Risky business in Georgia's wild birds: contact rates between wild birds and backyard chickens is influenced by supplemental feed.

Backyard chickens are increasingly popular, and their husbandry varies widely. How backyard chickens are housed may influence the accessibility of chicken feed and water to wild birds, and thus, the contact rates between both groups. Increased contacts have implications for pathogen transmission; for instance, Newcastle disease virus or avian influenza virus may be transmitted to and from backyard chickens from contaminated water or feed. Given this potentially increased pathogen risk to wild birds and backyard chickens, we examined which wild bird species are likely to encounter backyard chickens and their resources. We performed a supplemental feeding experiment followed by observations at three sites associated with backyard chickens in North Georgia, USA. At each site, we identified the species of wild birds that: (a) shared habitat with the chickens, (b) had a higher frequency of detection relative to other species and (c) encountered the coops. We identified 14 wild bird species that entered the coops to consume supplemental feed and were considered high-risk for pathogen transmission. Our results provide evidence that contact between wild birds and backyard chickens is frequent and more common than previously believed, which has crucial epidemiological implications for wildlife managers and backyard chicken owners.

Detection of Bacillus anthracis in animal tissues using InBios active anthrax detect rapid test lateral flow immunoassay.

The Active Anthrax Detect (AAD) Rapid Test lateral flow immunoassay is a point-of-care assay that was under investigational use for detecting Bacillus anthracis capsular polypeptide (polyglutamic acid) in human blood, serum and plasma. Small sample volumes, rapid results and no refrigeration required allow for easy use in either the field or laboratory. Although the test was developed for use in suspect cases of human inhalation anthrax, its features also make it a potentially powerful tool for testing suspect animal cases. We tested animal tissue samples that were confirmed or ruled out for B. anthracis. The AAD Rapid Tests were also deployed in the field, testing animal carcasses during an anthrax outbreak in hippopotami (Hippopotamus amphibius) and Cape buffalo (Syncerus caffer) in Namibia. Evaluation of all samples showed a specificity of 82% and sensitivity of 98%. However, when the assay was used on specimens from only fresh carcasses (dead for <24 h), the specificity increased to 96%. The AAD Rapid Test is a rapid and simple screening assay, but confirmatory testing needs to be done, especially when the age of the sample (days animal has been deceased) is unknown. SIGNIFICANCE AND IMPACT OF THE STUDY: In countries where anthrax is endemic, many human outbreaks are often caused by epizootics. Earlier detection of infected animals may allow for identification of exposed people, early implementation of prevention and control methods, and ultimately lessen the number of people and animals affected. Detection of Bacillus anthracis in animal tissues using a simple, rapid and field-deployable method would allow for faster outbreak response. We evaluated a simple sample collection and processing method for use with the Active Anthrax Detect Rapid Test lateral flow immunoassay to screen dead animals for anthrax.

Besnoitia neotomofelis n. sp. (Protozoa: Apicomplexa) from the southern plains woodrat ( Neotoma micropus).

Certain species of the protozoan genus Besnoitia cause clinical disease in livestock and wildlife. In the present paper a new species, Besnoitia neotomofelis is described from the southern planes woodrat (Neotoma micropus). The parasite was detected by bioassay of woodrat tissues in outbred Swiss Webster mice in an attempt to isolate Toxoplasma gondii. Initially, the organism was misdiagnosed as T. gondii because it was highly pathogenic for mice and its tachyzoites resembled T. gondii tachyzoites. Further studies revealed that it differed structurally and biologically from T. gondii. Tachyzoites were successfully cultivated and maintained in vitro in bovine monocytes and African green monkey kidney cells, and in vivo in mice. Non-dividing, uninucleate tachyzoites were approximately 1 x 5 µm in size. Longitudinally-cut bradyzoites in tissue sections measured 1.5-1.6 x 7.7-9.3 µm. Tissue cysts were microscopic, up to 210 µm long, and were infective orally to mice. Cats fed tissue cysts shed unsporulated 13 x 14 µm sized oocysts. All mice inoculated with B. neotomofelis died of acute besnoitiosis, irrespective of the dose, and Norwegian rats became infected but remained asymptomatic. Entero-epithelial stages (schizonts, gamonts) were found in cats fed tissue cysts. Large (up to 40 x 50 µm) first-generation schizonts developed in the lamina propria of the small intestine of cats. A second generation of small sized (8 µm) schizonts containing 4-8 merozoites was detected in enterocytes of the small intestine. Gamonts and oocysts were seen in goblet cells of the small intestinal epithelium. Tachyzoites were present in mesenteric lymph nodes of cats. Phylogenetic analysis indicated that B. neotomofelis was related to other Besnoitia species from rodents, rabbits, and opossums. Besnoitia neotomofelis is distinct from the 3 other species of Besnoitia, B. wallacei, B. darlingi and B. oryctofelisi that utilize cats as a definitive host.

Ecological havoc, the rise of white-tailed deer, and the emergence of Amblyomma americanum-associated zoonoses in the United States.

Two infectious diseases, and one presumably infectious disease, each vectored by or associated with the bite of the lone star tick (Amblyomma americanum), were identified and characterized by clinicians and scientists in the United States during the 1980s and 1990s. These three conditions-human monocytic (or monocytotropic) ehrlichiosis (HME), Ehrlichia ewingii ehrlichiosis, and southern tick-associated rash illness (STARI)-undoubtedly existed in the United States prior to this time. However, the near-simultaneous recognition of these diseases is remarkable and suggests the involvement of a unifying process that thrust multiple pathogens into the sphere of human recognition. Previous works by other investigators have emphasized the pivotal role of white-tailed deer (Odocoileus virginianus) in the emergence of Lyme disease, human babesiosis, and human granulocytic anaplasmosis. Because whitetails serve as a keystone host for all stages of lone star ticks, and an important reservoir host for Ehrlichia chaffeensis, E. ewingii, and Borrelia lonestari, the near-exponential growth of white-tailed deer populations that occurred in the eastern United States during the twentieth century is likely to have dramatically affected the frequency and distribution of A. americanum-associated zoonoses. This chapter describes the natural histories of the pathogens definitively or putatively associated with HME, E. ewingii ehrlichiosis, and STARI; the role of white-tailed deer as hosts to lone star ticks and the agents of these diseases; and the cascade of ecologic disturbances to the landscape of the United States that have occurred during the last 200 years that provided critical leverage in the proliferation of white-tailed deer, and ultimately resulted in the emergence of these diseases in human populations.

Raccoon roundworm (Baylisascaris procyonis) as an occupational hazard: 1. Knowledge of B. procyonis and attitudes towards it and other zoonoses among wildlife rehabilitators.

Wildlife rehabilitators are at risk of zoonotic diseases because they often have prolonged contact with many species of wildlife and their bodily fluids. Raccoon roundworm (Baylisascaris procyonis) is a common zoonotic parasite of raccoons that has the potential to cause severe or fatal neurologic disease in a broad variety of hosts if the eggs within raccoon faeces are ingested. We administered an online survey to wildlife rehabilitators to assess their knowledge regarding aspects of transmission, biology and disease caused by B. procyonis, and also to evaluate attitudes towards wildlife diseases and B. procyonis as an occupational hazard. Knowledge was assessed using multiple choice and true-false questions; attitudes were measured using Likert-type items. A total of 659 complete or near-complete responses (missing fewer than three knowledge or attitudes items and/or non-response to some demographic fields) were collected. The median knowledge score was 7/14 questions correct (range: 0-14 correct). Generally, individuals with higher levels of education and rehabilitation experience, veterinary professionals and those who are members of professional wildlife rehabilitation groups scored above the median significantly more often (p < .01). Significantly more rehabilitators who were located in the south-east and those with part-time or infrequent commitments scored below the median overall knowledge score. There was general agreement that B. procyonis is a health risk of rehabilitators and that measures should be taken to control transmission to people and animals. Some factors explaining differences in attitudes include setting of rehabilitation (home versus animal care facility), veterinary profession, region, membership in a wildlife rehabilitation group and rehabilitation of raccoons. Findings emphasize the importance of awareness and mentorship to inform rehabilitators on the potential risks of B. procyonis and other potential zoonoses within captive wildlife settings, and the important role of professional wildlife rehabilitator groups in disseminating educational materials.

Raccoon roundworm (Baylisascaris procyonis) as an occupational hazard: 2. Use of personal protective equipment and infection control practices among raccoon rehabilitators.

Baylisascaris procyonis, the raccoon roundworm, is a zoonotic ascarid of importance to human and animal health. Wildlife rehabilitators who care for raccoons may be at an increased risk for exposure to the parasite, especially if proper precautions are not taken. In a wider effort to evaluate awareness regarding B. procyonis in the wildlife rehabilitation community, an online survey (38-39 questions) including questions about B. procyonis knowledge and attitudes was developed and administered to wildlife rehabilitators. To assess precautions taken among raccoon rehabilitators, participants who rehabilitated raccoons (n = 447) answered additional questions about use of personal protective equipment (PPE) and infection control practices (ICPs). Reported use of gloves was variable, but hand hygiene was generally consistent. Masks and gowns were seldom used. Part-time or infrequent volunteers and rehabilitators located in the Central, Midwest and Southeast were significantly less likely to report consistent use of PPE. A total knowledge score from the survey was used to predict the likelihood of reporting the use of particular ICPs/PPE. Knowledge score had a highly significant but small effect on the likelihood of prophylactic use of anthelmintics, anthelmintics use for B. procyonis specifically, cleaning appropriately, and using species-dedicated housing. Risk factor analysis was performed on data from a prior serologic survey to evaluate factors associated with exposure to B. procyonis and inconsistent handwashing after contact with live raccoons and their faeces; practising rehabilitation in B. procyonis hyperendemic regions and practising rehabilitation in the western region were significant risk factors for being seropositive. These data further demonstrate that correct PPE/ICPs are critical in mitigating the risk of B. procyonis exposure among raccoon rehabilitators and among other captive species.

Schistosomes and Microfilarial Parasites in Magellanic Penguins.

The Magellanic Penguin ( Spheniscus magellanicus) is native to Argentina, Chile, and the Falkland/Malvinas Islands, and is a regular winter migrant in Uruguayan and Brazilian coastal waters. The species is known to be susceptible to a variety of gastrointestinal nematodes, cestodes, trematodes, and acanthocephalans, as well as renal trematodes and pulmonary nematodes. Schistosomes (Platyhelminthes, Trematoda, Schistosomatidae) and microfilariae (Nematoda, Secernentea, Onchocercidae) were histologically identified in Magellanic Penguins ( Spheniscus magellanicus) that died while under care at rehabilitation centers in southern Brazil. Phylogenetic analysis of the COI gene, ITS-1 region, 5.8S rRNA gene, ITS-2 region, and 28S rRNA gene sequences of the schistosome revealed that it is closely related to, but distinct from, a schistosome reported from the African Penguin ( Spheniscus demersus). The schistosomes from Magellanic and African Penguins were grouped with Gigantobilharzia huronensis, Gigantobilharzia melanoidis, and Dendritobilharzia pulvurenta; however, the lack of a clearly monophyletic origin precludes determining their genus. The incidental discovery of novel parasites during a study that did not specifically aim to investigate the occurrence of helminths underscores the value of histopathological examination as an exploratory diagnostic approach.

Habitat factors influencing distributions of Anaplasma phagocytophilum and Ehrlichia chaffeensis in the Mississippi Alluvial Valley.

Human monocytotropic ehrlichiosis (HME), caused by the bacterium Ehrlichia chaffeensis, and human granulocytic anaplasmosis (HGA), caused by the bacterium Anaplasma phagocytophilum, are two emerging tick-borne zoonoses of concern. Factors influencing geographic distributions of these pathogens are not fully understood, especially at varying spatial extents (regional versus landscape) and resolutions (counties versus smaller land units). We used logistic regression to compare influences of physical environment, land cover composition, and landscape heterogeneity on distributions of A. phagocytophilum and E. chaffeensis at multiple spatial extents. Pathogen presence or absence was determined from white-tailed deer (Odocoileus virginianus) serum samples collected from 1981 to 2005. Ecological predictor variables were derived from spatial datasets that represented deer density, elevation, land cover, normalized difference vegetation index (NDVI), hydrology, and soil moisture. We used three strategies (a priori, exploratory, and spatial extent) to develop models. Best fitting models were applied within a geographic information system to create predictive probability surfaces for each bacterium. Ecological predictor variables generally resulted in better fitting models for E. chaffeensis than A. phagocytophilum (90.5% and 68% sensitivity, respectively), possibly as a result of differences in the natural histories of tick vectors. Although alternative model development strategies produced different models, in all cases bacteria presence or absence was affected by a combination of soil moisture or flooding variables (thought to affect primarily tick vectors) and forest cover or NDVI variables (thought to affect primarily mammalian hosts). This research demonstrates the potential for modeling the distributions of microscopic tick-borne pathogens using coarse regional datasets and emphasizes the importance of forest cover and flooding as environmental constraints, as well as the importance of considering ecological variables at multiple spatial extents.

Comparison of serological methods and blood culture for detection of Trypanosoma cruzi infection in raccoons (Procyon lotor).

The indirect immunofluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were compared with blood culture for the detection of Trypanosoma cruzi infection in 83 raccoons (Procyon lotor) trapped in 4 counties of southeast Georgia. Both IFAT and ELISA detected 24 of 25 culture-positive samples (96% sensitivity). Cultures from 25 raccoons (30%) were positive for epimastigotes, whereas a total of 50 raccoons (60%) was seropositive by either the IFAT or ELISA. Forty-five of 83 serum samples (54%) were positive for anti-T. cruzi antibodies with the ELISA, and 47 were IFAT positive (57%). Forty-two of the 50 seropositive raccoons (84%) were seropositive by both tests. Endpoint titers of IFAT-positive samples were determined by testing doubling dilutions from 1:40 to 1:1280. High titers of 640 and 320 were observed for 4 raccoons trapped in 1 county (St. Catherines Island, Liberty County) and titers of 160 for 1-2 raccoons from each of the 4 counties sampled. IFAT titers and ELISA optical density values were positively correlated. Both serological tests have a high sensitivity and should be excellent tools for studying the prevalence of T. cruzi in wildlife populations.

Prevalence of trypanosome infections in dogs from Chagas disease endemic regions in Panama, Central America.

The prevalence of canine trypanosomosis was investigated in two Chagas disease endemic rural communities located in the central region of Panama. Serologic tests for Trypanosoma cruzi infection revealed a prevalence of 11.1%. Hemocultures coupled with PCR analysis demonstrated a Trypanosoma rangeli infection rate of 5.1%. An overall trypanosome infection index of 16.2% (16/99) was detected in this canine population. One dog had a mixed infection of T. cruzi and T. rangeli. Six of the trypanosome-infected dogs belong to people who were diagnosed of Chagas disease. We conclude that dogs from this rural area of Panama are frequently infected with trypanosomes transmitted by the sylvatic vector, Rhodnius pallescens, and suggest that dogs are important in the peridomestic transmission cycle of trypanosomes as reservoirs and hosts. The epidemiological implications of these findings are discussed.

Genetic characterisation of Toxoplasma gondii in wildlife from North America revealed widespread and high prevalence of the fourth clonal type.

Little is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study wild animals, from the USA were examined for T. gondii infection. Tissues of naturally exposed animals were bioassayed in mice for isolation of viable parasites. Viable T. gondii was isolated from 31 animals including, to our knowledge for the first time, from a bald eagle (Haliaeetus leucocephalus), five gray wolves (Canis lupus), a woodrat (Neotoma micropus), and five Arctic foxes (Alopex lagopus). Additionally, 66 T. gondii isolates obtained previously, but not genetically characterised, were revived in mice. Toxoplasma gondii DNA isolated from these 97 samples (31+66) was characterised using 11 PCR-restriction fragment length polymorphism (RFLP) markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). A total of 95 isolates were successfully genotyped. In addition to clonal Types II, and III, 12 different genotypes were found. These genotype data were combined with 74 T. gondii isolates previously characterised from wildlife from North America and a composite data set of 169 isolates comprised 22 genotypes, including clonal Types II, III and 20 atypical genotypes. Phylogenetic network analysis showed limited diversity with dominance of a recently designated fourth clonal type (Type 12) in North America, followed by the Type II and III lineages. These three major lineages together accounted for 85% of strains in North America. The Type 12 lineage includes previously identified Type A and X strains from sea otters. This study revealed that the Type 12 lineage accounts for 46.7% (79/169) of isolates and is dominant in wildlife of North America. No clonal Type I strain was identified among these wildlife isolates. These results suggest that T. gondii strains in wildlife from North America have limited diversity, with the occurrence of only a few major clonal types.