Deep Sequencing to Reveal Phylo-Geographic Relationships of Juquitiba Virus in Paraguay.

Several hantaviruses result in zoonotic infections of significant public health concern, causing hemorrhagic fever with renal syndrome (HFRS) or hantavirus cardiopulmonary syndrome (HCPS) in the Old and New World, respectively. Given a 35% case fatality rate, disease-causing New World hantaviruses require a greater understanding of their biology, genetic diversity, and geographical distribution. Juquitiba hantaviruses have been identified in Oligoryzomys nigripes in Brazil, Paraguay, and Uruguay. Brazil has reported the most HCPS cases associated with this virus. We used a multiplexed, amplicon-based PCR strategy to screen and deep-sequence the virus harbored within lung tissues collected from Oligoryzomys species during rodent field collections in southern (Itapúa) and western (Boquerón) Paraguay. No Juquitiba-like hantaviruses were identified in Boquerón. Herein, we report the full-length S and M segments of the Juquitiba hantaviruses identified in Paraguay from O. nigripes. We also report the phylogenetic relationships of the Juquitiba hantaviruses in rodents collected from Itapúa with those previously collected in Canindeyú. We showed, using the TN93 nucleotide substitution model, the coalescent (constant-size) population tree model, and Bayesian inference implemented in the Bayesian evolutionary analysis by sampling trees (BEAST) framework, that the Juquitiba virus lineage in Itapúa is distinct from that in Canindeyú. Our spatiotemporal analysis showed significantly different time to the most recent ancestor (TMRA) estimates between the M and S segments, but a common geographic origin. Our estimates suggest the additional geographic diversity of the Juquitiba virus within the Interior Atlantic Forest and highlight the need for more extensive sampling across this biome.

Impact of Predator Exclusion and Habitat on Seroprevalence of New World Orthohantavirus Harbored by Two Sympatric Rodents within the Interior Atlantic Forest.

Understanding how perturbations to trophic interactions influence virus-host dynamics is essential in the face of ongoing biodiversity loss and the continued emergence of RNA viruses and their associated zoonoses. Herein, we investigated the role of predator exclusion on rodent communities and the seroprevalence of hantaviruses within the Reserva Natural del Bosque Mbaracayú (RNBM), which is a protected area of the Interior Atlantic Forest (IAF). In the IAF, two sympatric rodent reservoirs, Akodon montensis and Oligoryzomys nigripes, harbor Jaborá and Juquitiba hantavirus (JABV, JUQV), respectively. In this study, we employed two complementary methods for predator exclusion: comprehensive fencing and trapping/removal. The goal of exclusion was to preclude the influence of predation on small mammals on the sampling grids and thereby potentially reduce rodent mortality. Following baseline sampling on three grid pairs with different habitats, we closed the grids and began predator removal. By sampling three habitat types, we controlled for habitat-specific effects, which is important for hantavirus-reservoir dynamics in neotropical ecosystems. Our six-month predator exclusion experiment revealed that the exclusion of terrestrial mammalian predators had little influence on the rodent community or the population dynamics of A. montensis and O. nigripes. Instead, fluctuations in species diversity and species abundances were influenced by sampling session and forest degradation. These results suggest that seasonality and landscape composition play dominant roles in the prevalence of hantaviruses in rodent reservoirs in the IAF ecosystem.

Common Themes in Zoonotic Spillover and Disease Emergence: Lessons Learned from Bat- and Rodent-Borne RNA Viruses.

Rodents (order Rodentia), followed by bats (order Chiroptera), comprise the largest percentage of living mammals on earth. Thus, it is not surprising that these two orders account for many of the reservoirs of the zoonotic RNA viruses discovered to date. The spillover of these viruses from wildlife to human do not typically result in pandemics but rather geographically confined outbreaks of human infection and disease. While limited geographically, these viruses cause thousands of cases of human disease each year. In this review, we focus on three questions regarding zoonotic viruses that originate in bats and rodents. First, what biological strategies have evolved that allow RNA viruses to reside in bats and rodents? Second, what are the environmental and ecological causes that drive viral spillover? Third, how does virus spillover occur from bats and rodents to humans?

Mixed Effects of Habitat Degradation and Resources on Hantaviruses in Sympatric Wild Rodent Reservoirs within a Neotropical Forest.

Understanding the ecology of rodent-borne hantaviruses is critical to assessing the risk of spillover to humans. Longitudinal surveys have suggested that hantaviral prevalence in a given host population is tightly linked to rodent ecology and correlates with changes in the species composition of a rodent community over time and/or habitat composition. We tested two hypotheses to identify whether resource addition and/or habitat composition may affect hantavirus prevalence among two sympatric reservoir hosts in a neotropical forest: (i) increased food resources will alter the rodent community and thus hantaviral prevalence; and (ii) host abundance and viral seroprevalence will be associated with habitat composition. We established a baseline of rodent-virus prevalence in three grid pairs of distinct habitat compositions and subjected one grid of each pair to resource augmentation. Increased rodent species diversity was observed on grids where food was added versus untreated control grids during the first post-treatment sampling session. Resource augmentation changed species community composition, yet it did not affect the prevalence of hantavirus in the host population over time, nor was there evidence of a dilution effect. Secondly, we show that the prevalence of the virus in the respective reservoir hosts was associated with habitat composition at two spatial levels, independent of resource addition, supporting previous findings that habitat composition is a primary driver of the prevalence of hantaviruses in the neotropics.

Sympatry and habitat associations of sigmodontine rodents in a neotropical forest-savanna interface.

Small mammal communities in the Neotropics are composed largely of sigmodontine rodents. However, many questions regarding these communities remain unanswered, especially those pertaining to fine-scale sympatry and habitat selection. To address this, we examined sigmodontine community structure and vegetation in the western margin of the Upper Paraná Atlantic Forest and the southwestern-most extent of the Cerrado (CE) (an extensive South American savanna ecoregion) of Paraguay. Vegetation classifications were derived from satellite imagery combined with maps based on extensive ground-based surveys. The three most abundant species (Akodon montensis, Hylaeamys megacephalus, and Oligoryzomys nigripes) were found most often in microsympatry with conspecifics, and were negatively associated with other species. Akodon montensis was associated with high forest (HF), and H. megacephalus with bamboo understory (BU), whereas O. nigripes did not exhibit a habitat preference. The first two species' distributions within the landscape were found to be driven primarily by habitat selection, and O. nigripes by a behavioral response (avoidance) to the presence of the other two species. Moreover, habitat influences whether or not a particular species associates with, or avoids, conspecifics or other species.

Sigmodontine community and species responses to El Niño and precipitation in different levels of forest degradation.

Few studies have focused on rodent communities at the margins of an ecoregion or the limits of species' distributions, where the community may be more sensitive to extrinsic variables, both biotic and abiotic. This study evaluates sigmodontine rodent species diversity and overall abundance, and variation associated with climatic variables, in three locations with differing levels of habitat degradation. The study was conducted in northeastern Paraguay, near the western limit of the Upper Paraná Atlantic Forest and near the distributional limits of the three most abundant species in the study sites. Three mark-recapture grids were established and classified as least, moderately and most-degraded based on an analysis of several vegetation parameters. The grids were sampled for five consecutive nights, six times during two years. Shannon diversity and overall abundance were calculated for each sample. Monthly Multivariate ENSO Index and rainfall values were obtained from publicly available resources. Product-moment correlations were calculated between community and climatic parameters, including cumulative values for the climatic variables. The same correlations were calculated for the three common sigmodontine species. 1,632 captures were recorded, representing 13 sigmodontine species. Species richness in the samples (one session on one grid) varied from four to seven. Akodon montensis, Hylaeamys megacephalus and Oligoryzomys nigripes were the three most abundant species. In general, species diversity was negatively correlated with ENSO index, precipitation and precipitation anomaly, including cumulative one- to six-month cumulative values of each. Total sigmodontine abundance was positively correlated with the climatic variables. However, these correlations were not uniform among the three levels of habitat degradation, nor did the three abundant species show similar correlation patterns. The three most abundant species are each near their distributional limits, whereas several less abundant species have distributions that extend well beyond the study area. This somewhat counterintuitive result bears further investigation in other sites at ecoregional margins, to determine whether it is a commonly observed pattern, or an exception. Overall sigmodontine abundances were generally reflective of Akodon montensis abundance, which generally correlated with precipitation (including cumulative amounts). Our analyses of these longitudinal data showed two major effects on sigmodontine species diversity and population. First, they are impacted by habitat and secondly, they are affected by climate (ENSO, precipitation). However, individual species are not impacted similarly. Akodon montensis abundances primarily were correlated with abiotic (climatic) variables, and the correlations were consistent across habitats (biotic factors). In contrast, Hylaeamys megacephalus abundance was correlated with climatic variables in two habitats, but not the moderately-degraded habitat, and Oligoryzomys nigripes abundance was not correlated with climate in the most-degraded habitat. Pocos estudios se han centrado en las comunidades de roedores en los márgenes de una ecorregión o en los límites de las distribuciones de las especies, donde la comunidad puede ser más sensible a las variables extrínsecas, tanto bióticas como abióticas. Este estudio evalúa la diversidad de especies y la abundancia general de roedores sigmodontinos, y la variación asociada con las variables climáticas, en tres lugares con diferentes niveles de degradación del hábitat. Se establecieron tres parcelas de captura-marca-recaptura y se clasificaron como la menos, moderada y más degradadas basadas en un análisis de vegetación de varios parámetros. Las parcelas fueron muestreadas durante cinco noches consecutivas, seis veces durante dos años. La diversidad de Shannon y la abundancia general se calcularon para cada muestra. El índice mensual de ENOS multivariable y los valores de precipitación se obtuvieron de sitios accesibles en Internet. Las correlaciones producto-momento se calcularon entre los parámetros climáticos y de la comunidad, incluidos los valores acumulados para las variables climáticas. Se calcularon las mismas correlaciones para las tres especies comunes de sigmodontinos. Se registraron 1,632 capturas, representando 13 especies sigmodontinos. La riqueza de especies en las muestras (una sesión en una parcela) varió de cuatro a siete. Akodon montensis, Hylaeamys megacephalus y Oligoryzomys nigripes fueron las tres especies más abundantes. En general, la diversidad de especies se correlacionó negativamente con el índice ENOS, la precipitación y la anomalía de la precipitación, incluidos los valores acumulativos de uno a seis meses de cada uno. La abundancia total de sigmodontinos se correlacionó positivamente con las variables climáticas. Sin embargo, estas correlaciones no fueron uniformes entre los tres niveles de degradación del hábitat, ni tampoco entre las tres especies abundantes. Las tres especies más abundantes están cada una cerca de sus límites de distribución, mientras que varias especies menos abundantes tienen distribuciones que se extienden mucho más allá de este sitio. Este resultado algo contraintuitivo conlleva una mayor investigación en otros sitios en los márgenes ecorregionales, para determinar si es un patrón observado comúnmente, o una excepción. Las abundancias de sigmodontinos generalmente reflejaron la abundancia de Akodon montensis, que generalmente se correlacionó con la precipitación (incluidas las cantidades acumuladas). Las conclusiones destacadas de este estudio fueron: (1) diferentes niveles de degradación del hábitat se correlacionan con la variación en la diversidad de especies y la abundancia general de los sigmodontinos, y las especies individuales no muestran los mismos niveles de correlación entre los diferentes hábitats; y (2) la variabilidad climática (ENOS y precipitación) también afecta la diversidad de especies sigmodontinos y la abundancia de la población, y las especies comunes no muestran correlaciones similares entre sí. Las abundancias de Akodon montensis se correlacionaron principalmente con variables abióticas (climáticas), y las correlaciones fueron consistentes en todos los hábitats (factores bióticos). En contraste, la abundancia de Hylaeamys megacephalus se correlacionó con las variables climáticas en dos hábitats, pero no en el hábitat moderadamente degradado, y Oligoryzomys nigripes no se correlacionó con el clima en el hábitat más degradado.

Habitat, species richness and hantaviruses of sigmodontine rodents within the Interior Atlantic Forest, Paraguay.

Four of the nine sigmodontine tribes have species that serve as reservoirs of rodent-borne hantaviruses (RBO-HV), few have been studied in any depth. Several viruses have been associated with human cases of hantavirus pulmonary syndrome often through peridomestic exposure. Jabora (JABV) and Juquitiba (JUQV), harbored by Akodon montensis and Oligoryzomys nigripes, respectively, are endemic and sympatric in the Reserva Natural de Bosque Mbaracayú (RNBM), Paraguay, a protected area of the Interior Atlantic Forest. Rodent communities were surveyed along a 30 km stretch of the RNBM in eight vegetation classifications (Low, High, Bamboo, Riparian and Liana Forests, Bamboo Understory, Cerrado, and Meadow/Grasslands). We collected 417 rodents from which 11 species were identified; Akodon montensis was the predominant species (72%; 95%CI: 64.7%-76.3%), followed by Hylaeamys megacephalus (15% (11.2%-18.2%)) and Oligoryzomys nigripes (9% (6.6%-12.4%)). We examined the statistical associations among habitat (vegetation class) type, rodent species diversity, population structure (age, sex, and weight), and prevalence of RBO-HV antibody and/or viral RNA (Ab/RNA) or characteristic Leishmania tail lesions. Ab/RNA positive rodents were not observed in Cerrado and Low Forest. A. montensis had an overall Ab/RNA prevalence of 7.7% (4.9%-11.3%) and O. nigripes had an overall prevalence of 8.6% (1.8%-23.1%). For A. montensis, the odds of being Ab/RNA positive in High Forest was 3.73 times of the other habitats combined. There was no significant difference among age classes in the proportion of Ab/RNA positive rodents overall (p = 0.66), however, all 11 RNA-positive individuals were adult. Sex and habitat had independent prognostic value for hantaviral Ab/RNA in the study population; age, presence of tail scar/lesion (19% of the rodents) and weight did not. Adjusting for habitat, female rodents had less risk of becoming infected. Importantly, these data suggest habitat preferences of two sympatric rodent reservoirs for two endemic hantaviruses and the importance of including habitat in models of species diversity and habitat fragmentation.

COMPOSITION AND CHARACTERISTICS OF A DIVERSE DIDELPHID COMMUNITY (MAMMALIA: DIDELPHIMORPHIA) IN SUB-TROPICAL SOUTH AMERICA.

Few studies have reported didelphid communities of ≥10 species, and all of these have been from within the tropics sensu stricto of South America. Herein a community of 12 species of didelphids is described from a sub-tropical site in south-central South America. The Reserva Natural del Bosque Mbaracayú, in northeastern Paraguay, lies at the western margin of the Interior Atlantic Forest and the southwestern limit of the Cerrado, two important South American ecoregions. The rich didelphid community in this area likely results from the mosaic of habitats encountered at the distributional limits of these two ecoregions. Within the context of this mosaic, the species' habitat associations and vertical occupancy are discussed, as well as the reproductive patterns and population abundance variation of the more commonly encountered species. Three Monodelphis species were found in sympatry, all strictly terrestrial, along with Cryptonanus chacoensis. Marmosa paraguayana shared all habitats with Gracilinanus agilis, and both of these species primarily were arboreal. Although this natural reserve has been more extensively sampled than any other area in Paraguay, numerous questions remain unanswered regarding this rich didelphid community.

Pocos estudios han registrado comunidades de didélfidos de ≥10 especies, y todas ellas han sido de los trópicos sensu stricto de América del Sur. Aquí reportamos una comunidad de 12 especies de didélfidos de un sitio subtropical en el centro sur de América del Sur. La Reserva Natural del Bosque Mbaracayú, en el noreste de Paraguay, se encuentra en el margen occidental del Bosque Atlántico Interior, y el límite suroeste del Cerrado, dos ecorregiones sudamericanas importantes. La rica comunidad de didélfidos en esta área es probablemente el resultado del mosaico de hábitats encontrados en los límites de distribución de estas dos ecorregiones. Dentro del contexto de este mosaico, discutimos las asociaciones de hábitats y la ocupación vertical de las especies, los patrones reproductivos y la variación de la abundancia de las poblaciones de las especies más comúnmente encontradas. Encontramos tres especies de Monodelphis en simpatría, y todas estrictamente terrestres, junto con Cryptonanus chacoensis. Marmosa paraguayana compartió todos sus hábitats con Gracilinanus agilis, y ambas especies eran principalmente arbóreas. Enfatizamos que a pesar de que esta reserva natural ha sido muestreada más extensamente que cualquier otra área en Paraguay, muchas preguntas siguen sin respuesta con respecto a esta rica comunidad de didélfidos.

Morphometric variation in Periglischrus torrealbai (Acari: Spinturnicidae) on three species of host bats (Chiroptera: Phyllostomidae) with a new record of host species.

We evaluated morphometric variation of the mite Periglischrus torrealbai (Spinturnicidae) on three species of host bats: Phyllostomus discolor, P. hastatus, and Tonatia bidens (Phyllostomidae). A total of 67 females and 74 males of P. torrealbai were collected from 41 host individuals of these three bat species that were sampled in Brazil, Paraguay, and Peru. Twenty-one measurements from the dorsal side and 28 from the ventral side were recorded from female mites and 21 dorsal and 34 ventral measurements were taken from males. To evaluate morphological variation of P. torrealbai on different species of host bats, principal component analysis and unweighted pair-group method using arithmetic averages cluster analysis with Euclidean distances were used. Both analyses showed three groups of mites clearly separated: group 1 comprised all ectoparasites collected from T. bidens, group 2 included all mites from P. hastatus, and group 3 had all those from P. discolor. This result indicates that P. torrealbai varies morphologically by host bat species and suggests that this nominal species comprises three morphologically distinct species. In the present study, we record for the first time, the association between P. torrealbai and T. bidens. Our data reinforce the high relationship of specificity between Periglischrus mites and phyllostomid bat species.

Ecology of rodent-associated hantaviruses in the Southern Cone of South America: Argentina, Chile, Paraguay, and Uruguay.

Thirteen hantavirus genotypes, associated with at least 12 sigmodontine reservoir rodents, have been recognized in the four countries that represent the Southern Cone of South America. Host-virus relationships are not as well defined as in North America; several Southern Cone hantaviruses appear to share a common host and some viruses do not occur throughout the range of their host. Although hantavirus-host relationships in the Southern Cone are less strictly concordant with the single-host-single-virus pattern reported elsewhere, recent studies suggest that much of the ambiguity may result from an incomplete understanding of host and hantavirus systematics. Although some Southern Cone host species are habitat generalists, some sympatric species are habitat specialists, helping to explain how some strict host-virus pairings may be maintained. In some cases, host population densities were higher in peridomestic habitats and prevalence of hantavirus infection was higher in host populations in peridomestic habitats. Seasonal and multiyear patterns in climate and human disturbance affect host population densities, prevalence of infection, and disease risk to humans. Unusually high hantavirus antibody prevalence in indigenous human populations may be associated with frequent and close contact with host rodents. Ongoing studies are improving our understanding of hantavirus-host ecology and providing tools that may predict human risk.

Phylogenetic exploration of hantaviruses in Paraguay reveals reassortment and host switching in South America.

BACKGROUND: Longitudinal mark-recapture studies of rodents in two sites in the Mbaracayú Biosphere Reserve in the Interior Atlantic Forest of eastern Paraguay have revealed a complex and intriguing pattern of hantaviruses harbored by rodents in this area. Full-length sequencing and phylogenetic analyses were conducted for several rodents from Akodon montensis and Oligoryzomys fornesi. The phylogenetic relationships of these viruses were analyzed in the context of hantaviruses in South America with published S- and M-segment sequences. FINDINGS: Phylogenetic analyses of hantaviruses identified in the Mbaracayú Biosphere Reserve in Paraguay revealed Jabora and Juquitiba viruses are harbored by Akodon montensis and Oligoryzomys fornesi, respectively. These analyses revealed that in general the constituents of the major subclade for the S- and M-segments differ for the South American hantaviruses. Further, the two major groups within subclade C for the M-segment reflect in general the lethality associated with the viruses within each group. CONCLUSIONS: Phylogenetic studies of Jabora and Juquitiba viruses and other Paraguayan viruses in the context of American hantaviruses revealed reassortment and host-switching in the evolution of South American hantaviruses.

Novel use of intraoperative dexmedetomidine infusion for sedation during spinal cord stimulator lead placement via surgical laminectomy.

BACKGROUND: Spinal cord stimulators are most often placed through a percutaneous approach using minimal sedation and local anesthesia to facilitate intraoperative testing. However, when leads need to be placed using a laminectomy incision additional anesthesia is required which can complicate intraoperative testing. There is no consensus as to the best anesthetic choice when laminectomy-placed leads are required. OBJECTIVE: We present 2 cases where spinal cord stimulator leads were implanted through a surgical laminectomy under sedation using dexmedetomidine infusion and local anesthesia to provide a cooperative patient for intraoperative testing. CASE REPORT: Patient #1: A 40-year-old female with Complex Regional Pain Syndrome secondary to an automobile accident who had good pain control with a spinal cord stimulator until a lead fracture resulted in loss of stimulation. She required a laminectomy-placed lead which was implanted under dexmedetomidine infusion and local anesthesia. Patient #2: A 54-year-old female with Failed Back Syndrome who had good pain control until a lead fracture resulted in loss of stimulation. She underwent a laminectomy-placed lead, new battery pocket, and removal of the old system under a dexmedetomidine infusion and local anesthesia. LIMITATIONS: Report of only 2 cases. CONCLUSIONS: The anesthetic management from a laminectomy-placed spinal cord stimulator can present a difficult choice. A general anesthetic or even deep sedation can provide good operative conditions but limits intraoperative testing or in the case of deep sedation risks losing the airway in the prone position. On the other hand, minimal sedation, which facilitates intraoperative testing, can make the surgical procedure extremely uncomfortable or even unbearable. Dexmedetomidine infusion and local anesthesia provide sedation for the operative portions while rendering the patient alert and cooperative during intraoperative testing.

Sympatry of 2 hantavirus strains, paraguay, 2003-2007.

To explore geographic and host-taxonomic patterns of hantaviruses in Paraguay, we established sampling sites in the Mbaracayu Biosphere Reserve. We detected Jabora virus and Itapua37/Juquitiba-related virus in locations approximately 20 m apart in different years, which suggested sympatry of 2 distinct hantaviruses.

A habitat-based model for the spread of hantavirus between reservoir and spillover species.

New habitat-based models for spread of hantavirus are developed which account for interspecies interaction. Existing habitat-based models do not consider interspecies pathogen transmission, a primary route for emergence of new infectious diseases and reservoirs in wildlife and man. The modeling of interspecies transmission has the potential to provide more accurate predictions of disease persistence and emergence dynamics. The new models are motivated by our recent work on hantavirus in rodent communities in Paraguay. Our Paraguayan data illustrate the spatial and temporal overlaps among rodent species, one of which is the reservoir species for Jabora virus and others which are spillover species. Disease transmission occurs when their habitats overlap. Two mathematical models, a system of ordinary differential equations (ODE) and a continuous-time Markov chain (CTMC) model, are developed for spread of hantavirus between a reservoir and a spillover species. Analysis of a special case of the ODE model provides an explicit expression for the basic reproduction number, R(0), such that if R(0)<1, then the pathogen does not persist in either population but if R(0)>1, pathogen outbreaks or persistence may occur. Numerical simulations of the CTMC model display sporadic disease incidence, a new behavior of our habitat-based model, not present in other models, but which is a prominent feature of the seroprevalence data from Paraguay. Environmental changes that result in greater habitat overlap result in more encounters among various species that may lead to pathogen outbreaks and pathogen establishment in a new host.

Intracerebroventricular administration of drugs.

Intracerebroventricular drug administration is a method that bypasses the blood-brain barrier and other mechanisms that limit drug distribution into the brain, allowing high drug concentrations to enter the central compartment. Instillation of drugs directly into the ventricles of the brain must be done carefully and with full consideration of factors affecting the efficacy and safety of this route of administration. These factors include the osmolarity, pH, volume, and presence of preservatives and diluents of the drug solution being administered. Very few studies have formally investigated intraventricular therapies, and dosing recommendations may vary widely depending on the agent and the patient. Many antimicrobials have been given intraventricularly, although very few prospective studies have evaluated this strategy. There are wide variations among the reports regarding dosage regimens and the pharmacokinetics of the antimicrobials used. Guidance on appropriate formulations and their use is lacking. Clinicians should be aware of their patients' ongoing disease processes and neurologic status, as well as pertinent physiochemical properties of drugs when formulating them for intracerebroventricular administration; a high index of suspicion should be maintained when monitoring patients for adverse drug events after instillation.

Microhabitat characteristics of Akodon montensis, a reservoir for hantavirus, and hantaviral seroprevalence in an Atlantic forest site in eastern Paraguay.

Hantaviruses may cause serious disease when transmitted to humans by their rodent hosts. Since their emergence in the Americas in 1993, there have been extensive efforts to understand the role of environmental factors on the presence of these viruses in their host rodent populations. HPS outbreaks have been linked to precipitation, but climatic factors alone have not been sufficient to predict the spatial-temporal dynamics of the environment-reservoir-virus system. Using a series of mark-recapture sampling sites located at the Mbaracayú Biosphere Reserve, an Atlantic Forest site in eastern Paraguay, we investigated the hypothesis that microhabitat might also influence the prevalence of Jaborá hantavirus within populations of its reservoir species, Akodon montensis. Seven trapping sessions were conducted during 2005-2006 at four sites chosen to capture variable microhabitat conditions within the study site. Analysis of microhabitat preferences showed that A. montensis preferred areas with little forest overstory and denser vegetation cover on and near the ground. Moreover, there was a significant difference in the microhabitat occupied by antibody-positive vs antibody-negative rodents, indicating that microhabitats with greater overstory cover may promote transmission and maintenance of hantavirus in A. montensis.

Intrathecal granuloma in a patient receiving high dose hydromorphone.

Intrathecal granuloma formation has commonly been described with morphine therapy. It has been suggested that a high concentration of intrathecal morphine may be responsible for this complication. Much less commonly, intrathecal hydromorphone has been associated with intrathecal granuloma formation. In the current case we report the evaluation and management of an intrathecal granuloma in a patient receiving a relatively high concentration of intrathecal hydromorphone. A nonsurgical, conservative approach to management involves stopping the infusion and observing the patient for improvement as the granuloma mass often slowly resolves once the infusion is stopped. Cessation of the infusion or addition of clonidine to the IDDS admixture in conjunction with close clinical monitoring may be reasonable treatment options in patients with an asymptomatic or mildly symptomatic inflammatory mass. In the current study, rapidly declining neurologic function with a confirmed inflammatory mass adherent to the spinal canal necessitated urgent surgical intervention. Though use of intrathecal hydromorphone still represents an off label application, this opiate is commonly employed as an alternative first line analgesic agent. This case report highlights the potential of high-dose and high infusate concentration intrathecal hydromorphone to form an inflammatory granuloma.

Genetic characterization and phylogeny of a hantavirus from Western Mexico.

Hantaviruses can cause two serious illnesses when transmitted from their rodent reservoirs to humans; hantavirus pulmonary syndrome (HPS) in the New World and hemorrhagic fever with renal syndrome in the Old World. Cases of HPS were first recognized in the Americas in small, focal outbreaks in rural populations in the Southwestern USA in 1993. Since that time, outbreaks as well as sporadic cases of HPS have been recognized throughout the Americas. Remarkably, HPS cases have not been reported in Mexico. Mexico is one of the most biodiverse regions in the world and this is reflected in the species diversity of the peromyscine, sigmodontine and oryzomyine rodents; all potential hosts of hantaviruses. Hence, we collected and surveyed several rodent species in Western Mexico and identified three previously unrecognized rodents with antibodies to hantaviral antigens: Oryzomys couesi, Sigmodon mascotensis and Baiomys musculus. The S and M segments cloned from O. couesi and S. mascotensis, referred to herein as Playa de Oro (ORO) virus, showed strongest similarity to Bayou and Catacamas viruses with 92/93% and 92/92% similarity based on S/M amino acid sequences, respectively. This and phylogenetic analysis of the M and S segments suggests that ORO virus is a unique genotype within Hantavirus.

Phylogenetic and geographical relationships of hantavirus strains in eastern and western Paraguay.

Recently, we reported the discovery of several potential rodent reservoirs of hantaviruses in western (Holochilus chacarius) and eastern Paraguay (Akodon montensis, Oligoryzomys chacoensis, and O. nigripes). Comparisons of the hantavirus S- and M-segments amplified from these four rodents revealed significant differences from each another and from other South American hantaviruses. The ALP strain from the semiarid Chaco ecoregion clustered with Leguna Negra and Rio Mamore (LN/RM), whereas the BMJ-NEB strain from the more humid lower Chaco ecoregion formed a clade with Oran and Bermejo. The other two strains, AAI and IP37/38, were distinct from known hantaviruses. With respect to the S-segment sequence, AAI from eastern Paraguay formed a clade with ALP/LN/RM, but its M-segment clustered with Pergamino and Maciel, suggesting a possible reassortment. AAI was found in areas experiencing rapid land cover fragmentation and change within the Interior Atlantic Forest. IP37/38 did not show any strong association with any of the known hantavirus strains.

A longitudinal study of Bayou virus, hosts, and habitat.

What is currently known about the ecology of North American hantaviruses has come largely from studies on Sin Nombre virus (SNV). We conducted a longitudinal study of Bayou virus (BAYV), the second-leading agent of hantavirus pulmonary syndrome in the United States. Antibodies to hantavirus were detected from Oryzomys palustris (most commonly infected species), Sigmodon hispidus, Peromyscus leucopus, Reithrodontomys fulvescens, and Baiomys taylori. However, only O. palustris had viral RNA in tissues and excreta, suggesting that antibodies detected in other species may have resulted from spill-over infection. Seroprevalence rates averaged around 16% for O. palustris and varied seasonally. The heaviest males exhibited the highest levels of seroprevalence. Seroprevalence was higher in coastal prairie (20.0%) than old-fields (10.5%) and was associated with host abundance. These patterns are similar to those of SNV and can be used in identification of potentially at-risk areas.