in the Canadian horse (7.090 ± 3.581 and 0.0188 ± 0.0042, respectively). Three different estimates of genetic distances were used to examine the phylogenetic relationships amongst these populations. All 3 estimates produced similar topologies. In general, the native Canadian populations were highly represented in the D clade, with particular emphasis in the D1 and D2 clades. This is an important factor when considering the phylogenetic conservation of these Canadian equine populations.

Characterization of expressed sequence tags generated from skin cDNA clones of Equus caballus by single pass sequencing.

A cDNA library was built using RNA extracted from the skin tissue of an adult horse. The library was primed with oligo (dT) and sequences were directionally inserted in order to produce an expression library. The library has 5.8X 105 plaque forming units with 99.6% recombinant phage. The average insert size is 1.3 Kbp. Three hundred and thirteen expressed sequence tags (ESTs) were generated from sequencing of the 5 prime end of randomly selected skin cDNA clones. The ESTs were sequenced on an ABI 377 using Big-Dye chemistry. A similarity search was performed on each EST using the NCBI non-redundant protein database and 206 ESTs were putatively identified. Twenty six percent of the identified ESTs were redundant. The ESTs were categorized by function. The most frequently identified functional class was translational proteins.A cDNA library was built using RNA extracted from the skin tissue of an adult horse. The library was primed with oligo (dT) and sequences were directionally inserted in order to produce an expression library. The library has 5.8X 105 plaque forming units with 99.6% recombinant phage. The average insert size is 1.3 Kbp. Three hundred and thirteen expressed sequence tags (ESTs) were generated from sequencing of the 5 prime end of randomly selected skin cDNA clones. The ESTs were sequenced on an ABI 377 using Big-Dye chemistry. A similarity search was performed on each EST using the NCBI non-redundant protein database and 206 ESTs were putatively identified. Twenty six percent of the identified ESTs were redundant. The ESTs were categorized by function. The most frequently identified functional class was translational proteins.

Genetic diversity and origin of the feral horses in Theodore Roosevelt National Park

Feral horses in Theodore Roosevelt National Park (TRNP) represent an iconic era of the North Dakota Badlands. Their uncertain history raises management questions regarding origins, genetic diversity, and long-term genetic viability. Hair samples with follicles were collected from 196 horses in the Park and used to sequence the control region of mitochondrial DNA (mtDNA) and to profile 12 autosomal short tandem repeat (STR) markers. Three mtDNA haplotypes found in the TRNP horses belonged to haplogroups L and B. The control region variation was low with haplotype diversity of 0.5271, nucleotide diversity of 0.0077 and mean pairwise difference of 2.93. We sequenced one mitochondrial genome from each haplotype determined by the control region. Two complete mtDNA sequences of haplogroup L were closely related to the mtDNA of American Paint horse. The TRNP haplotype B did not have close matches in GenBank. The phylogenetic test placed this sequence in a group consisting of two horses from China, one from Yakutia, and one from Italy raising a possibility of historical transportation of horses from Siberia and East Asia to North America. Autosomal STR loci were polymorphic and indicated that the TRNP horses were distinctly different from 48 major horse breeds. Heterozygosity, mean number of alleles, and other measures of diversity indicated that TRNP herd diversity was below that observed for most other feral herds and domestic breeds. Both mtDNA and STRs demonstrated that the existing genetic data sets of horses are insufficient to determine the exact origins of the TRNP horses. However, measures of nuclear and mitochondrial diversity have elucidated management needs. It is recommended that new genetic stock be introduced and that adaptive management principles are employed to ensure that unique mitochondrial lineages are preserved and genetic diversity is increased and maintained over time.