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.

Caridean Shrimps Associated with the Slimy Sea Plume (Pseudopterogorigia americana) in Midsummer at Guana Island, British Virgin Islands, West Indies

Fifty-one slimy sea plumes (Pseudopterogorgia americana Gmelin, 1791) were sampled for caridean shrimps at Guana Island, British Virgin Islands, during one week in July 1992. Sam- pling depth ranged from 3-22 m. Nine species were collected: Hippolyte nicholsoni Chace, 1972; Latreutes sp.; Neopontonides chacei Heard, 1986; Perclimenes cf. patae Heard and Spotte, 1991; Periclimenes cf. pauper Holthuis, 1951; Periclimenes sp.; Pseudocoutierea antillensis Chace, 1972; Tozeuma cf. cornutum Milne Edwards, 1881; and Trachycaris rugosa (Bate, 1888). A total of 1,418 specimens (including fragments) was obtained. The number of shrimp species per gorgonian ranged from 1-5; one gorgonian harbored 156 shrimps. The two predominant species, N. chacei and H. nicholsoni, occupy different mean depths (12.6 and 8.2 m, respectively). Sexual dimorphism assessed with Mann-Whitney U-tests was not apparent in the specimens of N. chacei (P > 0.05), but females of H. nicholsoni were significantly larger than males (P < 0.001). Minimum carapace length (CL, the tip of the rostrum to the posterior dorsal margin of the carapace) at which male N. chacei acquire a single appendix masculina spine is 1.25 mm; male H. nicholsoni can acquire a single spine at 0.9 mm CL. Histological sections of male N. chacei showed that shrimp with 0 or 1 spine are least likely to be mature. Female N. chacei can become ovigerous at 1.9 mm CL and female H. nicholsoni at 1.2 mm CL. The taxonomic status of 5 of the 9 species collected is uncertain.

Virus-Associated Tumours of the Harderian Gland in Laboratory-Reared Voles, Clethrionomys rutilis (Pallas)

Tumors of the Harderian gland occurred frequently in red-backed voles, Clethrionomys rutilus (Pallas), in a captive colony maintained since 1957. Although the tumors became obvious only in animals about a year old and older, early stage changes were histologically evident in glands from younger, clinically normal voles. The tumors ranged in type from poorly differentiated adenoma to adenocarcinoma, with invasion of adjacent tissues and characteristic deformation of the zygomata. Electron micrographs disclosed viral particles in the cytoplasm of glands from clinically normal animals as well as in glands showing malignant changes. The tumor was not observed in arvicoline rodents of other species maintained in the same rooms with the red-backed voles.

An Investigation of Wild Bee Diversity and Abundance in Plots Managed by The Nature Conservancy in South-Central Nebraska and of Beneficial Arthropods Associated with Native Nebraska Flora

Insect pollination is an essential ecosystem service, and bees are the principal pollinators of wild and cultivated plants. Habitat management and enhancement are a proven way to encourage wild bee populations, providing them with food and nesting resources. I examined bee diversity and abundance in plots managed by The Nature Conservancy near Wood River, NE. The plots were seeded with 2 seed mixes at 2 seeding rates: high diversity mix at the recommended rate, high diversity mix double the recommended rate, Natural Resources Conservation Service (NRCS) conservation planting (CP) 25 mix at one-half the recommended rate, and NRCS CP25 mix at the recommended rate. I measured wild bee abundance and diversity, and established a database of wild bees associated with the plots. I also compared genus richness and abundance among the plots using and aerial net and blue vane traps to collect bees. Significant differences were not observed in genus richness and diversity among the plots; however, plot size and the ability of blue vane traps to draw bees from a long distance may have influenced my results. In 2008, 15 genera and 95 individual bees were collected using an aerial net and in 2009, 32 genera and 6,103 individual bees were collected using blue vane traps. I also studied the beneficial insects associated with native Nebraska flora. Seventeen species of native, perennial flora were established in 3 separate plots located in eastern Nebraska. I transplanted four plants of each species in randomized 0.61 m x 0.61 m squares of a 3.05 m x 9.14 m plot. Arthropods were sampled using a modified leaf blower/vacuum. Insects and other arthropods were identified to family and organized into groups of predators, parasites, pollinators, herbivores, and miscellaneous. Associations between plant species and families of beneficial arthropods (predators, parasites, and pollinators) were made. Pycnanthemum flexuosum Walter attracted significantly more beneficial arthropod families than 7 other species of plants tested. Dalea purpurea Vent and Liatris punctata Hook also attracted significantly fewer beneficial arthropod families than 4 other species of plants tested. In total, 31 predator, 11 parasitic, 4 pollinator, 31 herbivore, and 10 miscellaneous families of arthropods were recorded.

Animal-side serologic assay for rapid detection of Mycobacterium Bovis infection in multiple species of free-ranging wildlife

Numerous species of mammals are susceptible to Mycobacterium bovis, the causative agent of bovine tuberculosis (TB). Several wildlife hosts have emerged as reservoirs of M. bovis infection for domestic livestock in different countries. In the present study, blood samples were collected from Eurasian badgers (n = 1532), white-tailed deer (n = 463), brushtail possums (n = 129), and wild boar (n = 177) for evaluation of antibody responses to M. bovis infection by a lateral-flow rapid test (RT) and multiantigen print immunoassay (MAPIA). Magnitude of the antibody responses and antigen recognition patterns varied among the animals as determined by MAPIA; however, MPB83 was the most commonly recognized antigen for each host studied. Other seroreactive antigens included ESAT-6, CFP10, and MPB70. The agreement of the RT with culture results varied from 74% for possums to 81% for badgers to 90% for wild boar to 97% for white-tailed deer. Small numbers of wild boar and deer exposed to M. avium infection or paratuberculosis, respectively, did not cross-react in the RT, supporting the high specificity of the assay. In deer, whole blood samples reacted similarly to corresponding serum specimens (97% concordance), demonstrating the potential for field application. As previously demonstrated for badgers and deer, antibody responses to M. bovis infection in wild boar were positively associated with advanced disease. Together, these findings suggest that a rapid TB assay such as the RT may provide a useful screening tool for certain wildlife species that may be implicated in the maintenance and transmission of M. bovis infection to domestic livestock.

Estimating the extent of spatial association of Mycobacterium bovis infection in badgers in Ireland

Mycobacterium bovis infects the wildlife species badgers Meles meles who are linked with the spread of the associated disease tuberculosis (TB) in cattle. Control of livestock infections depends in part on the spatial and social structure of the wildlife host. Here we describe spatial association of M. bovis infection in a badger population using data from the first year of the Four Area Project in Ireland. Using second-order intensity functions, we show there is strong evidence of clustering of TB cases in each the four areas, i.e. a global tendency for infected cases to occur near other infected cases. Using estimated intensity functions, we identify locations where particular strains of TB cluster. Generalized linear geostatistical models are used to assess the practical range at which spatial correlation occurs and is found to exceed 6 in all areas. The study is of relevance concerning the scale of localized badger culling in the control of the disease in cattle.

Aerosol Exposure of White-Tailed Deer (Odocoileus virginianus) to Mycobacterium Bovis

Tuberculosis due to Mycobacterium bovis affects both captive and free-ranging Cervidae in the United States. Various animal models have been developed to study tuberculosis of both humans and animals. Generally, tuberculosis is transmitted by aerosol and oral routes. Models of aerosol exposure of large animals to M. bovis are uncommon. In order to develop a reliable method of aerosol exposure of white-tailed deer (Odocoileus virginianus) to M. bovis, 12 healthy white-tailed deer, aged 8–10 mo, were infected by aerosol exposure to 2x105 to 1x106 colony forming units (CFU) (high dose, n=4) of M. bovis or 6x102 to 1.6 x 103 CFU (low dose, n=8) of M. bovis. Tuberculous lesions were more widely disseminated in deer receiving the high dose, while lesions in deer receiving the low dose were more focused on the lungs and associated lymph nodes (tracheobronchial and mediastinal). Aerosol delivery of M. bovis to white-tailed deer results in a reliable manner of experimental infection that may be useful for studies of disease pathogenesis, immune response, mycobacterial shedding, and vaccine efficacy.

Antibody Responses in Reindeer (Rangifer tarandus) Infected with Mycobacterium bovis

Despite having a very low incidence of disease, reindeer (Rangifer tarandus) are subject to tuberculosis (TB) testing requirements for interstate shipment and herd accreditation in the United States. Improved TB tests are desperately needed, as many reindeer are falsely classified as reactors by current testing procedures. Sera collected sequentially from 11 (experimentally) Mycobacterium bovis-infected reindeer and 4 noninfected reindeer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for antibody specific to M. bovis antigens. Specific antibody was detected as early as 4 weeks after challenge with M. bovis. By MAPIA, sera were tested with 12 native and recombinant antigens, which were used to coat nitrocellulose. All M. bovis-infected reindeer developed responses to MPB83 and a fusion protein, Acr1/MPB83, and 9/11 had responses to MPB70. Other antigens less commonly recognized included MPB59, ESAT-6, and CFP10. Administration of purified protein derivatives for skin testing boosted serum antibody responses, as detected by each of the assays. Of the noninfected reindeer, 2/4 had responses that were detectable immediately following skin testing, which correlated with pathological findings (i.e., presence of granulomatous lesions yet the absence of acid-fast bacteria). The levels of specific antibody produced by infected reindeer appeared to be associated with disease progression but not with cell-mediated immunity. These findings indicate that M. bovis infection of reindeer elicits an antibody response to multiple antigens that can be boosted by skin testing. Serological tests using carefully selected specific antigens have potential for early detection of infections in reindeer.

Paraophioidina scolecoides n. sp., a New Aseptate Gregarine from Cultured Pacific White Shrimp Penaeus vannamei

The aseptate gregarine Paraophloidina scolecoides n. sp. (Eugregarinorida: Lecudinidae) heavily infected the midgut of cultured larval and postlarval specimens of Penaeus vannamei from a commercial 'seed-production' facility in Texas, USA. It is morphologically similar to P. korotneffi and P. vibiliae, but it can be distinguished from them and from other members of the genus by having gamonts associated exclusively by lateral syzygy. Shrimp acquired the infection at the facility; nauplii did not show any evidence of infection, but protozoea, mysis, and postlarval shrimp had a prevalence and intensity of infection ranging from 56 to 80 % and 10 to >50 parasites, respectively. Infected shrimp removed from the facility to aquaria at another location lost their gamont infection within 7 days When voided from the gut, the gregarine disintegrated in seawater. Results suggest that P. vannamei is an accidental host, although a survey of representative members of the invertebrate fauna from the environment associated with the facility failed to discover other hosts. No link was established between infection and either the broodstock or the water or detritus from the nursery or broodstock tanks.

Relationship Between BP (Baculovirus penaei) Energy Reserves in Larval and Postlarval Pacific White Shrimp Penaeus vannamei

The relationship between energy reserves of the penaeid shrimp Penaeus vannamei and Baculovirus penaei, or BP, were investigated in a series of experiments using mysis stage or early postlarval shrimp. Pre-exposure and post-exposure levels of protein and triacylgycerol (TAG) were determined. The effect of pre-exposure protein and TAG levels on susceptibility to BP infections was also investigated by starving a group of shrimp immediately prior to BP exposure. There was no consistent relationship between either pre-exposure or post-exposure protein levels and the percent of shrimp developing patent BP infections. There was, however, a significant positive correlation between TAG levels immediately prior to viral exposure and prevalence of infection 72 h later. Experimental reduction of TAG reserves prior to BP exposure delayed the development of a patent infection. In some, but not all, experiments there was a significant reduction in TAG levels of infected compared with uninfected shrimp 72 h post-exposure. The effect of patent BP infections on host TAG levels was subordinate to fluctuations in TAG content associated with the ontogeny of the hepatopancreas. Results of this study support histological observations that shrimp lipid levels can be altered by baculovirus infections. Furthermore, high levels of energy reserves in the form of TAG are associated with increased susceptibility to BP infection in larval and postlarval shrimp.

Book Review of Cetacean Societies: Field Studies Of Whales And Dolphins Edited by I. Mann, R. C. Connor, P. L. Tyack & H. Whitehead

Studying the sociobiology and behavioral ecology of cetaceans is particularly challenging due in large part to the aquatic environment in which they live. Nevertheless, many of the obstacles traditionally associated with data gathering on tree-ranging whales, dolphins and porpoises are rapidly being overcome, and are now far less formidable. During the past several decades, marine mammal scientists equipped with innovative research methods and new technologies have taken field-based behavioral studies to a new level of sophistication. In some cases, as is true for bottlenose dolphins, killer whales, sperm whales and humpback whales, modern research paradigms in the marine environment are comparable to present-day studies of terrestrial mammal social systems. Cetacean Society stands testament to the relatively recent advances in marine mammal science, and to those scientists, past and present, whose diligence has been instrumental in shaping the discipline.

Book Review of Entomology at the Land Grant University: Perspectives from the Texas A&M University Department Centenary Edited by Kevin M. Heinz, Raymond Frisbie, and Carlos Bogran

This book offers the texts of twenty-four presentations given in a series of mini symposia organized to commemorate the centennial of Texas A&M's Department of Entomology. Although not stated explicitly by the editors, the central theme for these symposia seems to be the contributions of the discipline of entomology to modern society. The presentations cover a wide range of topics dealing with the importance of insects in both natural ecosystems and agricultural systems; insects as models for scientific research; challenges associated with effective management of those species that are pests in systems designed for food and fiber production; and the role of entomology departments within academic institutions, particularly land-grant universities. The list of contributors to these symposia is impressive, as are the universities and other institutions they represent across the United States as well as in Europe and Africa.