Inger Sofie Hamnes

GIARDIA DUODENALIS CYSTS ISOLATED FROM WILD MOOSE AND REINDEER IN NORWAY : GENETIC CHARACTERIZATION BY PCR-RFLP AND SEQUENCE ANALYSIS AT TWO GENES

There are few genotyping studies of Giardia duodenalis isolates from cervid hosts, although a previous study suggested that cervids may be a source of infection for humans and cattle. Giardia duodenalis isolates collected from wild moose (Alces alces) and reindeer (Rangifer tarandus) in Norway during 2002 and 2003 were characterized by polymerase chain reaction–restriction fraction length polymorphism (PCR-RFLP) at the β-giardin gene, and sequence analysis at both the β-giardin and glutamate dehydrogenase (gdh) genes. All results suggested that these isolates (n=25) belonged to assemblage A. Three different restriction patterns were obtained with PCR-RFLP, one of which has previously been associated with assemblage A. At the β-giardin gene, sequences from six reindeer isolates and one moose isolate were identical to a previously published assemblage A sequence from G. duodenalis cysts isolated from dairy calves. The other 10 moose isolates could be divided into five groups, with between two and 14 single nucleotide polymorphisms (SNPs) from the published genotype A2. At the gdh gene, three different sequences were obtained, differing from each other by between one and 15 SNPs and which have all been previously published as genotype A1, but with different specific hosts. Grouping of the isolates based on the sequences from both genes gave complex results; whereas all the G. duodenalis isolates from reindeer grouped together, two moose isolates, which had identical sequences at the β-giardin gene, had sequences that differed from each other by 15 SNPs at the gdh gene. The results of these studies, together with the large Norwegian populations of these cervids and the amount of fecal matter they produce, indicate that moose and reindeer may be significant reservoirs of G. duodenalis infection in Norway, which may be of importance to veterinary and public health.

A longitudinal study on the occurrence of Cryptosporidium and Giardia in dogs during their first year of life.

BackgroundThe primary aim of this study was to obtain more knowledge about the occurrence of Cryptosporidium and Giardia in young dogs in Norway.The occurrence of these parasites was investigated in a longitudinal study by repeated faecal sampling of dogs between 1 and 12 months of age (litter samples and individual samples). The dogs were privately owned and from four large breeds. Individual faecal samples were collected from 290 dogs from 57 litters when the dogs were approximately 3, 4, 6, and 12 months old. In addition, pooled samples were collected from 43 of the litters, and from 42 of the mother bitches, when the puppies were approximately 1 and/or 2 months old.MethodsThe samples were purified by sucrose gradient flotation concentration and examined by immunofluorescent staining.Results128 (44.1%) of the young dogs had one or more Cryptosporidium positive samples, whilst 60 (20.7%) dogs had one or more Giardia positive samples. The prevalence of the parasites varied with age. For Cryptosporidium, the individual prevalence was between 5.1% and 22.5%, with the highest level in dogs < 6 months old, and declining with age. For Giardia, the individual prevalence was between 6.0% and 11.4%, with the highest level in dogs > 6 months old, but the differences between age groups were not statistically significant. Significant differences in prevalences were found in relation to geographic location of the dogs. Both parasites occurred at low prevalences in Northern Norway.ConclusionBoth Cryptosporidium and Giardia are common in Norwegian dogs, with Cryptosporidium more prevalent than Giardia. Prevalences of the parasites were found to be influenced by age, geographical location, and infection status before weaning.

Arctic parasitology: why should we care?

The significant impact on human and animal health from parasitic infections in tropical regions is well known, but parasites of medical and veterinary importance are also found in the Arctic. Subsistence hunting and inadequate food inspection can expose people of the Arctic to foodborne parasites. Parasitic infections can influence the health of wildlife populations and thereby food security. The low ecological diversity that characterizes the Arctic imparts vulnerability. In addition, parasitic invasions and altered transmission of endemic parasites are evident and anticipated to continue under current climate changes, manifesting as pathogen range expansion, host switching, and/or disease emergence or reduction. However, Arctic ecosystems can provide useful models for understanding climate-induced shifts in host-parasite ecology in other regions.

Evidence of Spread of the Emerging Infectious Disease, Finch Trichomonosis, by Migrating birds

Finch trichomonosis emerged in Great Britain in 2005 and led to epidemic mortality and a significant population decline of greenfinches, Carduelis chloris and chaffinches, Fringilla coelebs, in the central and western counties of England and Wales in the autumn of 2006. In this article, we show continued epidemic spread of the disease with a pronounced shift in geographical distribution towards eastern England in 2007. This was followed by international spread to southern Fennoscandia where cases were confirmed at multiple sites in the summer of 2008. Sequence data of the ITS1/5.8S/ITS2 ribosomal region and part of the small subunit (SSU) rRNA gene showed no variation between the British and Fennoscandian parasite strains of Trichomonas gallinae. Epidemiological and historical ring return data support bird migration as a plausible mechanism for the observed pattern of disease spread, and suggest the chaffinch as the most likely primary vector. This finding is novel since, although intuitive, confirmed disease spread by migratory birds is very rare and, when it has been recognised, this has generally been for diseases caused by viral pathogens. We believe this to be the first documented case of the spread of a protozoal emerging infectious disease by migrating birds.

Illegal Wildlife Imports More than Just Animals—Baylisascaris procyonis in Raccoons (Procyon lotor) in Norway

Abstract In autumn 2011, 11 illegally imported animals were seized from a farm in southern Norway. These included four raccoon dogs (Nyctereutes procyonoides), four raccoons (Procyon lotor), and three South American coatis (Nasua nasua), all considered alien species in Norway. An additional two raccoons had escaped from the farm prior to seizure. The seized animals were euthanized and postmortem examination revealed that the four raccoons had moderate to high numbers of the zoonotic nematode Baylisascaris procyonis in their intestines, ranging from 11 to 115 nematodes per small intestine, with a mean of 53. The identity of the nematodes was confirmed using molecular analysis of ITS-1, ITS-2, cytochrome C oxidase 1, and 18S. Echinococcus multilocularis was not detected in any of the 11 animals. Toxocara and Toxascaris sp. eggs were detected in the feces of two raccoons, and two coatis had coccidia oocysts (80 and 360 oocysts per gram). Domestic dogs and other wildlife on the farm had potential access to the animal pens. Given that the eggs can remain infective for years in the environment, local veterinary and health authorities will need to remain vigilant for symptoms relating to infection with B. procyonis.

Molecular identification of Sarcocystis halieti n. sp., Sarcocystis lari and Sarcocystis truncata in the intestine of a white-tailed sea eagle(Haliaeetus albicilla) in Norway.

An emaciated white-tailed sea eagle (Haliaeetus albicilla) from Western Norway was found and nursed briefly before it died. The necropsy revealed that the principal cause of death was an inflammation and occlusion of the bile ducts. A secondary finding was the presence in the intestinal mucosa of numerous sporulated Sarcocystis oocysts measuring 21.8–22.8 × 16.0–17.0 μm. The aim of this study was to identify these oocysts to species level using molecular methods. Genomic DNA was extracted from 10 mucosal scrapings containing oocysts and subjected to PCR amplification and sequencing of four DNA regions: the 18S and 28S rRNA genes, the ITS1 region and the cox1 gene. DNA of three previously known Sarcocystis spp. was identified, but only two of these, Sarcocystis halieti n. sp. and Sarcocystis lari, both employing sea birds as intermediate hosts, were considered to have used the sea eagle as a definitive host and to have formed oocysts in its intestine. The third species found, Sarcocystis truncata, employs red deer as intermediate hosts and seems to use felids as definitive hosts based on its phylogenetic position and prevalence. The sea eagle had probably recently ingested portions of one of the latter hosts (red deer or cat/lynx) containing stages (sarcocysts/oocysts) and thus DNA of S. truncata. The species S. halieti and S. lari could only be unambiguously separated from their most closely related congeners on the basis of their ITS1 sequences. This is the first report of Sarcocystis oocysts in sea eagles and the first identification to species level of Sarcocystis oocysts in any type of eagle. The sea eagle also acted as intermediate host of an unidentified Sarcocystis spp. as evidenced by the finding of six thin-walled sarcocysts in a histological section of cardiac muscle.

Field evaluation of anticoccidial efficacy: A novel approach demonstrates reduced efficacy of toltrazuril against ovine Eimeria spp. in Norway

Ovine Eimeria spp. infections cause reduced welfare, increased mortality, and substantial economic losses, and anticoccidials are crucial for their control. Recent reports of toltrazuril resistance in pigs, and anecdotal reports of reduced anticoccidial efficacy in lambs, necessitate evaluation of anticoccidial efficacy. Due to the substantial lifecycle differences between nematodes and coccidia, current WAAVP methods for assessing anthelmintic efficacy are not suitable for such evaluations. Faecal samples were collected from 8 pairs of twin lambs from 36 Norwegian sheep farms 6–8 days after turnout. One twin of each pair was then treated with 20 mg/kg toltrazuril and a second faecal sample from all lambs was collected 7–11 days later. Oocyst excretion rate in all samples was determined using McMasters. Suitability of treatment timing was investigated by evaluating the increase in mean log oocyst excretion in untreated lambs. Based on comparisons between groups, a threshold of ≥0.75 (13 farms) was used to identify farms where drug efficacy could be assessed with confidence, drug efficacy on farms with increases of ≥0.5 but <0.75 (7 farms) were evaluated with caution, and drug efficacy on farms with increases of <0.5 (16 farms) was not estimated. Reduction in oocyst excretion between samples from treated lambs compared with controls from the 20 farms with a threshold of ≥0.5 were then analysed using a generalised linear mixed model. The results were classified based on 95% CI obtained using parametric bootstrapping. Among these 20 farms, two exhibited reduced drug efficacy (upper 95% CI < 95%), 13 had good efficacy (lower 95% CI > 90%), and for 5 the results were inconclusive. This is the first evidence-based report of reduced anticoccidial efficacy in ovine Eimeria spp. Additionally, we highlight the problem of sub-optimal timing of treatment (16/36 farms), which could potentially result in incorrect conclusions being reached regarding lack of drug efficacy.