Geographic distribution of hantaviruses associated with neotomine and sigmodontine rodents, Mexico.

To increase our knowledge of the geographic distribution of hantaviruses associated with neotomine or sigmodontine rodents in Mexico, we tested 876 cricetid rodents captured in 18 Mexican states (representing at least 44 species in the subfamily Neotominae and 10 species in the subfamily Sigmodontinae) for anti-hantavirus IgG. We found antibodies against hantavirus in 35 (4.0%) rodents. Nucleotide sequence data from 5 antibody-positive rodents indicated that Sin Nombre virus (the major cause of hantavirus pulmonary syndrome [HPS] in the United States) is enzootic in the Mexican states of Nuevo León, San Luis Potosí, Tamaulipas, and Veracruz. However, HPS has not been reported from these states, which suggests that in northeastern Mexico, HPS has been confused with other rapidly progressive, life-threatening respiratory diseases. Analyses of nucleotide sequence data from 19 other antibody-positive rodents indicated that El Moro Canyon virus and Limestone Canyon virus are geographically widely distributed in Mexico.

Reevaluation of the phylogenetic relationships among Neotomini rodents (Hodomys, Neotoma, and Xenomys) and comments on the woodrat classification.

The woodrats or packrats of the genus Neotoma have been the subject of a wide array of research including paleoecology, physiology, morphological evolution, systematics, speciation, and hybridization. In recent years, much work has been done to elucidate evolutionary relationships within and between closely related species of the genus; in particular the addition of newly collected specimens from critical geographic regions has provided new opportunities for taxonomic assessment. Given these new data and their potential, parsimony (PARS), maximum likelihood (ML), and Bayesian inference (BI) analyses were conducted on DNA sequences obtained from nine individual genes (four mitochondrial loci: 12S, 16S, CoII, and Cytb; five nuclear loci: AdhI2, BfibI7, En2, Mlr, and Myh6) to estimate the phylogenetic relationships among 23 species of Neotoma. Results of these analyses depicted a wide array of phylogenetic relationships among taxa; with substantial nodal support recovered in both the ML and PARS analyses at some mid-level and terminal positions. Several individual genes, particularly 12S, AdhI2, BfibI7, CoII, and Cytb, provided support at several basal positions; however, phylogenetic resolution was limited in the other genes. A final BI analysis where the nine genes were concatenated into a single data set produced several supported clades that corresponded to previously recognized species groups (floridana, micropus, mexicana, and lepida) and the subgenus Homodontomys. Levels of genetic divergence for within-species comparisons (estimated from the Cytb data set) ranged from 0.88% (N. magister) to 6.82% (N. fuscipes); for between sister species comparisons ranged from 4.68% (N. devia and N. lepida) to 12.70% (N. angustapalata and N. nelsoni); and for members within closely related clades ranged from 8.70% (N. bryanti and N. lepida) to 12.57% (N. goldmani and N. magister). Evaluations of generic, subgeneric, and species group boundaries were explored using phylogenetic principles on the DNA sequence data presented herein, as well as morphological findings from previous studies. Results obtained suggest that the most conservative taxonomic interpretation involves the abandonment of subgeneric delineations and relies on the recognition of eight species groups (cinerea, floridana, fuscipes, lepida, mexicana, micropus, phenax, and stephensi) as the backbone of the woodrat classification.

Las ratas cambalacheras del género Neotoma han sido estudiadas en varios tipos de investigaciones incluyendo paleoecología, fisiología, evolución morfológica, sistemática, especiación e hibridación. Recientemente, se han realizado numerosos estudios para elucidar las relaciones evolutivas dentro del género y entre especies cercanamente relacionadas al mismo; en particular la inclusión de nuevos especímenes provenientes de regiones geográficas críticas han brindado nuevas oportunidades para evaluaciones taxonómicas. A partir de estos nuevos datos se realizaron análisis de parsimonia (PARS), Máxima Verosimilitud (MV), e Inferencia Bayesiana (IB) en secuencias de ADN provenientes de nueve genes individuales (cuatro loci mitocondriales: 12S, 16S, CoII, y Cytb; cinco loci nucleares: Adh-I2, Bfib-I7, En2, Mlr, and Myh6) para determinar la relación filogenética de 23 especies de Neotoma. Los resultados de estos análisis presentan una amplia gama de relaciones filogenéticas entre taxa con un soporte nodal importante en los análisis de MV y PARS en algunas posiciones terminales de nivel medio. Varios genes individuales, en particular 12S, Adh-I2, Bfib-I7, CoII, and Cytb, ofrecieron soporte en varias posiciones basales; sin embargo, la resolución filogenética fue reducida en los demás genes. El último análisis de IB, en donde nueve genes se concatenaron en un solo conjunto de datos, produjo soporte en varios clados que correspondieron a especies de grupos previamente reconocidos (floridana, micropus, mexicana, y lepida) y el sub-género Homodontomys. Los niveles de divergencia genética para comparaciones intraespecíficas fluctuaron entre 0.88% (N. magister) y 6.82% (N. fuscipes); para especies hermanas (4.68%­N. devia y N. lepida hasta 12.70%­N. angustapalata y N. nelsoni); y para los miembros de clados cercanos (8.70%­N. bryanti y N. lepida hasta 12.57%­N. goldmani y N. magister). Las evaluaciones de los limites genéricos, subgenéricos y de grupos de especies fueron explorados usando principios filogenéticos en las secuencias de ADN de este trabajo, y también se basaron en las conclusiones morfológicas de estudios previos. Los resultados obtenidos sugieren que la interpretación taxonómica más conservadora incluye el abandono de las delineaciones subgenéricas y se depende en el reconocimiento de ocho grupos de especies (cinerea, floridana, fuscipes, lepida, mexicana, micropus, phenax, y stephensi) como el pilar central de la clasificación de las ratas cambalacheras.

Transmission of Guanarito and Pirital viruses among wild rodents, Venezuela.

Samples from rodents captured on a farm in Venezuela in February 1997 were tested for arenavirus, antibody against Guanarito virus (GTOV), and antibody against Pirital virus (PIRV). Thirty-one (48.4%) of 64 short-tailed cane mice (Zygodontomys brevicauda) were infected with GTOV, 1 Alston's cotton rat (Sigmodon alstoni) was infected with GTOV, and 36 (64.3%) of 56 other Alston's cotton rats were infected with PIRV. The results of analyses of field and laboratory data suggested that horizontal transmission is the dominant mode of GTOV transmission in Z. brevicauda mice and that vertical transmission is an important mode of PIRV transmission in S. alstoni rats. The results also suggested that bodily secretions and excretions from most GTOV-infected short-tailed cane mice and most PIRV-infected Alston's cotton rats may transmit the viruses to humans.

Catacamas virus, a hantaviral species naturally associated with Oryzomys couesi (Coues' oryzomys) in Honduras.

The primary objective of this study was to extend our knowledge of the geographical distribution, genetic diversity, and natural host associations of the hantaviruses indigenous to North America. Antibody to a hantavirus was found in 5 (20.8%) of 24 Coues' oryzomys (Oryzomys couesi) and none of 41 other rodents captured near the town of Catacamas in eastern Honduras, and a hantavirus was isolated from one of the antibody-positive Coues' oryzomys. Analyses of nucleotide and amino acid sequence data indicated that the viral isolate is a strain of a novel hantaviral species (proposed species name "Catacamas virus") that is phylogenetically most closely related to Bayou virus, a hantaviral species that is principally associated with Oryzomys palustris (marsh oryzomys) in the southeastern United States. Catacamas virus is the first evidence for the occurrence of a hantaviral species in Honduras and the first evidence that a hantaviral species is naturally associated with an Oryzomys species other than O. palustris.

Maporal virus, a hantavirus associated with the fulvous pygmy rice rat (Oligoryzomys fulvescens) in western Venezuela.

Oryzomine rodents in the southeastern United States, Panama, and southern South America are natural hosts of 6 of the 13 viruses known to cause hantavirus pulmonary syndrome. The purpose of this study was to extend our knowledge of the geographical distribution and genetic diversity of the hantaviruses associated with oryzomine rodents in South America. An infectious hantavirus was isolated from two fulvous pygmy rice rats captured in western Venezuela. Analyses of complete nucleocapsid protein and glycoprotein precursor sequences indicated that the isolates are strains of a novel hantavirus (proposed name "Maporal") which is phylogenetically most closely related to the viruses known to cause hantavirus pulmonary syndrome in southern South America.

Experimental infection of the Sigmodon alstoni cotton rat with Caño Delgadito virus, a South American hantavirus.

Forty-eight Sigmodon alstoni (Alston's cotton rat) were inoculated with Caño Delgadito (CDG) virus to extend our knowledge and understanding of the natural host relationships of the hantaviruses indigenous to the Americas. Infectious CDG virus was recovered from oropharyngeal secretions, urine, or solid tissues of nine of 12 animals killed on day 9 post-inoculation (PI), 14 of 24 animals killed on day 18 or 27 PI, and none of 12 animals killed on day 54 PI. In addition, virus-specific RNA was detected in the kidneys of six of the 12 animals killed on day 54 PI, and adult cotton rats inoculated with the kidneys of four animals killed on day 54 PI developed antibody to CDG virus. Collectively, the results indicate that CDG virus can establish lengthy (perhaps lifelong) infections in Alston's cotton rat and thus support the concept that S. alstoni is the principal host of CDG virus.

Epizootiology of arenaviral infections in the white-throated woodrat (Muridae: Sigmodontinae) and other woodrats in Arizona.

The purpose of this study was to extend and refine our knowledge of the geographical distribution and natural host relationships of the arenaviruses associated with woodrats indigenous to Arizona. Antibody to a Tacaribe serocomplex virus was found in 112 of 1,250 white-throated woodrats, five of 208 Mexican woodrats, one of 114 Stephen's woodrats, and none of 862 other rodents captured at 51 sites in 10 counties in Arizona. Of the 112 antibody-positive white-throated woodrats, 109 (97.3%) were captured within extensive, dense patches of prickly pear cactus and cane cholla in three counties in mid-central Arizona. Analysis of the serological and zoographical data suggested that white-throated woodrats usually become infected early in life and that the distribution of antibody-positive white-throated woodrats in Arizona is not linked to a specific biome.

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.

Duration of Catarina virus infection in the southern plains woodrat (Neotoma micropus).

Four adult male, 6 sub-adult, and 7 newborn southern plains woodrats (Neotoma micropus) each were inoculated subcutaneously with 3.1 log(10) median cell culture infectious doses (CCID(50)) of Catarina virus strain AV A0400135 (virus family Arenaviridae). The inoculated animals and the mothers of the newborn animals all became infected and remained asymptomatic. The infections in the adult male woodrats and in the mother woodrats were transient, the infections in 2 (33.3%) of the sub-adult woodrats persisted through month 4 post-inoculation, and 6 (85.7%) of the newborn woodrats were viruric through month 5 post-inoculation. Collectively these findings indicate that the duration of infection in the southern plains woodrat is dependent upon age at exposure to Catarina virus. The results of this study also indicate that chronically infected woodrats persistently shed Catarina virus into the environment.

Natural host relationships of hantaviruses native to western Venezuela.

Strains of Caño Delgadito virus (CADV) and Maporal virus (MAPV) were isolated from 25 (8.9%) of the 280 rodents captured on farms in 1997 in western Venezuela. The results of analyses of laboratory and zoographic data indicated that Alston's cotton rat (Sigmodon alstoni) is the principal host of CADV, horizontal virus transmission is the dominant mode of CADV transmission in Alston's cotton rat in nature, a pygmy rice rat (Oligoryzomys sp.) is the principal host of MAPV, and the natural host relationships of CADV and MAPV are highly specific.

Pathology of Black Creek Canal virus infection in juvenile hispid cotton rats (Sigmodon hispidus).

The purpose of this study was to assess the effect of inoculum dose on the pathogenesis of Black Creek Canal virus (BCCV) infection in the hispid cotton rat (Sigmodon hispidus), the principal host of BCCV. No sign of illness was observed in any of the 52 juvenile hispid cotton rats inoculated with 3.1, 1.1, -0.9, or -2.9 log(10) median infectious doses(VeroE6) (ID(50-VeroE6)) of BCCV and euthanized on day 9, 18, 27, or 54 postinoculation (PI). Analysis of virus assay and serological data indicated that inoculum dose could significantly affect the pathogenesis of BCCV infection in juvenile hispid cotton rats. For example, the six animals inoculated with 3.1 or 1.1 log(10) ID(50-VeroE6) and euthanized on day 54 PI were virus positive and antibody positive, whereas the six animals inoculated with -0.9 or -2.9 log(10) ID(50-VeroE6) and euthanized on day 54 PI were virus positive but antibody negative. Microscopic examination of tissues from the animals inoculated with 3.1 or 1.1 log(10) ID(50-VeroE6) revealed diffuse, subacute pneumonitis in the lungs of all the animals euthanized on day 18 PI or thereafter, and indicated that the severity of pneumonitis was dependent upon inoculum dose as well as duration of infection (i.e., amount of time elapsed since inoculation).

Antibodies to Tacaribe serocomplex viruses (family Arenaviridae, genus Arenavirus) in cricetid rodents from New Mexico, Texas, and Mexico.

Blood samples from 4893 cricetid rodents were tested for antibody (immunoglobulin G) to Whitewater Arroyo virus and Amaparí virus to extend our knowledge of the natural host range and geographical distribution of Tacaribe serocomplex viruses in North America. Antibodies to arenaviruses were found in northern pygmy mice (Baiomys taylori), woodrats (Neotoma spp.), northern grasshopper mice (Onychomys leucogaster), oryzomys (Oryzomys spp.), deermice (Megadontomys nelsoni and Peromyscus spp.), harvest mice (Reithrodontomys spp.), and cotton rats (Sigmodon spp.) captured in New Mexico, Texas, or Mexico. Comparison of endpoint antibody titers to Whitewater Arroyo virus and Amaparí virus in individual blood samples indicated that the Tacaribe complex viruses enzootic in Texas and Mexico are antigenically diverse.

Molecular Evidence For High Levels of Intrapopulation Genetic Diversity in Woodrats (Neotoma Micropus).

Nucleotide sequences from the mitochondrial control region and genotypes from 5 nuclear microsatellite loci were used to examine genetic structure and infer recent (within approximately the last 3,000 years) evolutionary history of a population (549 individuals) of the southern plains woodrat (Neotoma micropus). Observed heterozygosity values ranged from 0.61 to 0.89 across microsatellite loci and systematically were lower than expected heterozygosity values (0.66-0.95). Probability of unique identity using microsatellite data was high (1 individual in 66,005,424). Fifty-three mitochondrial haplotypes were obtained from 150 individuals. F(ST) values estimated from sequence and microsatellite data were 0.061 and 0.011, respectively, and the R(ST) for microsatellite data was 0.007. Within-group genetic variation ranged from 93.90% to 99.99% depending on whether sequence or microsatellite data were examined. Analyses of microsatellite data suggested that all sampled individuals belonged to a single population, albeit genetically diverse. However, combined data analyses suggested the presence of low levels of substructure attributable to maternal lineages within the population. Low nucleotide-diversity values (0.007-0.010) in addition to high haplotype-diversity values (0.915-0.933) indicate a high number of closely related haplotypes, and suggest that this population may have undergone a recent expansion. However, Fu's F(S) statistic did not fully support this finding, because it did not reveal a significant excess of recent mutations. A phylogenetic approach using the haplotype sequence data and a combined set including both haplotype and genotype data was used to test for evolutionary patterns and history.

GENETIC DIVERSITY WITHIN THE SOUTHERN PLAINS WOODRAT (NEOTOMA MICROPUS) IN SOUTHERN TEXAS.

Genetic diversity within a population of the southern plains woodrat was examined using DNA sequences (967 base pairs [bp]) obtained from the control or d-loop region of the mitochondrial genome. One hundred fourteen individuals from 10 collection sites were assigned to 42 haplotypes. Haplotype diversity values were moderate to high (0.974 overall and ranged from 0.524 to 0.964 across collecting sites), whereas nucleotide diversity values were low (0.008 overall and ranged from 0.001 to 0.010 across sites), indicating that this population possesses a high number of closely related haplotypes. Seventy-nine percent of the genetic variability was partitioned within groups that corresponded to the collecting sites. In addition, 13 samples from Texas, New Mexico, and Mexico were included as references for evaluating the evolutionary history of haplotypes. Nested clade analysis revealed that restricted gene flow with isolation by distance in conjunction with contiguous range expansion was responsible for the observed pattern of genetic diversity. A test of neutrality supported the diagnosis of restricted gene flow, but failed to support contiguous range expansion due solely to population growth. Examination of the spatial distribution of the haplotypes indicated that most haplotypes were restricted to a single collecting site; however, a small number of haplotypes were found at 2 or more sites. A phylogenetic analysis indicated that some haplotypes (28.6%) were restricted to the study area whereas the remaining haplotypes occupied a broader geographic region.

Bear Canyon virus: an arenavirus naturally associated with the California mouse (Peromyscus californicus).

Thirty-four rodents captured in southern California were studied to increase our knowledge of the arenaviruses indigenous to the western United States. An infectious arenavirus was isolated from 5 of 27 California mice but none of the 7 other rodents. Analyses of viral nucleocapsid protein gene sequence data indicated that the isolates from the California mice are strains of a novel Tacaribe serocomplex virus (proposed name "Bear Canyon") that is phylogenetically most closely related to Whitewater Arroyo and Tamiami viruses, the only other Tacaribe serocomplex viruses known to occur in North America. The discovery of Bear Canyon virus is the first unequivocal evidence that the virus family Arenaviridae is naturally associated with the rodent genus Peromyscus and that a Tacaribe serocomplex virus occurs in California.