Multilocus Characterization of a Woodrat (Genus Neotoma) Hybrid Zone.

In order to investigate hybridization between 2 species of woodrats, Neotoma floridana and Neotoma micropus, 103 specimens were collected, in March of 1988, from a known area of sympatry, and compared with reference collections from areas of allopatry. Ten genetic markers, consisting of 7 microsatellite loci, 1 mitochondrial gene (cytochrome-b [Cytb]), and 2 nuclear introns (intron 2 of the vertebrate alcohol dehydrogenase gene [Adh1-I2] and intron 7 of the beta-fibrinogen gene [Fgb-I7]) were used to develop a composite genotype for each individual and for detection of hybridization. Six individuals were identified as pure parental N. micropus, 96 as hybrids, and 1 as pure parental N. floridana Hybrids were formed primarily through matings between complex genotypes, resulting in a high prevalence of individuals classified as backcrosses. The ratio of hybrid classes, population substructure, and presence of significant linkage disequilibrium within the zone of contact could not reject either the hybrid superiority or hybrid equilibrium model as responsible for maintenance of this hybrid zone. The collection date of this dataset (1988) provided not only a point in time assessment of the hybrid zone but also provided opportunities for future comparisons of temporal datasets with the purpose of examining hybrid zone characteristics over multiple generations.

Trypanosoma cruzi in a Mexican Free-Tailed Bat ( Tadarida brasiliensis) in Oklahoma, USA.

Trypanosoma cruzi is a vector-borne protozoan parasite that infects seven million individuals in Central and South America and is the etiologic agent of Chagas disease. There are increasing reports of endemic transmission within the southern US. Trypanosoma cruzi occurs in wild raccoons and dogs in Oklahoma, but its endemicity in the state is poorly studied. We suspected Mexican free-tailed bats ( Tadarida brasiliensis) contributed to the endemicity of T. cruzi in Oklahoma due to their annual migration from Central America to their North American maternity roosts. During the summer of 2017, we sampled 361 Mexican free-tailed bats for T. cruzi at three maternity roosts in Oklahoma. We collected wing tissues, extracted T. cruzi DNA, amplified target DNA by PCR using the primers TCZ1/TCZ2, and observed amplification by gel electrophoresis. One juvenile Mexican free-tailed bat was positive for T. cruzi resulting in a 0.27% prevalence in the 361 sampled bats. Our finding of a wild bat naturally infected with T. cruzi in Oklahoma provided insight on the endemicity of T. cruzi in underrepresented endemic areas. The positive sample was sequenced, confirmed as T. cruzi, and uploaded to GenBank (no. MG869732). Future research will focus on monitoring T. cruzi prevalence in wild bats and insect vectors to better understand the enzootic emergence of this neglected tropical parasite.

Ecology of Catarina virus (family Arenaviridae) in southern Texas, 2001-2004.

A total of 3941 rodents were captured during a 46-month prospective (mark-recapture) study on the ecology of Catarina virus in southern Texas. Antibody reactive against Catarina virus was found in 73 (11.9%) of 611 southern plains woodrats (Neotoma micropus) and none of 3330 other rodents; strains of Catarina virus were isolated from 6 antibody-negative and 9 antibody-positive southern plains woodrats; and the infections in at least 3 southern plains woodrats were chronic. These results affirm the notion that the southern plains woodrat is the principal host of Catarina virus and suggest that Catarina virus infection is highly specific to N. micropus.

Diversity and phylogenetic relationships among the North American Tacaribe serocomplex viruses (family Arenaviridae).

The purpose of this study was to extend our knowledge of the genetic diversity and phylogenetic relationships among the North American Tacaribe serocomplex viruses. Analyses of glycoprotein precursor gene sequence data separated the North American arenaviruses into 7 major phylogenetic groups. The results of analyses of Z gene and nucleocapsid protein gene sequence data were not remarkably different from the glycoprotein precursor gene tree. In contrast, the tree generated from RNA-dependent RNA polymerase gene sequences differed from the glycoprotein precursor gene tree with regard to phylogenetic relationships among the viruses associated with woodrats captured in the western United States, Texas, or northern Mexico. Further analyses of the polymerase gene sequence data set suggested that the difference in topology was a consequence of incongruence among the gene tree data sets or chance rather than genetic reassortment or recombination between arenaviruses.

Diversity among tacaribe serocomplex viruses (family Arenaviridae) naturally associated with the white-throated woodrat (Neotoma albigula) in the southwestern United States.

Bayesian analyses of glycoprotein precursor and nucleocapsid protein gene sequences indicated that arenaviruses naturally associated with white-throated woodrats in central Arizona are phylogenetically closely related to the Whitewater Arroyo virus prototype strain AV 9310135, which originally was isolated from a white-throated woodrat captured in northwestern New Mexico. Pairwise comparisons of glycoprotein precursor and nucleocapsid protein amino acid sequences revealed extensive diversity among arenaviruses isolated from white-throated woodrats captured in different counties in central Arizona and extensive diversity between these viruses and Whitewater Arroyo virus strain AV 9310135. It was concluded that the viruses isolated from the white-throated woodrats captured in Arizona represent 2 novel species (Big Brushy Tank virus and Tonto Creek virus) and that these species should be included with Whitewater Arroyo virus in a species complex within the Tacaribe serocomplex (family Arenaviridae, genus Arenavirus).

GENETIC VARIATION IN MULTILOCUS MICROSATELLITE GENOTYPES IN TWO SPECIES OF WOODRATS (NEOTOMA MACROTIS AND N. FUSCIPES) FROM CALIFORNIA.

Five microsatellite loci were used to develop multilocus genotypes for Neotoma macrotis (n = 128) and N. fuscipes (n = 29). Several statistical analyses were used to estimate genetic structure, levels of genetic variability, and degree of relatedness within groups of these 2 species. Samples of N. macrotis represented 2 groups and 4 population clusters throughout southern California. Samples of N. fuscipes represented 2 regions in northern and southern California. Genetic structure was detected among samples of N. macrotis and N. fuscipes at a regional level. Both species displayed moderate to high genetic diversity in terms of mean expected heterozygosity (0.939 and 0.804 for N. macrotis and N. fuscipes, respectively) and mean polymorphic information content (0.930 and 0.761 for N. macrotis and N. fuscipes, respectively). Mean relatedness values within regions and populations of N. macrotis indicated 4th-order levels of relatedness within groups (e.g., distant-cousin relationships). Mean relatedness values within regions of N. fuscipes indicated 2nd-order (e.g., half-sibling) relationships within the northern region and 3rd-order (e.g., cousin) relationships in the southern region. One locus in particular (Nma04) was determined to be diagnostic in distinguishing between these 2 species.

Experimental Everglades virus infection of cotton rats (Sigmodon hispidus).

Everglades virus (EVEV), an alphavirus in the Venezuelan equine encephalitis (VEE) serocomplex, circulates among rodents and vector mosquitoes and infects humans, causing a febrile disease sometimes accompanied by neurologic manifestations. EVEV circulates near metropolitan Miami, which indicates the potential for substantial human disease, should outbreaks arise. We characterized EVEV infection of cotton rats in South Florida, USA, to validate their role in enzootic transmission. To evaluate whether the viremia induced in cotton rat populations regulates EVEV distribution, we also infected rats from a non-EVEV-endemic area. Viremia levels developed in rats from both localities that exceeded the threshold for infection of the vector. Most animals survived infection with no signs of illness, despite virus invasion of the brain and the development of mild encephalitis. Understanding the mechanisms by which EVEV-infected cotton rats resist clinical disease may be useful in developing VEE therapeutics for equines and humans.