Antibodies to Influenza and West Nile Viruses in Horses in Mexico

INFLUENZA A virus (IAV) (family Orthomyxoviridae) is a highly infectious respiratory pathogen of birds and mammals, including human beings and horses (Palese and Shaw 2007). The virus is classified into different subtypes based on the antigenic properties of the haemagglutinin (HA) and neuraminidase (NA) proteins. Sixteen HA subtypes (H1 to H16) and nine NA subtypes (N1 to N9) have been identified (Fouchier and others 2005). Two subtypes, H3N8 and H7N7, have been isolated from horses. The H7N7 subtype was first isolated from a horse in Czechoslovakia in 1956 (Prague/56) (Sovinova and others 1958), and the H3N8 subtype was first isolated from a horse in Miami, USA, in 1963 (Waddell and others 1963). The H7N7 subtype has not been isolated from horses for three decades and is presumed to be extinct (Webster 1993). The H3N8 subtype is currently a common cause of disease in horses worldwide. In horses, influenza is characterized by an abrupt onset of pyrexia, depression, coughing and nasal discharge, and is often complicated by secondary bacteria infections that can lead to pneumonia and death (Hannant and Mumford 1996). Although H3N8 is a major cause of morbidity in horses throughout the world, information on the seroprevalence of IAV in horses and other domestic animals in Mexico is limited.

Acetaminophen as an Oral Toxicant for Nile Monitor Lizards (Varanus niloticus) and Burmese Pythons (Python molurus bivittatus)

Context. Invasive species are a growing global problem. Biological invasions can result in numerous harmful impacts on local ecologies, and non-native herpetofauna are frequently ignored. Nile monitor lizards (Varanus niloticus) and Burmese pythons (Python molurus bivittatus, recently reassessed as Python bivittatus bivittatus), have become established in southern Florida. Both are large, semi-aquatic predators that pose serious threats to a variety of threatened and endangered species, as well as to the unique ecology of the area. Aims. Acetaminophen (CAS#103-90-2), a lethal oral toxicant for the invasive brown treesnake (Boiga irregularis) on Guam, was investigated as a possible toxicant in juvenile Burmese pythons and Nile monitors. Methods. Dead neonatal mouse (DNM) baits containing 0, 10, 20, or 40 mg acetaminophen were force-fed to Nile monitors, whereas DNM containing doses of 0, 20, 40, or 80 mg were freely consumed by Burmese pythons. Subjects were frequently observed post-treatment for general condition and position, with special attention paid to activity (if any), behaviour, respiration, bleeding, emesis, ataxia, and mortality. Key results. In Nile monitors, acetaminophen doses of 10, 20, or 40 mg resulted in 0, 50 and 100% mortality, respectively. In Burmese pythons, doses of 20, 40, or 80 mg resulted in 14.3, 85.7 and 100% mortality, respectively. No mortality was observed in control individuals of either species. A negative correlation between dosage (mg kg–1) and time-to-death was observed in both species. Dosages ranging from 522 to 2438 mg kg–1 and 263 to 703 mg kg–1 were uniformly lethal to monitors and pythons, respectively. Neither species exhibited signs of pain or discomfort following acetaminophen treatment. Conclusions. Acetaminophen is an effective toxicant in juvenile Nile monitors and Burmese pythons. Further investigation into acetaminophen toxicity in adults of these species is merited. Implications. Although further investigation into adult lethal dosages and strategies to optimize bait deployment while minimizing secondary hazards is required, acetaminophen may have a role to play in the control of these invasive species in Florida.

Seeking a Second Opinion: Uncertainty in Disease Ecology

Analytical methods accounting for imperfect detection are often used to facilitate reliable inference in population and community ecology. We contend that similar approaches are needed in disease ecology because these complicated systems are inherently difficult to observe without error. For example, wildlife disease studies often designate individuals, populations, or spatial units to states (e.g., susceptible, infected, post-infected), but the uncertainty associated with these state assignments remains largely ignored or unaccounted for. We demonstrate how recent developments incorporating observation error through repeated sampling extend quite naturally to hierarchical spatial models of disease effects, prevalence, and dynamics in natural systems. A highly pathogenic strain of avian influenza virus in migratory waterfowl and a pathogenic fungus recently implicated in the global loss of amphibian biodiversity are used as motivating examples. Both show that relatively simple modifications to study designs can greatly improve our understanding of complex spatio-temporal disease dynamics by rigorously accounting for uncertainty at each level of the hierarchy.

Landscape Genetics of Raccoons (Procyon lotor) Associated with Ridges and Valleys of Pennsylvania: Implications for Oral Rabies Vaccination Programs

Raccoons are the reservoir for the raccoon rabies virus variant in the United States. To combat this threat, oral rabies vaccination (ORV) programs are conducted in many eastern states. To aid in these efforts, the genetic structure of raccoons (Procyon lotor) was assessed in southwestern Pennsylvania to determine if select geographic features (i.e., ridges and valleys) serve as corridors or hindrances to raccoon gene flow (e.g., movement) and, therefore, rabies virus trafficking in this physiographic region. Raccoon DNA samples (n = 185) were collected from one ridge site and two adjacent valleys in southwestern Pennsylvania (Westmoreland, Cambria, Fayette, and Somerset counties). Raccoon genetic structure within and among these study sites was characterized at nine microsatellite loci. Results indicated that there was little population subdivision among any sites sampled. Furthermore, analyses using a model-based clustering approach indicated one essentially panmictic population was present among all the raccoons sampled over a reasonably broad geographic area (e.g., sites up to 36 km apart). However, a signature of isolation by distance was detected, suggesting that widths of ORV zones are critical for success. Combined, these data indicate that geographic features within this landscape influence raccoon gene flow only to a limited extent, suggesting that ridges of this physiographic system will not provide substantial long-term natural barriers to rabies virus trafficking. These results may be of value for future ORV efforts in Pennsylvania and other eastern states with similar landscapes.

A Silent Enzootic of an Orthopoxvirus in Ghana, West Africa: Evidence for Multi-Species Involvement in the Absence of Widespread Human Disease

Human monkeypox has never been reported in Ghana, but rodents captured in forested areas of southern Ghana were the source of the monkeypox virus introduced into the United States in 2003. Subsequent to the outbreak in the United States, 204 animals were collected from two commercial trapping sites in Ghana. Animal tissues were examined for the presence of orthopoxvirus (OPXV) DNA using a real-time polymerase chain reaction, and sera were assayed for antibodies against OPXV. Animals from five genera (Cricetomys , Graphiurus , Funiscirus, and Heliosciurus ) had antibodies against OPXV, and three genera (Cricetomys , Graphiurus , and Xerus) had evidence of OPXV DNA in tissues. Additionally, 172 persons living near the trapping sites were interviewed regarding risk factors for OPXV exposure, and their sera were analyzed. Fifty-three percent had IgG against OPXV; none had IgM. Our findings suggest that several species of forest-dwelling rodents from Ghana are susceptible to naturally occurring OPXV infection, and that persons living near forests may have low-level or indirect exposure to OPXV-infected animals, possibly resulting in sub-clinical infections.

Invasive Litter, Not an Invasive Insectivore, Determines Invertebrate Communities in Hawaiian Forests

In Hawaii, invasive plants have the ability to alter litter-based food chains because they often have litter traits that differ from native species. Additionally, abundant invasive predators, especially those representing new trophic levels, can reduce prey. The relative importance of these two processes on the litter invertebrate community in Hawaii is important, because they could affect the large number of endemic and endangered invertebrates. We determined the relative importance of litter resources, represented by leaf litter of two trees, an invasive nitrogen-fixer, Falcataria moluccana, and a native tree, Metrosideros polymorpha, and predation of an invasive terrestrial frog, Eleutherodactylus coqui, on leaf litter invertebrate abundance and composition. Principle component analysis revealed that F. moluccana litter creates an invertebrate community that greatly differs from that found in M. polymorpha litter. We found that F. moluccana increased the abundance of non-native fragmenters (Amphipoda and Isopoda) by 400% and non-native predaceous ants (Hymenoptera: Formicidae) by 200%. E. coqui had less effect on the litter invertebrate community; it reduced microbivores by 40% in F. moluccana and non-native ants by 30% across litter types. E. coqui stomach contents were similar in abundance and composition in both litter treatments, despite dramatic differences in the invertebrate community. Additionally, our results suggest that invertebrate community differences between litter types did not cascade to influence E. coqui growth or survivorship. In conclusion, it appears that an invasive nitrogen-fixing tree species has a greater influence on litter invertebrate community abundance and composition than the invasive predator, E. coqui.

Eight Polymorphic Microsatellite Loci Developed and Characterized From Townsend’s Big-Eared Bat, Corynorhinus Townsendii

Two of the five subspecies of the western big-eared bat, Corynorhinus townsendii, are listed as federally endangered with the remaining three being of conservation concern. Knowing the degree of connectivity among populations would aid in the establishment of sound conservation and management plans for this taxon. For this purpose, we have developed and characterized eight polymorphic microsatellite markers.

Development of 10 Polymorphic Microsatellite Loci Isolated From The Mountain Beaver, Aplodontia rufa rufa (Rafinesque)

We developed 10 microsatellite markers for the mountain beaver, Aplodontia rufa rufa. In three populations of A. r. rufa, the number of alleles for these loci ranged from monomorphic to nine. Average observed heterozygosities in these populations ranged from 0.29 to 0.60. We also tested previously published markers from the endangered subspecies A. r. nigra in A. r. rufa populations.

Development and Characterization of 15 Polymorphic Microsatellite Loci Isolated From Rafinesque’s Big-Eared Bat, Corynorhinus rafinesquii

We developed and characterized 15 microsatellite markers for Rafinesque’s big-eared bat, Corynorhinus rafinesquii. In a population from Tennessee, the number of alleles per locus ranged from three to 13 and observed heterozygosities were 0.35 to 0.97 per locus. These loci will provide appropriate variability for estimation of population connectivity, demographic parameters, and genetic diversity for this species of concern.

Intraspecific Comparison of Population Structure, Genetic Diversity, and Dispersal Among Three Subspecies of Townsend’s Big-Eared Bats, Corynorhinus townsendii townsendii, C. t. pallescens, and the Endangered C. t. birginianus

Townsend’s big-eared bat, Corynorhinus townsendii, is distributed broadly across western North America and in two isolated, endangered populations in central and eastern United States. There are five subspecies of C. townsendii; C. t. pallescens, C. t. australis, C. t. townsendii, C. t. ingens, and C. t. virginianus with varying degrees of concern over the conservation status of each. The aim of this study was to use mitochondrial and microsatellite DNA data to examine genetic diversity, population differentiation, and dispersal of three C. townsendii subspecies. C. t. virginianus is found in isolated populations in the eastern United States and was listed as endangered under the Endangered Species Act in 1979. Concern also exists about declining populations of two western subspecies, C. t. pallescens and C. t. townsendii. Using a comparative approach, estimates of the genetic diversity within populations of the endangered subspecies, C. t. virginianus, were found to be significantly lower than within populations of the two western subspecies. Further, both classes of molecular markers revealed significant differentiation among regional populations of C. t. virginianus with most genetic diversity distributed among populations. Genetic diversity was not significantly different between C. t. townsendii and C. t. pallescens. Some populations of C. t. townsendii are not genetically differentiated from populations of C. t. pallescens in areas of sympatry. For the western subspecies gene flow appears to occur primarily through male dispersal. Finally, geographic regions representing significantly differentiated and genetically unique populations of C. townsendii virginianus are recognized as distinct evolutionary significant units.

Experimental Infection of Richardson’s Ground Squirrels (Spermophilus Richardsonii ) With Attenuated and Virulent Strains pf Brucella abortus

A previous investigation of the safety of Brucella abortus strain RB51 (sRB51) in various nontarget species suggested that Richardson’s ground squirrels (Spermophilus richardsonii) may develop persistent infections when orally inoculated with the vaccine. In the present study, sRB51, B. abortus strain 19 (s19), and virulent B. abortus strain 9941 (s9941) were administered orally to Richardson’s ground squirrels to further characterize B. abortus infection in this species. Six groups of nongravid ground squirrels were orally inoculated with 6x108 colony forming units (cfu) sRB51 (n=10), 2.5x104 cfu s19 (n=10), 2.5x107 cfu s19 (n=6), 1.3x106 cfu s9941 (n=5), 2.1x108 cfu s9941 (n=5), or vaccine diluent (control; n=4). One of five animals in the lower-dose s19 group and two of three animals in the higher-dose s19 group showed persistence of bacteria in various tissues at 14 wk post-inoculation (PI). At 18 wk PI, one of five animals in the sRB51 group and one of five animals in the high-dose s9941 group were culture positive. Although we did detect some persistence of B. abortus strains at 18 wk, we found no evidence of pathology caused by B. abortus strains in nonpregnant Richardson’s ground squirrels based on clinical signs, gross lesions, and microscopic lesions.

Efficacy of Oral and Parenteral Routes of Mycobacterium bovis Bacille Calmette-Guerin Vaccination Against Experimental Bovine Tuberculosis in White-Tailed Deer (Odocoileus virginianus): A Feasibility Study

We investigated the efficacy of oral and parenteral Mycobacterium bovis bacille Calmette-Guerin Danish strain 1331 (BCG) in its ability to protect white-tailed deer (Odocoileus virginianus) against disease caused by M. bovis infection. Twenty-two white-tailed deer were divided into four groups. One group (n=5) received 109 colony-forming units (cfu) BCG via a lipid-formulated oral bait; one group (n=5) received 109 cfu BCG in culture directly to the oropharynx, one group (n=6) was vaccinated with 106 cfu BCG subcutaneously, and one group served as a control and received culture media directly to the oropharynx (n=6). All animals were challenged 3 mo after vaccination. Five months postchallenge the animals were examined for lesions. Results indicate that both oral forms of BCG and parenterally administerd BCG offered significant protection against M. bovis challenge as compared to controls. This study suggests that oral BCG vaccination may be a feasible means of controlling bovine tuberculosis in wild white-tailed deer populations.

Host-Specificity of Myxoma Virus: Pathogenesis of South American and North American Strains of Myxoma Virus in Two North American Lagomorph Species

The pathogenesis of South American and North American myxoma viruses was examined in two species of North American lagomorphs, Sylvilagus nuttallii (mountain cottontail) and Sylvilagus audubonii (desert cottontail) both of which have been shown to have the potential to transmit the South American type of myxoma virus. Following infection with the South American strain (Lausanne, Lu), S. nuttallii developed both a local lesion and secondary lesions on the skin. They did not develop the classical myxomatosis seen in European rabbits (Oryctolagus cuniculus). The infection at the inoculation site did not resolve during the 20-day time course of the trial and contained transmissible virus titres at all times. In contrast, S. audubonii infected with Lu had very few signs of disseminated infection and partially controlled virus replication at the inoculation site. The prototype Californian strain of myxoma virus (MSW) was able to replicate at the inoculation site of both species but did not induce clinical signs of a disseminated infection. In S. audubonii, there was a rapid response to MSW characterized by a massive T lymphocyte infiltration of the inoculation site by day 5. MSW did not reach transmissible titres at the inoculation site in either species. This might explain why the Californian myxoma virus has not expanded its host-range in North America.

Benefit Cost Scenarios of Potential Oral Rabies Vaccination for Skunks in California

Scenario-based analyses were computed for benefits and costs linked with hypothetical oral rabies vaccination (ORV) campaigns to contain or eliminate skunk-variant rabies in skunks (Mephitis mephitis) in California, USA. Scenario 1 assumed baiting eight zones (43,388 km2 total) that comprised 73% of known skunk rabies locations in the state. Scenario 2 also assumed baiting these eight zones, but further assumed that added benefits would result from preventing the spread of skunk-variant rabies into Los Angeles County, USA. Scenarios assumed a fixed bait cost ($1.24 each) but varied campaigns (one, two and three annual ORV applications), densities of baits (37.5/km2, 75/km2 and 150/km2), levels of prevention (50%, 75%, and 100%), and contingency expenditures if rabies recurred (20%, 40%, and 60% of campaign costs). Prorating potential annual benefits during a 12-yr time horizon yielded benefit-cost ratios (BCRs) between 0.16 and 2.91 and between 0.34 and 6.35 for Scenarios 1 and 2, respectively. Economic issues relevant to potentially managing skunk-variant rabies with ORV are discussed.

Deer Guards and Bump Gates for Excluding White-Tailed Deer From Fenced Resources

White-tailed deer (Odocoileus virginianus) causing damage is a reoccurring theme in the realm of wildlife damage management, especially regarding human safety, disease transmission, and agricultural losses. Fences often are the only reliable long-term nonlethal means of controlling deer damage. The efficacy of fences, however, relies on their weakest link: human-operated gates. Although not overly time-consuming, the act of closing a gate appears to be a burden to individuals, resulting in open-access to an otherwise protected resource. We examined the efficacy of 2 alternatives to traditional gates to evaluate their potential to be used for excluding or containing deer. We evaluated a commercially available kit for mechanically opening and closing gates and a modified deer guard that resembles a common cattle guard but incorporates bearing-mounted rollers as cross members. The gate kit proved effective in restricting deer access to bait throughout the study, but, in supplemental evaluations, we observed excessive rates of functional failure. Deer guards reduced deer entry into exclosures, but efficacy declined with time as deer walked and jumped across guards. With some refining, both guards and gates have potential to be useful components of an integrated biosecurity strategy.