TAXONOMIC REASSESSMENT AND MORPHOLOGICAL REDESCRIPTION OF NEMATOSTRIGEA SERPENS ANNULATA (DIGENEA: STRIGEIDAE) FROM OSPREY IN NORTH AMERICA.

Digenean trematodes in the strigeid genus Nematostrigea are parasites of various birds, most often ospreys (Pandion haliaetus). Nematostrigea currently contains 2 species, Nematostrigea serpens and Nematostigea hepatica. Nematostrigea serpens is divided into 2 subspecies: N. serpens serpens from the Palearctic and Indomalayan realms and N. serpens annulata from the Nearctic realm. In the present work, we studied the type material of N. serpens annulata and collected new specimens from an osprey in Wisconsin close to the type locality. The original description and illustration of N. serpens annulata were incomplete. Herein we provide a detailed redescription and new illustrations of N. serpens annulata on the basis of the type material as well as newly collected and sequenced specimens. Nematostrigea serpens annulata has a distinct tegumental thickening near the mid-length of the genital cone, which is absent in its congeners. Partial sequences of the large ribosomal subunit (28S) DNA and cytochrome c oxidase subunit 1 (COI) mitochondrial DNA gene sequences were generated from our specimens of N. serpens annulata. Comparisons with previously published sequences of N. serpens serpens from the Palearctic demonstrated 2.2% difference in 28S and 3.7% in COI. On the basis of morphological and molecular comparisons, we elevate N. serpens annulata to species status and recognize it as Nematostrigea annulata Dubois and Rausch, 1948.

Associations among Habitat Characteristics and Meningeal Worm Prevalence in Eastern South Dakota, USA.

Few studies have evaluated how wetland and forest characteristics influence the prevalence of meningeal worm ( Parelaphostrongylus tenuis ) infection of deer throughout the grassland biome of central North America. We used previously collected, county-level prevalence data to evaluate associations between habitat characteristics and probability of meningeal worm infection in white-tailed deer ( Odocoileus virginianus ) across eastern South Dakota, US. The highest-ranked binomial regression model for detecting probability of meningeal worm infection was spring temperature + summer precipitation + percent wetland; weight of evidence (wi=0.71) favored this model over alternative models, though predictive capability was low (Receiver operating characteristic=0.62). Probability of meningeal worm infection increased by 1.3- and 1.6-fold for each 1-cm and 1-C increase in summer precipitation and spring temperature, respectively. Similarly, probability of infection increased 1.2-fold for each 1% increase in wetland habitat. Our findings highlight the importance of wetland habitat in predicting meningeal worm infection across eastern South Dakota. Future research is warranted to evaluate the relationships between climatic conditions (e.g., drought, wet cycles) and deer habitat selection in maintaining P. tenuis along the western boundary of the parasite.

Environmental Factors Influencing White-Tailed Deer (Odocoileus virginianus) Exposure to Livestock Pathogens in Wisconsin.

White-tailed deer (Odocoileus virginianus) are commonly exposed to disease agents that affect livestock but environmental factors that predispose deer to exposure are unknown for many pathogens. We trapped deer during winter months on two study areas (Northern Forest and Eastern Farmland) in Wisconsin from 2010 to 2013. Deer were tested for exposure to six serovars of Leptospira interrogans (grippotyphosa, icterohaemorrhagiae, canicola, bratislava, pomona, and hardjo), bovine viral diarrhea virus (BVDV-1 and BVDV-2), infectious bovine rhinotracheitis virus (IBR), and parainfluenza 3 virus (PI3). We used logistic regression to model potential intrinsic (e.g., age, sex) and extrinsic (e.g., land type, study site, year, exposure to multiple pathogens) variables we considered biologically meaningful to exposure of deer to livestock pathogens. Deer sampled in 2010-2011 did not demonstrate exposure to BVDV, so we did not test for BVDV in subsequent years. Deer had evidence of exposure to PI3 (24.7%), IBR (7.9%), Leptospira interrogans serovar pomona (11.7%), L. i. bratislava (1.0%), L. i. grippotyphosa (2.5%) and L. i. hardjo (0.3%). Deer did not demonstrate exposure to L. interrogans serovars canicola and icterohaemorrhagiae. For PI3, we found that capture site and year influenced exposure. Fawns (n = 119) were not exposed to L. i. pomona, but land type was an important predictor of exposure to L. i. pomona for older deer. Our results serve as baseline exposure levels of Wisconsin white-tailed deer to livestock pathogens, and helped to identify important factors that explain deer exposure to livestock pathogens.

Evaluation of Trapper-Collected Nobuto Filter-Paper Blood Samples for Distemper and Parvovirus Antibody Detection in Coyotes (Canis latrans) and Raccoons (Procyon lotor).

Blood samples are often collected from free-ranging wildlife for antibody detection. However, filter-paper (FP) strips are more cost efficient and easy to collect and store. We evaluated trapper-collected FP strips and body-cavity blood for canine distemper (CDV) and parvovirus (CPV-2) antibody detection in raccoons (Procyon lotor) and coyotes (Canis latrans). From 2008 to 2010, licensed trappers near Madison and Milwaukee, Wisconsin, US collected paired samples from harvested animals. Canine distemper antibodies were detected using virus neutralization and parvovirus antibodies were detected using hemagglutination inhibition. Titers ≥ 1:32 for CDV and ≥ 1:25 for CPV-2 were considered evidence of exposure. Using Cohen's kappa test of agreement, FP strip titers agreed with sera for CDV in coyotes (n = 28, K = 0.772) and raccoons (n = 29, K = 0.858) and for CPV-2 in coyotes (n = 40, K = 0.775) and raccoons (n = 70, K = 0.646). However, raccoons determined to be exposed to CPV-2 from sera were unexposed by FP strips in 35% of the samples. Titer results may be affected by quality and volume of blood samples, interval between collection and processing, small sample sizes, and diagnostic testing procedures. Filter-paper strips can be useful for detecting CDV and CPV-2 exposure in coyotes and raccoons with correct field sample collection and appropriate diagnostic testing procedures.

Influence of ecologic factors on prevalence of meningeal worm (Parelaphostrongylus tenuis) infection in South Dakota, USA.

The meningeal worm (Parelaphostrongylus tenuis) is a nematode parasite that commonly infects white-tailed deer (Odocoileus virginianus; WTD) throughout the deciduous forest biome and deciduous-coniferous ecotone of eastern and central North America; the species is not known to occur west of the grassland biome of central North America. We used county-specific prevalence data to evaluate potential effects of landscape and climatologic factors on the spatial distribution of meningeal worm infection in South Dakota, US. Probability of infection increased 4-fold between eastern and western South Dakota and 1.3-fold for each 1-cm increase in summer precipitation. Sixty-three percent of WTD had only a single worm in the cranium. Expansion of meningeal worm infection across western South Dakota may be inherently low due to the combined effects of arid climate and potential attributes of the Missouri River that limit regional movements by infected WTD. Use of landscape genetic analyses to identify potential relationships between landscape features and population genetic structure of infected deer and parasites may contribute to a greater understanding of regional heterogeneity in meningeal worm infection rates across South Dakota, particularly in counties adjacent to the Missouri River. Future research evaluating heterogeneity in prevalence and intensity of infection between fawn and yearling deer, and the potential role of yearling male deer as dispersal agents of meningeal worms across the Missouri River, also is warranted.

Prevalence and intensity of nematode parasites in Wisconsin ermine.

In the midwestern United States, ermine ( Mustela erminea ) are economically important because they are legally harvested for pelts. Information on parasites of ermine is lacking, and the effects that nematode parasites have on body condition of ermine hosts are unknown. We identified Skrjabingylus nasicola and Filaroides martis in ermine trapped from 2007 to 2013 from 6 counties in Wisconsin. Small mammals, commonly consumed by ermine, serve as paratenic hosts for both parasites. Our goal was to identify how age and sex of ermine, along with year, influence nematode parasitism. We also investigated how infection affected body condition for male and female ermine using body mass standardized by length as an index of body condition. We commonly found S. nasicola and F. martis in male and female ermine, but both prevalence and intensity of infection were higher for males. Relative to juveniles (<1 yr), adult (>1 yr) male ermine did not exhibit significantly higher intensity or prevalence of either parasite. We found that body condition was not compromised by infection for either sex, and intensity of S. nasicola and prevalence of F. martis were highest during the 2010-2011 trapping season. Of the 6 yr studied, precipitation was highest during the summer before the 2010-2011 season, and increased precipitation can cause increases in populations of gastropod intermediate hosts. We think that several distinct natural history components, namely, mating structure, diet, and metabolic rate, influence nematode parasitism in ermine.

Influence of environmental variables on Baylisascaris procyonis infection in raccoons.

Baylisascaris procyonis is a zoonotic nematode commonly found in raccoons (Procyon lotor). Human-altered landscapes can support dense populations of raccoons, increasing the potential for interaction between humans and these animals. We used raccoon feces provided by licensed fur trappers to investigate environmental variables that influence prevalence of B. procyonis at 2 sites in Wisconsin. Trappers submitted raccoon feces to us, along with information on sex, age (juvenile and adult), and approximate trap location for each animal. We used zinc sulfate (1.18 specific gravity) flotation to detect B. procyonis eggs in approximately 1 g of fecal matter from each host. We used ArcView software to determine the distance of each trap location to an urban area as determined by the U.S. Census Bureau in 2000. We compared the habitat components in buffered home ranges (0.805 km around trap locations) of infected animals with those from uninfected animals using Mann-Whitney U-tests (P < 0.05). Variables investigated were human population, road density, housing units per census block, and land cover, including area of agriculture, forest, developed, shrubland, water, and grassland. We positively identified eggs in 64.9% of the animals sampled. Raccoons infected with B. procyonis had significantly larger area of agricultural habitats and significantly smaller areas of forested habitats in buffered home ranges than uninfected individuals. We found that raccoons near Madison and Milwaukee, Wisconsin, are commonly infected with B. procyonis , indicating that public education regarding protection from disease is warranted.

Parasite infections in nestling red-shouldered hawks (Buteo lineatus) in northeast Wisconsin.

Red-shouldered hawks (Buteo lineatus) are threatened in Wisconsin and long-term data suggest that nest productivity is low in the state for unknown reasons. Our objective was to determine whether red-shouldered hawks in northeast Wisconsin were infected with parasites that could contribute to low nest productivity. We examined nestlings for the presence of Trichomonas gallinae, Protocalliphora avium, and blood parasites in June 2006 and 2007. We did not detect T. gallinae in throat swabs taken from 24 nestlings in 2007. Ear canals of nestlings were parasitized by P. avium larvae in 10 of 11 (91%) nests and in 22 of 24 (92%) nestlings. Larvae were found in higher intensity in 1 ear relative to the other. Leucocytozoon toddi was present in 90.5% (38/42) of the nestlings. At least 1 bird in each nest was infected. Intensity of L. toddi averaged 48.6 +/- 58.3 infected cells per 2,000 erythrocytes (2.4 +/- 2.9%). No other blood parasites were identified.

Determining prevalence of bluetongue and epizootic hemorrhagic disease viruses in mule deer in Arizona (USA) using whole blood dried on paper strips compared to serum analyses.

We investigated the feasibility of using whole blood dried on paper strips as a means to collect antibody prevalence data for the epizootic hemorrhagic disease viruses (EHDV) and bluetongue viruses (BTV) from hunter-harvested male mule deer (Odocoileus hemionus) in October 2002 from Arizona, USA. We compared antibody prevalence estimates in mule deer from paired paper strip and serum samples. Prevalence data obtained from elution of dried blood on paper strips proved to be consistent with results from serum in 94% of the samples tested. The paper strip method allows easy collection of blood from dead animals, with a smaller amount of blood being needed for analyses. Also, samples do not need to be refrigerated before analyses. We also used serum samples to determine hemorrhagic disease (HD) serotype exposure status of mule deer harvested from 4 distinct areas in Arizona. Antibodies to BTV and EHDV were identified in 3 of the 4 areas, with positive results to EHDV-1, EHDV-2, BTV-10, and BTV-11 being most common. Many animals did not have antibodies against the BTV serotypes. Exposure varied geographically and potentially with elevation. Hemorrhagic disease viruses commonly infect Arizona mule deer, except on the Kaibab Plateau in northern Arizona.

Serologic survey for pathogens potentially affecting pronghorn (Antilocapra americana) fawn recruitment in Arizona, USA.

During the 1990s, pronghorn (Antilocapra americana) populations declined in Arizona, USA. To investigate potential causes of decline, we collected blood samples from hunter-harvested male pronghorn from 2001 to 2003 on four Arizona sites. Sera were tested for antibody to parainfluenza virus type 3 (PI3), bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, bovine respiratory syncytial virus, epizootic hemorrhagic disease virus (EHDV), bluetongue virus (BTV), and Chlamydia psittaci. Antibody against PI3 was found in 33% of the samples, whereas antibody against BTV/EHDV was found in 77%. Antibodies to other pathogens were found at low prevalence rates. Although pronghorn decline in Arizona is probably not directly related to disease, potential reproductive effects of BTV/EHDV and PI3 infection on pronghorn in Arizona merit further study.

Evidence of tick-borne organisms in mule deer (Odocoileus hemionus) from the western United States.

Free-ranging mule deer (MD; Odocoileus hemionus) from Arizona and California were tested for evidence of infection with several tick-borne pathogens, including species of Ehrlichia, Anaplasma, Babesia, and Borrelia. Of 125 mule deer tested from Arizona, 29 (23%) and 11 (9%) had antibodies reactive to E. chaffeensis and A. phagocytophilum by indirect immunofluorescent antibody testing, respectively; none of the six MD tested from California were seropositive. Using a commercial competitive ELISA kit, antibodies reactive to Anaplasma spp. were detected in 19 (15%) MD from Arizona and four of six (67%) MD from California. Polymerase chain reaction (PCR) testing for tick-borne pathogens was conducted on blood samples from 29 MD from Arizona and 11 MD from California. Twenty-two of 29 (75.9%) MD from Arizona had PCR evidence of infection with at least one tick-borne pathogen. We detected an Anaplasma sp. in 19 of 29 (65.5%) MD and a Babesia sp. in 10 of 29 (34%) MD. Sequencing of these amplicons indicated that the Anaplasma sp. was the same that had previously been detected in MD from California and the Babesia sp. was similar to one previously detected in a reindeer (Rangifer tarandus tarandus) from California. All of the California MD had evidence of infection with a tick-borne pathogen. Two different species of Anaplasma spp. were detected in MD from California, eight of of 11 MD were infected with an Anaplasma sp., and three of 11 MD were infected with A. ovis. This is the first report of a mule deer naturally infected with A. ovis. Ten of 11 MD from California were infected with a Babesia-like organism previously associated with human disease, and a single MD was PCR positive for Borrelia coriaceae, which has been associated with epizootic bovine abortion. Together, these data suggest that MD in northern Arizona and eastern California are exposed to several pathogens of human and veterinary importance.

Strongyloides robustus and the northern sympatric populations of northern (Glaucomys sabrinus) and southern (G. volans) flying squirrels.

Within North America, northern (Glaucomys sabrinus) and southern (Glaucomys volans) flying squirrels occupy distinct ranges with limited overlap. Sympatry in northern latitudes coincides with northern hardwood vegetation from Minnesota to New England. Strongyloides robustus is an intestinal parasite that infects both species but appears to be deleterious only to northern flying squirrels. As a result, S. robustus could be a critical determinant of flying squirrel population characteristics in at least some areas of sympatry. However, cold weather could potentially limit the distribution of S. robustus in northern climates. Therefore, we assessed fecal samples from both flying squirrel species to determine the presence of the nematode in Wisconsin. Strongyloides robustus was found in 12 flying squirrel scat samples and infected 52% of southern flying squirrels and 11% of northern flying squirrels. Prevalence of S. robustus infection for northern flying squirrels was substantially lower than previously reported from more southern regions. This is the northernmost documentation of S. robustus in flying squirrels and the first documentation of S. robustus parasitizing flying squirrels in Wisconsin.

Epizootiology of hemorrhagic disease in mule deer in central Arizona.

We report two male mule deer (Odocoileus hemionus) mortalities that occurred near Prescott, Arizona (USA) in September 2001. Necropsy lesions were compatible with those described for hemorrhagic disease (HD). Bluetongue (BTV) and epizootic hemorrhagic disease (EHDV) viral RNA were identified in tissues from one deer and EHDV viral RNA was identified in the other via reverse transcription-polymerase chain reaction (RT-PCR). Virus isolation attempts were unsuccessful in both cases. To determine geographic range and viral serotypes involved, we collected blood samples from healthy hunter-harvested male mule deer in three game management units near Prescott in October 2001. Forty-one blood samples were collected and antibodies against EHDV and/or BTV were found in 21/41 (51%) samples as determined by agar immunodiffusion. Serum neutralization test results suggest that multiple EHDV and BTV serotypes are present in Arizona.