OPTIMIZATION OF FECAL FLOTATIONS IN MARINE PARASITOLOGY AND DETERMINATION OF THE SPECIFIC GRAVITY OF HELMINTH EGGS IN PINNIPEDS.

Recent studies have sought to optimize the fecal flotation procedure to improve the detection of helminth eggs in terrestrial domestic species. It is unclear, however, whether these efforts in optimization are applicable to parasite species of marine environments, and verification of veterinary diagnostic procedures is clinically important. It was hypothesized that the eggs belonging to the parasites of pinnipeds would have different specific gravities (SpG) than those belonging to terrestrial hosts. Fecal samples were collected from each of 25 stranded pinnipeds representing three species (Zalophus californianus, Phoca vitulina, Mirounga angustirostris), and modified double centrifugal flotations were performed on 1-g samples. Among the 22 California sea lions sampled, trematode, ascarid, and cestode eggs were detected in 17/22 (77%), 10/22 (45%), and 4/22 (18%) individuals, respectively. Sugar-gradient modified centrifugation flotations were then conducted on a subset of 10 samples from California sea lions to evaluate the distribution of eggs in fractions representing varying SpG. Higher numbers of ascarid eggs were found in fractions representing a lower SpG (1.00-1.15), whereas trematode eggs belonging to the genus Zalophotrema were found in significantly higher numbers in the fraction representing 1.25 (P < 0.05). In conclusion, the SpG of trematode, ascarid, and cestode eggs from pinnipeds appears to be similar to those from terrestrial hosts, but numerous factors may affect their ability to be detected using traditional diagnostic approaches. Further exploration into the nature of the variability noted may lead to improved diagnostics in marine parasitology.

Giardia and Cryptosporidium in resident wildlife species in Arctic Alaska.

Giardia and Cryptosporidium are zoonotic protozoan parasites that can infect humans and other taxa, including wildlife, often causing gastrointestinal illness. Both have been identified as One Health priorities in the Arctic, where climate change is expected to influence the distribution of many wildlife and zoonotic diseases, but little is known about their prevalence in local wildlife. To help fill information gaps, we collected fecal samples from four wildlife species that occur seasonally on the northern Alaska coastline or in nearshore marine waters-Arctic fox (Vulpes lagopus), polar bear (Ursus maritimus), Pacific walrus (Odobenus rosmarus divergens), and caribou (Rangifer tarandus)-and used immunofluorescence assays to screen for Giardia cysts and Cryptosporidium oocysts. We detected Giardia cysts in 18.3% and Cryptosporidium oocysts in 16.5% of Arctic foxes (n = 109), suggesting that foxes may be potentially important hosts in this region. We also detected Giardia cysts in a single polar bear (12.5%; n = 8), which to our knowledge represents the first such report for this species. Neither parasite was detected in walruses or caribou.

Seasonal distributions and other risk factors for Giardia duodenalis and Cryptosporidium spp. infections in dogs and cats in Chiang Mai, Thailand.

The objectives of this study were to explore risk factors associated with Giardia and Cryptosporidium infections in dogs and cats in Chiang Mai, Thailand, to describe the seasonal distributions of Giardia and Cryptosporidium prevalence, and to determine the potential for zoonotic transmission through genetic characterization of isolates. Fecal samples from 301 dogs and 66 cats were collected between August 2009 and February 2010. The presence of Giardia cysts and Cryptosporidium oocysts was determined using zinc sulfate centrifugal flotation and immunofluorescent assay (IFA). Genotype/species were determined by DNA sequence analyses of PCR products from Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and triosephosphateisomerase (tpi) and Cryptosporidium heat shock protein 70KDa (hsp70) and small subunit-rRNA (SSU-rRNA) genes. Information related to specific risk factors was collected from owners of each animal using a questionnaire. The risk factor data were analyzed for associations with Giardia and Cryptosporidium infections using logistic regression. The overall estimated prevalence of Giardia and Cryptosporidium in dogs was 25.2% and 7.6%, respectively and in cats, 27.3% and 12.1%, respectively. The estimated prevalence of Giardia infection in dogs in the rainy season (31.7%) was significantly higher than in the drier, winter season (17.2%) (p < 0.01). The estimated prevalence of Cryptosporidium infection in dogs and of Giardia and Cryptosporidium infections in cats was not associated with season (p > 0.05). Multivariable analysis indicated that Giardia cysts were more likely to be detected in fecal samples of dogs that resided in high-density environments, drank untreated water, were shedding Cryptosporidium oocysts, were having acute diarrhea or a history of chronic diarrhea, and were collected in the rainy season. All 19 Giardia PCR positive samples typed as G. duodenalis canine adapted genotypes (assemblages C or D). In cats, of six Giardia PCR positive samples, five typed as dog assemblages and one typed as assemblage AI. Of ten dogs with Cryptosporidium PCR positive samples, eight typed as C. canis, one as C. parvum (a zoonotic species) and one had both C. canis and C. parvum. Of three Cryptosporidium PCR positive samples in cats, one typed as C. felis and two typed as C. parvum. The presence of zoonotic G. duodenalis assemblage AI in a cat, and C. parvum in feces of dogs and cats suggests a potential role for a reservoir for zoonotic transmission. Whether or not these presences were from exposure to other animal or human hosts or environment are needed to be confirmed.

Prevalence of and risk factors associated with shedding of Cryptosporidium felis in domestic cats of Mississippi and Alabama.

The prevalence of and risk factors associated with the shedding of cryptosporidial oocysts were determined for domestic cats in northeastern Mississippi and northwestern Alabama. Cryptosporidial oocysts were found in a single fecal samples from 30 of 250 cats using a centrifugal concentration technique followed by an immunofluorescent assay. The odds of a cat shedding oocysts were increased when another cat shedding oocysts was present in the household. Logistic regression analysis showed only concurrent shedding of Giardia cysts to be significantly associated with the presence of Cryptosporidium oocysts in the feces. Oocysts from 12 cats were successfully genotyped, based on sequencing of a fragment of the 18S rRNA gene, all isolates were Cryptosporidium felis.

Disseminated Strongyloides stercoralis infection in a dog following long-term treatment with budesonide.

CASE DESCRIPTION: A 1.5-year-old 1.5-kg (3.3-lb) castrated male Pomeranian was examined because of a 10-month history of diarrhea characterized by hematochezia and weight loss and an acute onset of respiratory distress (ie, tachypnea and dyspnea). A presumptive diagnosis of inflammatory bowel disease had been made previously, and the dog had been treated with budesonide and tylosin but continued to have diarrhea and weight loss. CLINICAL FINDINGS: On initial examination, the dog was weak and slightly obtunded. Thoracic radiography revealed a moderate to severe, diffuse, unstructured interstitial pattern. Serum biochemical abnormalities consisted of mild hypoalbuminemia, hypoglycemia, hypocalcemia, hypomagnesemia, and hypocholesterolemia that were likely secondary to chronic gastrointestinal disease and malnutrition. Pyuria and moderate bacteriuria with a single live larva were found on microscopic evaluation of the urine sediment. Fecal examination revealed numerous nematode larvae; the morphology was consistent with first-stage, rhabditiform larvae of Strongyloides stercoralis. TREATMENT AND OUTCOME: A diagnosis of disseminated S stercoralis infection was made. The dog was treated with fenbendazole and ivermectin but developed respiratory collapse approximately 12 hours later and was euthanized because of the poor prognosis. Postmortem examination revealed S stercoralis in the lungs, small intestine, and kidney. CLINICAL RELEVANCE: Findings illustrated the importance of performing diagnostic testing, including routine fecal examination, to rule out infectious causes of diarrhea before beginning empirical treatment with glucocorticoids such as budesonide. Further, repeated fecal examinations, including Baermann tests, should be considered if a positive response to glucocorticoids is not observed.

Comparison of diagnostic techniques for detection of Giardia duodenalis in dogs and cats.

BACKGROUND: An evaluation of currently available in-clinic diagnostic tests for Giardia duodenalis infection of dogs and cats has not been performed. In addition, there is discordance among published diagnostic comparisons. The absence of a true gold standard for detecting Giardia duodenalis also complicates diagnostic evaluations. OBJECTIVES: To evaluate diagnostic tests commercially available in the United States for detecting Giardia duodenalis in dogs and cats, in comparison to a widely used reference test, the direct immunofluorescent assay (IFA), and also to compare the results of 2 methods of analysis: comparison of diagnostic tests to a reference test (IFA) and Bayesian analysis. ANIMALS: Fecal samples from a convenience sample of 388 cats and dogs located in Colorado, Oklahoma, and Virginia. METHODS: Fecal samples were tested for Giardia duodenalis by zinc sulfate centrifugal fecal flotation and 4 different commercial diagnostic immunoassays. Results were analyzed via Bayesian analysis and by comparison to the IFA as the reference test. RESULTS: Sensitivity and specificity by comparison to IFA was ≥82% and ≥90%, respectively, for all diagnostic tests in dogs and cats. When analyzed via Bayesian analysis, sensitivity and specificity were ≥83% and ≥95%, respectively. When ZnSO4 centrifugal fecal flotation results were combined with immunoassay results, there was no longer a significant difference between the sensitivities of the commercial in-clinic immunoassays. CONCLUSION AND CLINICAL RELEVANCE: The Bayesian analysis validates using IFA as the reference test. Differences in commercial in-clinic immunoassay sensitivities can be mitigated when the results are combined with ZnSO4 centrifugal fecal flotation results.

Ecotoxicoparasitology of the gastrointestinal tracts of pinnipeds: the effect of parasites on the potential bioavailability of total mercury (THg).

Acanthocephalans, cestodes, and some species of nematodes acquire nutrients from the lumen contents in the gastrointestinal (GI) tract of their definitive host. These parasites are exposed to toxicants, such as mercury (Hg), through passive or active feeding mechanisms; therefore, the focus of this study was to determine if there is an effect of parasites on the dietary availability of total mercury (THg) within piscivorous pinniped hosts. THg concentrations ([THg]) in selected host tissues, parasites, and GI lumen contents from 22 California sea lions (Zalophus californianus), 15 ringed seals (Phoca hispida), and 4 spotted seals (Phoca largha) were determined. Among all pinnipeds, [THg] in acanthocephalans of the large intestine were significantly higher than concentrations in other samples (host lumen contents, other parasites and host intestinal wall), irrespective of location within the host GI tract. δ15N values of parasites depended both on parasite group and location within the GI tract. δ15N values were consistently higher in parasites inhabiting the large intestine, compared to elsewhere in the GI tract, for both sea lions and seals. δ13C values in parasites did not differ significantly from host GI tissues. Based on both [THg] and stable isotope values, parasites are likely affecting the Hg bioavailability within the GI lumen contents and host tissues, and toxicant-parasite interactions appear to depend on both parasitic taxon as well as their location within the host intestine.

Prevalence and Multilocus Genotyping Analysis of Cryptosporidium and Giardia Isolates from Dogs in Chiang Mai, Thailand.

The occurrence and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis isolated from dogs in Chiang Mai, Thailand were determined. Fecal samples were collected from 109 dogs between July and August 2008. Cryptosporidium spp. infection was determined by immunofluorescent assay (IFA), PCR assays that amplify Cryptosporidium heat-shock protein 70 kDa (hsp70), and two PCR assays that amplify a small subunit-ribosomal RNA (SSU-rRNA). Giardiaduodenalis infection was identified using zinc sulfate centrifugal flotation, IFA, and four PCR assays that amplify the Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and generic and dog-specific assays of triosephosphate isomerase (tpi) genes. Overall prevalence of Cryptosporidium spp. and G.duodenalis was 31.2% and 45.9%, respectively. Sequence analysis of 22 Cryptosporidium-positive samples and 21 Giardia-positive samples revealed the presence of C.canis in 15, and C. parvum in 7, G. duodenalis Assemblage C in 8, D in 11, and mixed of C and D in 2 dogs. Dogs in Chiang Mai were commonly exposed to Cryptosporidium spp. and G. duodenalis. Cryptosporidium parvum can be isolated from the feces of dogs, and all G. duodenalis assemblages were dog-specific. Dogs could be a reservoir for a zoonotic Cryptosporidium infection in humans, but further studies will be required to determine the clinical and zoonotic importance.

Ecotoxicoparasitology: Understanding mercury concentrations in gut contents, intestinal helminths and host tissues of Alaskan gray wolves (Canis lupus).

Some gastrointestinal helminths acquire nutrients from the lumen contents in which they live; thus, they may be exposed to non-essential elements, such as mercury (Hg), during feeding. The objectives of this study were: 1) determine the total mercury concentrations ([THg]) in Gray wolves (Canis lupus) and their parasites, and 2) use stable isotopes to evaluate the trophic relationships within the host. [THg] and stable isotopes (C and N) were determined for helminths, host tissues, and lumen contents from 88 wolves. Sixty-three wolves contained grossly visible helminths (71.5%). The prevalence of taeniids and ascarids was 63.6% (56/88) and 20.5% (18/88), respectively. Nine of these 63 wolves contained both taeniids and ascarids (14.3%). All ascarids were determined to be Toxascaris leonina. Taenia species present included T. krabbei and T. hydatigena. Within the GI tract, [THg] in the lumen contents of the proximal small intestine were significantly lower than in the distal small intestine. There was a significant positive association between hepatic and taeniid [THg]. Bioaccumulation factors (BAF) ranged from <1 to 22.9 in taeniids, and 1.1 to 12.3 in T. leonina. Taeniid and ascarid BAF were significantly higher than 1, suggesting that both groups are capable of THg accumulation in their wolf host. δ13C in taeniids was significantly lower than in host liver and skeletal muscle. [THg] in helminths and host tissues, in conjunction with stable isotope (C and N) values, provides insight into food-web dynamics of the host GI tract, and aids in elucidating ecotoxicoparasitologic relationships. Variation of [THg] throughout the GI tract, and between parasitic groups, underscores the need to further evaluate the effect(s) of feeding niche, and the nutritional needs of parasites, as they relate to toxicant exposure and distribution within the host.

Prevalence of internal parasites in beef cows in the United States: Results of the National Animal Health Monitoring System's (NAHMS) beef study, 2007-2008.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System's (NAHMS) 2007-2008 beef study, 567 producers from 24 US States were offered the opportunity to collect fecal samples from weaned beef calves and have them evaluated for the presence of parasite eggs (Phase 1). Participating producers were provided with instructions and materials for sample collection. Up to 20 fresh fecal samples were collected from each of the 99 participating operations. Fresh fecal samples were submitted to one of 3 randomly assigned laboratories for evaluation. Upon arrival at the laboratories, all samples were processed for the enumeration of strongyle, Nematodirus, and Trichuris eggs using the modified Wisconsin technique. The presence or absence of coccidian oocysts and tapeworm eggs was also noted. In submissions where the strongyle eggs per gram exceeded 30, aliquots from 2 to 6 animals were pooled for DNA extraction. Extracted DNA was subjected to genus level polymerase chain reaction (PCR) identification for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. In this study, 85.6% of the samples had strongyle type, Nematodirus, and Trichuris eggs. Among the samples evaluated, 91% had Cooperia, 79% Ostertagia, 53% Haemonchus, 38% Oesophagostomum, 18% Nematodirus, 7% Trichuris, and 3% Trichostrongylus. The prevalence of coccidia and tapeworm eggs was 59.9% and 13.7%, respectively.

Pendant l'étude de 2007­2008 chez les bovins effectuée par le Système national de surveillance des maladies animales (NAHMS) du Département de l'agriculture des États-Unis (USDA), 567 producteurs provenant de 24 états américains se sont vus offrir l'opportunité de prélever des échantillons de fèces de veaux sevrés et de les faire analyser pour la présence d'oeufs de parasite (Phase 1). On a fourni aux producteurs participants les instructions et le matériel pour le prélèvement d'échantillon. Jusqu'à 20 échantillons de fèces fraiches furent prélevés de chacune des 99 opérations participantes. Les échantillons de fèces fraiches furent soumis de manière aléatoire pour évaluation à l'un des trois laboratoires participants. Suite à l'arrivée au laboratoire, tous les échantillons étaient traités pour énumération des strongles, de Nematodirus, et d'oeufs de Trichuris en utilisant la technique de Wisconsin modifiée. La présence ou l'absence d'ookystes de coccidie et d'oeufs de vers plats furent également notées. Dans les échantillons soumis et dont le nombre d'oeufs de strongles par gramme dépassait 30, des aliquots de 2 à 6 animaux étaient regroupés pour extraction de l'ADN. L'ADN extrait était soumis à une réaction d'amplification en chaine par la polymérase (PCR) pour une identification au genre de la présence d'Ostertagia, de Cooperia, d'Haemonchus, d'Oesophagostomum, et de Trichostrongylus. Dans la présente étude, 85,6 % des échantillons avaient des strongles, du Nematodirus, et des oeufs de Trichuris. Parmi les échantillons évalués, 91 % avaient du Cooperia, 79 % de l'Ostertagia, 53 % de l'Haemoncus, 38 % de l'Oesophagostomum, 18 % du Nematodirus, 7 % du Trichuris, et 3 % du Trichostrongylus. Les prévalences de coccidies et d'oeufs de vers plats étaient respectivement de 59,9 % et 13,7 %.(Traduit par Docteur Serge Messier).

Effectiveness of current anthelmintic treatment programs on reducing fecal egg counts in United States cow-calf operations.

During the United States Department of Agriculture (USDA) National Animal Health Monitoring System's (NAHMS) 2007-2008 beef study, producers from 24 states were offered the opportunity to evaluate their animals for internal parasites and for overall responses to treatment with anthelmintics. A lapse of 45 d was required between initial sampling and any previous treatments. Choice of anthelmintic (oral benzimidazoles, and both injectable and pour-on endectocides) was at the discretion of the producer so as not to alter the local control programs. Fresh fecal samples were collected from 20 animals, or from the entire group if less than 20, then randomly assigned to 1 of 3 participating laboratories for examination. Analyses consisted of double centrifugation flotation followed by enumeration of strongyle, Nematodirus, and Trichuris eggs (the presence of coccidian oocysts and tapeworm eggs was also noted). Where strongyle eggs per gram (epg) exceeded 30, aliquots from 2 to 6 animals were pooled for egg isolation and polymerase chain reaction (PCR) analysis for the presence of Ostertagia, Cooperia, Haemonchus, Oesophagostomum, and Trichostrongylus. Results from 72 producers (19 States) indicated that fecal egg count reductions were < 90% in 1/3 of the operations. All operations exhibiting less than a 90% reduction had used pour-on macrocyclic lactones as the anthelmintic treatment. While some of these less than expected reductions could have been the result of improper drug application, PCR analyses of the parasite populations surviving treatment, coupled with follow-up studies at a limited number of sites, indicated that less than expected reductions were most likely due to anthelmintic resistance in Cooperia spp. and possibly Haemonchus spp.

Pendant l'étude de 2007­2008 chez les bovins effectuée par le Système national de surveillance des maladies animales (NAHMS) du Département de l'agriculture des États-Unis (USDA), des producteurs provenant de 24 états américains se sont vus offrir l'opportunité de faire évaluer leurs animaux pour la présence de parasites internes et pour leur réponse globale à un traitement avec des anthelminthiques. Un délai de 45 j était requis entre l'échantillonnage initial et un traitement antérieur. Le choix d'un anthelminthique (benzimidazole oral, et des endectocides injectables et en solution à verser) était à la discrétion du producteur afin de ne pas altérer les programmes de contrôle locaux. Des échantillons de fèces fraiches ont été prélevés de 20 animaux, ou de tout le groupe si moins de 20, puis ils ont été acheminés de manière aléatoire à un des trois laboratoires participants pour fin d'examen. L'analyse consistait en une double centrifugation par flottaison suivie d'une énumération des strongles, de Nematodirus, et d'oeufs de Trichuris (la présence d'ookystes de coccidie et d'oeufs de vers plats fut également notée). Lorsque le nombre d'oeufs de strongles par gramme dépassait 30, des aliquots de 2 à 6 animaux étaient regroupés pour isolement des oeufs et et soumis à une réaction d'amplification en chaine par la polymérase (PCR) pour détecter la présence d'Ostertagia, de Cooperia, d'Haemonchus, d'Oesophagostomum, et de Trichostrongylus. Les résultats provenant de 72 producteurs (19 états) indiquent que les réductions dans le dénombrement des oeufs dans les fèces étaient de < 90 % dans le tiers des opérations. Toutes les opérations montrant une réduction de moins de 90 % avaient utilisé des lactones macrocycliques en solution à verser comme traitement anthelminthique. Alors que certaines de ces réductions moindres que prévues puissent être le résultat d'une mauvaise application du produit, les analyses par PCR des populations de parasites survivantes au traitement, combinées aux études de suivis à un nombre limité de sites, indiquent que les réductions moindres que prévues étaient fort probablement dues à la résistance aux anthelminthiques chez Cooperia spp. et possiblement Haemonchus spp.(Traduit par Docteur Serge Messier).

EVALUATION OF ANTHELMINTIC FISHMEAL POLYMER BAITS FOR THE CONTROL OF BAYLISASCARIS PROCYONIS IN FREE-RANGING RACCOONS (PROCYON LOTOR).

Baylisascaris procyonis is a common gastrointestinal parasite of raccoons (Procyon lotor) and is a zoonotic helminth with the potential to cause severe or fatal infection. Raccoons thrive in human-dominated landscapes, and the fecal-oral transmission pathway and lack of effective treatment make B. procyonis a serious threat to public health. The distribution of medicinal baits has emerged as a socially acceptable and cost-effective method for managing disease in free-ranging wildlife. We assessed the suitability of a mass-producible anthelmintic bait for B. procyonis mitigation by evaluating the willingness of free-ranging raccoons to consume anthelmintic baits and determining whether bait consumption successfully cleared B. procyonis infections from raccoons. Anthelmintic baits were modified from standard fishmeal polymer baits, the food attractant commonly used in oral rabies vaccine baits, with the introduction of 220 mg of pyrantel pamoate into the fishmeal mixture. We captured 16 naturally infected raccoons, presented one anthelmintic bait, and monitored B. procyonis infection over 90 d by screening feces for eggs. Treatment cleared B. procyonis infections for nine of 12 raccoons that consumed >10 g of the 15 g bait. We used remote cameras to monitor in situ patterns of bait consumption for anthelmintic baits relative to standard baits. Both anthelmintic and standard baits were rapidly consumed, with no differences in the rate of consumption between bait types. However, after bait contact, raccoons demonstrated a greater willingness to consume standard baits while ignoring anthelmintic baits more frequently (P = 0.06). Initial trials of anthelmintic baits show promise, although refinement in both dose and palatability is needed. At mass production scales, the addition of pyrantel pamoate to fishmeal polymer baits would be inexpensive, potentially making anthelmintic baits a viable management option when coupled with an oral rabies vaccine or used independently for B. procyonis mitigation.

Prevalence of select vector-borne disease agents in owned dogs of Ghana.

Ticks, sera and ethylenediaminetetraacetic acid (EDTA) blood were collected from dogs evaluated at the Amakom Veterinary Clinic in Kumasi, Ghana. Sera were evaluated for Dirofilaria immitis antigen and antibodies against Borrelia burgdorferi, Anaplasma phagocytophilum and Ehrlichia canis. Conventional polymerase chain reaction assays designed to amplify the deoxyribonucleic acid (DNA) of Ehrlichia spp. or Anaplasma spp. or Neorickettsia spp. or Wolbachia spp., Babesia spp., Rickettsia spp., Hepatozoon spp., Bartonella spp. and the haemoplasmas were performed on DNA extracted from EDTA blood and all positive amplicons were sequenced. This small survey shows that the following vector-borne pathogens are present in urban Ghanian dogs: Ehrlichia canis, Hepatozoon canis,Dirofilaria immitis and Anaplasma platys. Bartonella henselae was isolated from ticks but not from the dogs.

Mercury in gray wolves (Canis lupus) in Alaska: increased exposure through consumption of marine prey.

Mercury (Hg) bioaccumulates in the tissues of organisms and biomagnifies within food-webs. Gray wolves (Canis lupus) in Alaska primarily acquire Hg through diet; therefore, comparing the extent of Hg exposure in wolves, in conjunction with stable isotopes, from interior and coastal regions of Alaska offers important insight into their feeding ecology. Liver, kidney, and skeletal muscle samples from 162 gray wolves were analyzed for total mercury (THg) concentrations and stable isotopic signatures (δ(13)C, δ(15)N, and δ(34)S). Median hepatic THg concentrations were significantly higher in wolves with coastal access compared to wolves from interior Alaska. Stable isotope ratios, in conjunction with THg concentrations, provide strong evidence that coastal wolves are utilizing marine prey representing several trophic levels. The utilization of cross-ecosystem food resources by coastal wolves is clearly contributing to increased THg exposure, and may ultimately have negative health implications for these animals.

Filaria taxideae in striped skunks (Mephitis mephitis) of Colorado, USA, and commonly associated filarial dermatitis.

During 2007-09, we necropsied striped skunks (Mephitis mephitis) from Colorado, USA. Eight of 51 (16%) had severe infections with the subcutaneous filarid nematode Filaria taxideae, and four of the infected skunks (50%) had dermatitis that was histologically associated with parasite ova in the skin.

Use of macrocyclic lactones in cattle in the USA.

The use of macrocyclic lactones has become the main stay for the treatment of endo- and ectoparasites in the cattle industry. Here we review those drugs that are currently approved for use in cattle in the United States. The general efficacy, tissue distribution and toxicity of each drug formulation are discussed. Included is a discussion regarding the current status for nematode anthelmintic resistance in cattle populations within the United States. Also provided is a current summary of ecological effects of macrocyclic lactones residues in manure.

Baylisascaris procyonis in raccoons (Procyon lotor) from eastern Colorado, an area of undefined prevalence.

Baylisascaris procyonis is a zoonotic parasite that has been documented in raccoons throughout much of the United States; however, no published information on its occurrence is available for the transition zone from the Great Plains to the Rocky Mountains. Because this parasite can cause neural larva migrans and diffuse unilateral subacute neuroretinitis in humans (as well as other hosts), a more complete understanding of the distribution of this parasite seems warranted for public health reasons. The purpose of this study was to begin to fill in the gaps in our knowledge of the distribution of B. procyonis in an area of the US where there is, currently, no published information available. Fifty-three raccoons were collected throughout eastern Colorado during 2007-2010. Forty-six were examined by necropsy and seven by fecal flotation. Age (11 juveniles, 25 adults) and sex (16 males, 19 females) of the raccoons were recorded when intact carcasses were available. When available, feces were further processed for the detection of Giardia and Cryptosporidium using a direct fluorescent antibody detection method. B. procyonis was found in 31 of 53 raccoons (58.5%, 95% CI=44.1%, 71.9%). Mean intensity was 11.7 with a range of 1-49 worms per infected individual. There was no significant difference between age or sex, and the presence of ascarids or the number of ascarids. Cryptosporidium spp. oocysts and Giardia spp. cysts were detected in 11/44 (25%; 95% CI: 13.2, 40.3) and 3/44 (6.9%; 95% CI: 1.4, 18.7) raccoons, respectively. The genotype of the Giardia present could not be determined. The genotype of five of six cryptosporidial isolates was 100% homologous to the skunk genotype while the sixth was 100% homologous to Cryptosporidium parvum. Based on these results, both B. procyonis and Cryptosporidium spp. appear to be prevalent in raccoons of eastern Colorado.

Comparisons of mammalian Giardia duodenalis assemblages based on the ß-giardin, glutamate dehydrogenase and triose phosphate isomerase genes.

The objective of this study was to determine and compare the assemblages of Giardia duodenalis isolated from mammalian fecal samples using the ß-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) genes. A total of 202 samples, either submitted to the Veterinary Diagnostic Laboratory (Parasitology) at Colorado State University or part of ongoing research studies, were typed. A subset of 50 dog samples were also assessed by the tpi-D-specific primers. Of these, 183 were from dogs, 13 were from cats, two were from llamas, and one each was from a calf, an alpaca, a sheep, and a horse. The majority of the dogs (171 of 183 isolates) in this study were infected with only dog-adapted Assemblage C or D. The tpi-D-specific primers confirmed that 28 of the samples that typed as Assemblage D by the bg and gdh genes were also Assemblage D by the tpi-D-specific primers. Only 12 isolates were Assemblage A alone or Assemblage A and Assemblage C or D. Of the 13 cat isolates, seven were Assemblage F, two were Assemblage D, three were Assemblage A and 1 contained both Assemblages C and D. The calf isolate was Assemblage E (gdh, tpi) and the alpaca (bg, gdh), llamas (gdh), sheep (bg, gdh, tpi) and horse (tpi) isolates were all Assemblage A. When the assemblage could be determined for more than one gene, 91 of 117 dog isolates gave consistent results and 8 of 9 cat isolates gave consistent results.

Serologic surveillance of pathogens in a declining harbor seal (Phoca vitulina) population in Glacier Bay National Park, Alaska, USA and a reference site.

The harbor seal population in Glacier Bay National Park, Alaska, has declined by over 70% since 1992. The reasons for this decline are not known. We examined serum antibodies and feces for evidence of exposure to multiple pathogens in this population. We also studied harbor seals from a reference site on Kodiak Island. In 2007, we found antibodies against Leptospira spp. in 31% of specimens from harbor seals in Glacier Bay, but no detectable serum antibodies in samples from Kodiak. In 2008, no samples had detectable antibodies against Leptospira spp. No serum antibodies against Toxoplasma gondii, morbilliviruses, or presence of Cryptosporidium in fecal samples were detected. However, Giardia was found in 6% of the fecal samples from Glacier Bay. Our results indicate that the harbor seal population in Glacier Bay National Park could be immunologically naïve to distemper viruses and therefore vulnerable to these pathogens. Given the relatively low prevalence of antibodies and low titers, pathogens likely are not the reason for the harbor seal decline in Glacier Bay.