AIR SAC TREMATODES (CYCLOCOELIDAE STOSSICH, 1902) INFECTING BIRDS IN ZOOLOGICAL INSTITUTIONS IN THE UNITED STATES.

Air sac trematodes (Digenea: Cyclocoelidae) were detected in 23 avian species from eight aviaries in the United States. Most of the infected host species were passeriform birds, but a few species in other orders also were infected. Four species of adult flukes were encountered: Circumvitellatrema momota, Morishitium sp., Psophiatrema greineri, and Szidatitrema yamagutii. Findings from retrospective review of medical records, necropsy records, and author observations are presented. Potential terrestrial snail intermediate hosts were collected from three indoor aviaries. A high prevalence (47%) of larval trematode infections was demonstrated in one species of nonnative snail (Prosopeas achatinacea); one larva was isolated and matched to the adult species (C. momota) from birds using PCR. Problems with introducing potentially infected wild-caught birds into aviaries, and exchanging captive individuals between aviaries where they potentially may carry infections, are discussed.

Amphiorchis stacyi n. sp. (Digenea: Spirorchiidae) in the heart of a green turtle from Florida, USA and the literature review of Amphiorchis (Price, 1934).

The present paper reports the occurrence of the seventh species in the genus Amphiorchis (Digenea: Spirorchiidae) collected from the heart of a green turtle found in Florida, USA. A taxonomic key to the species of Amphiorchis and a literature review for the genus are presented.

Evidence of Diversity, Site, and Host Specificity of Sea Turtle Blood Flukes (Digenea: Schistosomatoidea: "Spirorchiidae"): A Molecular Prospecting Study.

Neospirorchis (Digenea: "Spirorchiidae") are blood flukes of sea turtles. Trematodes tentatively identified as Neospirorchis sp. infect various sites within sea turtles inhabiting waters of the southeastern United States, but efforts to obtain specimens adequate for morphologic study has proven difficult. Two genetic targets, the internal transcribed spacer region of the ribosomal RNA gene and the partial mitochondrial cytochrome c oxidase subunit I gene, were used to investigate potential diversity among parasite specimens collected from stranded sea turtles. Sequence data were obtained from 215 trematode and egg specimens collected from 92 individual free-ranging cheloniid sea turtles comprising 4 host species. Molecular analysis yielded more than 20 different genotypes. We were able to assign 1 genotype to 1 of the 2 recognized species, Neospirorchis pricei Manter and Larson, 1950 . In many examples, genotypes exhibited host and site specificity. Our findings indicate considerable diversity of parasites resembling Neospirorchis with evidence of a number of uncharacterized blood flukes that require additional study.

COCCIDIAL INFECTION OF THE ADRENAL GLANDS OF LEATHERBACK SEA TURTLES (DERMOCHELYS CORIACEA).

Histologic lesions incidental to the cause of death were observed in the adrenal glands of 17 subadult and adult leatherback sea turtles ( Dermochelys coriacea ) found dead or moribund on or near shore in North America. Round bodies, 250-300 µm in diameter composed of an outer capsule and large multinucleated cells surrounding a central mass of acellular material were distributed throughout the affected glands. Protozoal etiology was suspected based on some resemblance to coccidia; however, features diagnostic for coccidial infection were lacking in all but one case, which had a focal area of adrenalitis containing zoites. A novel eucoccidian partial 18S rRNA genetic sequence was consistently detected in adrenal glands with lesions. With the use of quantitative PCR, a specific area of the V4 region of the coccidian 18S gene was quantified in affected adrenal glands and correlated significantly with density of the histologic lesions. A second distinct, but closely related, 18S sequence was also amplified from the adrenal gland of one turtle and from a fecal sample containing unsporulated coccidian oocysts. The two 18S sequences identified from leatherback sea turtles form a clade within the family Eimeriidae. Further investigation is required to understand better the morphology of the life stages, life cycle, and potential effects of this coccidian parasite on adrenal function.

Ticks associated with domestic dogs and cats in Florida, USA.

Voluntary collections of ticks from domestic dogs and cats by veterinary practitioners across Florida, USA, were conducted over a 10 month period. Of the 1337 ticks submitted, five species of ixodid ticks were identified and included Rhipicephalus sanguineus, Amblyomma americanum, A. maculatum, Dermacentor variabilis, and Ixodes scapularis. Most ticks were collected from dogs (98.4%) with the most predominant species being R. sanguineus (94.3%). Of the ticks collected from cats (1.6%), A. americanum were the most common (74%). Only R. sanguineus were collected throughout the state, with the other species collected only in central and north Florida. The tick species collected from dogs and cats represent a risk to these domestic species as well as associated humans for a range of tick-borne diseases in Florida.

The influence of changing prey availability on the prevalence of Diphyllobothrium in river otters from Yellowstone National Park.

Parasite prevalence in predatory mammals is influenced by numerous factors including diet, sex, season, and habitat. We examined the effect of such factors on the prevalence of gastrointestinal helminths in North American river otters ( Lontra canadensis ) in Yellowstone Lake, Yellowstone National Park. Otters in this ecosystem have recently experienced a decline in their main prey, Yellowstone cutthroat trout ( Oncorhynchus clarkii bouvieri), and have, in turn, increased consumption of alternative foods. Helminths were found in 13.2% of otter fecal samples. The dominant parasite was a Diphyllobothrium sp. ( Diphyllobothrium ditremum or Diphyllobothrium dendriticum ), a cestode acquired from cutthroat trout. Truttaedacnitis truttae and Contracaecum spp. nematodes were incidental parasites in otter feces, and acanthocephalan eggs were found in 1 sample. The prevalence of trout remains and a Diphyllobothrium sp. in otter feces was higher during the cutthroat trout spawning season. A Diphyllobothrium sp. was more prevalent in the feces of female otters. There was no relationship between annual declines in the frequency of trout in otter feces and prevalence of parasites. Helminth prevalence and species richness in Yellowstone otters was low and likely reflects low predator densities and few intermediate hosts. This is the first study reporting the helminth fauna of river otters in the Greater Yellowstone Area and confirms the otter as a definitive host for Diphyllobothrium sp. in this region.

Fatal metacestode infection in Bornean orangutan caused by unknown Versteria species.

A captive juvenile Bornean orangutan (Pongo pygmaeus) died from an unknown disseminated parasitic infection. Deep sequencing of DNA from infected tissues, followed by gene-specific PCR and sequencing, revealed a divergent species within the newly proposed genus Versteria (Cestoda: Taeniidae). Versteria may represent a previously unrecognized risk to primate health.

Diagnostic stages of the parasites of the Florida manatee, Trichechus manatus latirostris.

Limited information is available on diagnostic stages of parasites in Florida manatees (Trichechus manatus latirostris). We examined 67 fecal samples from captive and wild manatees to define the diagnostic stages of the parasite fauna known to occur in Florida manatees. Parasite eggs were freshly extracted ex utero from identified mature helminths and subsequently characterized, illustrated, and matched to those isolated from fecal samples. In addition, coccidian oocysts in the fecal samples were identified. These diagnostic stages included eggs from 5 species of trematodes (Chiorchis fabaceus, Chiorchis groschafti, Pulmonicola cochleotrema, Moniligerum blairi, and Nudacotyle undicola), 1 nematode (Heterocheilus tunicatus), and oocysts of 2 coccidians (Eimeria manatus and Eimeria nodulosa).

Co-infection and cross-species transmission of divergent Hepatocystis lineages in a wild African primate community.

Hemoparasites of the apicomplexan family Plasmodiidae include the etiological agents of malaria, as well as a suite of non-human primate parasites from which the human malaria agents evolved. Despite the significance of these parasites for global health, little information is available about their ecology in multi-host communities. Primates were investigated in Kibale National Park, Uganda, where ecological relationships among host species are well characterized. Blood samples were examined for parasites of the genera Plasmodium and Hepatocystis using microscopy and PCR targeting the parasite mitochondrial cytochrome b gene, followed by Sanger sequencing. To assess co-infection, "deep sequencing" of a variable region within cytochrome b was performed. Out of nine black-and-white colobus (Colobus guereza), one blue guenon (Cercopithecus mitis), five grey-cheeked mangabeys (Lophocebus albigena), 23 olive baboons (Papio anubis), 52 red colobus (Procolobus rufomitratus) and 12 red-tailed guenons (Cercopithecus ascanius), 79 infections (77.5%) were found, all of which were Hepatocystis spp. Sanger sequencing revealed 25 different parasite haplotypes that sorted phylogenetically into six species-specific but morphologically similar lineages. "Deep sequencing" revealed mixed-lineage co-infections in baboons and red colobus (41.7% and 64.7% of individuals, respectively) but not in other host species. One lineage infecting red colobus also infected baboons, but always as the minor variant, suggesting directional cross-species transmission. Hepatocystis parasites in this primate community are a diverse assemblage of cryptic lineages, some of which co-infect hosts and at least one of which can cross primate species barriers.

Enteropathogens identified in cats entering a Florida animal shelter with normal feces or diarrhea.

OBJECTIVE: To determine the frequency of enteropathogens in cats entering an animal shelter with normal feces or diarrhea. DESIGN: Cross-sectional study. ANIMALS: 100 cats evaluated at an open-admission municipal animal shelter in Florida. PROCEDURES: Fecal samples collected within 24 hours after admission from 50 cats with normal feces and 50 cats with diarrhea were tested by fecal flotation, antigen testing, PCR assay, and electron microscopy for selected enteropathogens. RESULTS: 12 enteropathogens were identified. Cats with diarrhea were no more likely to be infected with ≥ 1 (84%) enteropathogens than were cats with normal feces (84%). Only feline coronavirus was significantly more prevalent in cats with diarrhea (58%) than in cats with normal feces (36%). Other enteropathogens identified in cats with and without diarrhea included Clostridium perfringens enterotoxin A (42% and 50%, respectively), Cryptosporidium spp (10% and 20%, respectively), Giardia spp (20% and 8%, respectively), Cystoisospora spp (14% and 10%, respectively), hookworms (10% and 18%, respectively), ascarids (6% and 16%, respectively), Salmonella spp (6% and 4%, respectively), astrovirus (8% and 2%, respectively), feline panleukopenia virus (4% and 4%, respectively), calicivirus (0% and 2%, respectively), and Spirometra spp (0% and 2%, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: In the present study, cats entered the shelter with a variety of enteropathogens, many of which are pathogenic or zoonotic. Most infections were not associated with diarrhea or any specific risk factors such as signalment, source, or body condition, making it difficult to predict which cats were most likely to be infected. It is not possible to test all shelter cats for all possible infections, so practical guidelines should be developed to treat routinely for the most common and important enteropathogens.

Enteropathogens identified in dogs entering a Florida animal shelter with normal feces or diarrhea.

OBJECTIVE: To determine the frequency of enteropathogens in dogs entering an animal shelter with normal feces or diarrhea. DESIGN: Cross-sectional study. ANIMALS: 100 dogs evaluated at an open-admission municipal animal shelter in Florida. PROCEDURES: Fecal samples were collected within 24 hours after admission from 50 dogs with normal feces and 50 dogs with diarrhea. Feces were tested by fecal flotation, antigen testing, PCR assay, and electron microscopy for selected enteropathogens. RESULTS: 13 enteropathogens were identified. Dogs with diarrhea were significantly more likely to be infected with ≥ 1 enteropathogens (96%) than were dogs with normal feces (78%). Only Clostridium perfringens enterotoxin A gene was significantly more common in dogs with diarrhea (64%) than in dogs with normal feces (40%). Other enteropathogens identified in dogs with and without diarrhea included hookworms (58% and 48%, respectively), Giardia spp (22% and 16%, respectively), canine enteric coronavirus (2% and 18%, respectively), whipworms (12% and 8%, respectively), Cryptosporidium spp (12% and 2%, respectively), ascarids (8% and 8%, respectively), Salmonella spp (2% and 6%, respectively), Cystoisospora spp (2% and 4%, respectively), canine distemper virus (8% and 0%, respectively), Dipylidium caninum (2% and 2%, respectively), canine parvovirus (2% and 2%, respectively), and rotavirus (2% and 0%, respectively). CONCLUSIONS AND CLINICAL RELEVANCE: Dogs entered the shelter with a variety of enteropathogens, many of which are pathogenic or zoonotic. Most infections were not associated with diarrhea or any specific dog characteristics, making it difficult to predict the risk of infection for individual animals. Guidelines for preventive measures and empirical treatments that are logistically and financially feasible for use in shelters should be developed for control of the most common and important enteropathogens.

Hematozoa and a new haemoproteid species from Cathartidae (New World vulture) in South Carolina.

A survey of turkey vultures (Cathartes aura) in South Carolina revealed the presence of a novel haemoproteid. Haemoproteus catharti n. sp. is described from the slides developed from the blood of this vulture. The new species is a thick, halteridial form with complete margins, and is considered distinct from the other recognized haemoproteids from the diurnal raptors in shape, pigment number, parasite outline, and host-family specificity. While reviewing blood films, a series of unusual immature schizonts of Plasmodium sp. were also observed, and these are illustrated, along with another distinct haemoproteid from old slides of poor quality produced from turkey vulture blood.

Mite-filled cyst on a Brown-headed Cowbird (Molothrus ater) in Florida, USA.

A large, partly pedunculated mass on the scapular area of a wild-caught captive Brown-headed Cowbird (Molothrus ater) consisted of a multiloculated keratin cyst inhabited by a new species of harpirhynchid mite (Harpirhynchus quasimodo). The mass did not interfere with flight or behavior. This is the first record of such an infestation of cowbirds in Florida.

Cutaneous periocular Habronema infection in a dromedary camel (Camelus dromedarius).

A 6-year-old castrated dromedary camel (Camelus dromedarius) presented with a non-healing, severely pruritic, ulcerative fibrotic plaque located at the medial canthus. Histological examination of surgical biopsies identified degenerating nematode larvae within eosinophilic granulomas. Treatment involved repeated debridement of the lesion, injectable ivermectin and anti-inflammatory therapies, and injectable and topical antibiotics. A specially constructed mask with goggles to prevent the camel from continuing to self-traumatize the eye and lesion was also placed. Full recovery occurred approximately 1 month after diagnosis. Because of the location of the lesion, time of year, the gross and microscopic characteristics of the lesion, the presence of a likely nematode larva and the response to treatment, a diagnosis of cutaneous habronemiasis was made.

Association of Wolbachia with heartworm disease in cats and dogs.

Although the presence of adult Dirofilaria immitis in the pulmonary arteries and its associated arteritis and thromboembolic disease can explain some of the manifestations of canine and feline heartworm disease, the cause of other findings remains unclear. Cats with D. immitis antibodies but lacking adult parasites in the pulmonary arteries frequently develop histological lesions of the airways, resulting in a condition termed Heartworm-Associated Respiratory Disease. All D. immitis parasites harbor Wolbachia pipientis bacteria and D. immitis-infected animals can have circulating Wolbachia antibodies and pro-inflammatory Wolbachia antigens (WSP) deposited in tissues. Little is known about the role that Wolbachia plays in lung disease of animals naturally infected with D. immitis. The purpose of this study was to determine the contribution of Wolbachia to the pathogenesis of natural heartworm disease in cats and dogs. We hypothesized that animals having sufficient Wolbachia burden to be detected in lung tissue by immunohistochemistry and/or polymerase chain reaction (PCR) would have more severe pulmonary disease than those with bacteria below the limits of detection. We further hypothesized that animals that were immunoreactive to pro-inflammatory WSP would have more severe pulmonary lesions than those that were seronegative for WSP antibodies. Blood and lung tissue samples were collected from cats and dogs representing three different D. immitis infection statuses: heartworm-free, heartworm-exposed, heartworm-infected. There was a positive but weak correlation between the magnitude of D. immitis antibody titers and WSP titers in cats (r=0.57, p<0.001) and in dogs (r=0.39, p<0.001). Pulmonary lesions were more common in HW-infected animals than in HW-free animals. Pulmonary arteriolar occlusion was more common in HW-infected cats (57%; p=0.003) than in HW-infected dogs (17%). Although pulmonary lesions were most common in HW-infected animals, there was no clear additive effect when either Wolbachia DNA/WSP was detected in lung tissue or when circulating Wolbachia antibodies were detected. There were no significant differences in the magnitude of pulmonary lesion scores within each HW-infection status group regardless of whether Wolbachia DNA/WSP or antibodies were detected. The relationship between Wolbachia and lung pathology in heartworm-infected animals remains to be determined. The lack of clear evidence for a role of Wolbachia in heartworm disease creates a dilemma for veterinarians treating animals in D. immitis-endemic areas. Although the indiscriminant use of antibiotics should be avoided, many clinicians prescribe doxycycline based on the favorable responses observed in human filarial diseases and promising results from the first published studies of doxycycline use in D. immitis-infected dogs.

Toxocara vitulorum in beef calves in North Central Florida.

In this study, we evaluated the prevalence of Toxocara vitulorum in beef calves in North Central Florida. Fecal samples from 433 calves under 9 months of age were analyzed for the presence of eggs in their feces. The prevalence in calves less than 3 months of age was 17.6%, 3-4 months of age was 0.4% and those 5-6 months old had a 0.9% prevalence. As expected, no eggs were detected in any calves older than 6 months. Calves were treated with fenbendazole (10% FBZ) at 5mg/kg after fecal samples were collected. Twenty calves that had T. vitulorum eggs in the feces were resampled 2 weeks after treatment to evaluate effectiveness of FBZ. No T. vitulorum eggs were seen in the feces of 17/20 (85%) of the calves that were sampled after FBZ treatment. FBZ was effective in 85% of calves treated for T. vitulorum infection in calves. We would like to make beef ranchers and veterinarians in the southern states aware that the prevalence of this parasite has greatly increased recently in northern Florida beef units.

Renal infection by a new coccidian genus in big brown bats (Eptesicus fuscus).

A novel coccidian parasite from the kidney of big brown bats (Eptesicus fuscus) is described. This coccidian (Nephroisospora eptesici nov. gen., n. sp.) was associated with a generally mild, focal or multifocal, well-demarcated cortical renal lesion less than 1 mm in diameter. The lesion represented cystic, dilated tubules with hypertrophied tubular epithelial cells and was present in the kidneys of 29 of 590 bats. Numerous coccidian parasites in various stages of development were present within the tubular epithelial cells and within the cyst lumina. Oocysts were collected from cystic dilated tubules. Thin-walled, sporulated ellipsoidal oocysts measuring an average of 18.9 x 20.8 microm were present in kidney tissue. The oocysts contained 2 sporocysts with 4 sporozoites. A polar body and a prominent oocyst residuum were present in the oocysts, but no micropyle, sporocyst residuum, or Stieda bodies were detected. Analysis of the 18S rRNA gene sequence put the parasite in the Sarcocystidae. The parasite is closely related to Besnoitia, Hammondia, Neospora, and Toxoplasma. Ultrastructural features, such as the presence of an apical complex in merozoites, support the identification of a coccidian. A new genus and species, Nephroisospora eptesicii, is proposed for this unusual coccidian in which the entire cycle is completed in the kidney of a single host; it has a membrane-like oocyst wall, sporogony occurs in the host rather than in the abiotic environment, and the positioning of the parasite by nucleic acid sequence indicates it to be closely allied to Sarcocystis and Besnoitia.

Blood parasites in birds from Madagascar.

Madagascar has long been recognized for its unique and diverse biota. In particular, significant effort has been made to establish baseline population data to better conserve the endemic avifauna. During field expeditions between 1993 and 2004, birds were mist-netted at 11 different sites, at elevations from 60 m to 2,050 m above sea level. Data on endemic status, forest type, and habitat preference were recorded. Thin blood films from 947 birds, belonging to 26 families and 64 species, were examined by light microscopy to determine the prevalence of blood parasites. Of these 947 birds, 30.7% were infected by at least one species of blood parasite, 26.8% of which were infected by more than one species. Species of Haemoproteus were the most prevalent (17.4%), followed by microfilariae (11.0%), Leucocytozoon spp. (9.4%), Plasmodium spp. (1.9%), Trypanosoma spp. (0.9%), and Babesia spp. (0.2%). Species level identifications confirmed the presence of 47 species of hemosporidians and trypanosomes, which is notably high and mirrors the diversity of their avian hosts. Eleven (23.4%) of these parasite species were new to science and thought to be endemic to the island. Significant differences in prevalence were observed by sample site, forest type (humid vs. dry), and habitat preference. Birds from all elevational zones sampled were infected, although not all parasite genera were present in each zone. Four of the six endemic avian families or subfamilies (Bernieridae, Brachypteraciidae, Philepittinae [Eurylaimidae], and Vangidae) were sampled and found to be parasitized. Of the families with the largest sample sizes, the Zosteropidae and Ploceidae had the highest prevalence of infection (65.6% and 49.3%, respectively). The vectors of hematozoan parasites in Madagascar are currently unknown. These results add to the current knowledge of avian parasitism in Madagascar and are of particular interest for the conservation of endemic species, as well as threatened or endangered populations.

Education of global veterinarians.

In 2003, the University of Florida (UF) College of Veterinary Medicine (CVM) created an Office of International Programs (OIP) in response to one of ten initiatives of the UF Strategic Plan: internationalization of the curriculum. The OIP has developed coursework that provides students with an opportunity for international exposure during the veterinary curriculum at three levels. In Level 1 (on campus) students can participate in a seminar series in global health: www.ufglobalhealth.org. This is an elective course offered to professional students at the UF Health Science Center (Dentistry, Medicine, Pharmacy, Public Health, and Veterinary Medicine). In Level 2 (abroad), students can participate in structured study abroad programs under the supervision of UF faculty and international scholars from collaborative institutions abroad. In Level 3 (on campus and abroad), students can participate in a certificate program in international veterinary medicine. This is a 15-credit program, parallel to the veterinary curriculum. By offering courses on campus and abroad, we want to empower the curriculum with a global perspective of the veterinary profession, as well as with a humanist education that can help students recognize the importance of respect for cultural differences and the reasons for different degrees of development and growth in the world. In addition, this paper presents the need for veterinary medicine and other disciplines in the health sciences to communicate with other disciplines in the social sciences and natural sciences to create development practitioners equipped with cross-disciplinary knowledge and skills needed to formulate, implement and evaluate solutions aimed at breaking the cycle of poverty and disease in low income societies. Finally, this paper makes a call to the American Veterinary Medical Association Council on Education to assess the need to recognize the importance of internationalization of the veterinary curriculum as a key standard for accreditation of colleges or schools of veterinary medicine.

Echinococcus oligarthrus cystic hydatidosis in Brazilian agouti (Dasyprocta leporina).

Echinococcus oligarthrus hydatid cyst disease was diagnosed in two imported wild-caught Brazilian agouti (Dasyprocta leporina). The cysts were found incidentally during routine examinations and never appeared to cause any apparent clinical signs. Identification of the cysts was confirmed via histopathology. Over a period of 2 yr, the cysts calcified but did not reabsorb. Treatment with praziquantal and albendazole did not appear to affect the cysts. Agoutis are intermediate hosts of E. oligarthrus in the wild, where jaguars (Panthera onca) and pumas (Puma concolor) act as definitive hosts. Caution is advised when working with E. oligarthrus-infected animals because of the zoonotic disease potential.