Roland Mattsson

Detection of mecC-Positive Staphylococcus aureus (CC130-MRSA-XI) in Diseased European Hedgehogs (Erinaceus europaeus) in Sweden

Recently, a novel mec gene conferring beta-lactam resistance in Staphylococcus aureus has been discovered. This gene, mecC, is situated on a SCCmec XI element that has to date been identified in clonal complexes 49, 130, 425, 599 and 1943. Some of the currently known isolates have been identified from animals. This, and observations of mecA alleles that do not confer beta-lactam resistance, indicate that mec genes might have a reservoir in Staphylococcus species from animals. Thus it is important also to screen wildlife isolates for mec genes. Here, we describe mecC-positive Staphylococcus aureus (ST130-MRSA-XI) and the lesions related to the infection in two diseased free-ranging European hedgehogs (Erinaceus europaeus). One was found dead in 2003 in central Sweden, and suffered from S. aureus septicaemia. The other one, found on the island of Gotland in the Baltic Sea in 2011, showed a severe dermatitis and was euthanised. ST130-MRSA-XI isolates were isolated from lesions from both hedgehogs and were essentially identical to previously described isolates from humans. Both isolates carried the complete SCCmec XI element. They lacked the lukF-PV/lukS-PV and lukM/lukF-P83 genes, but harboured a gene for an exfoliative toxin homologue previously described from Staphylococcus hyicus, Staphylococcus pseudintermedius and other S. aureus of the CC130 lineage. To the best of our knowledge, these are the first reported cases of CC130-MRSA-XI in hedgehogs. Given that one of the samples was taken as early as 2003, this was the earliest detection of this strain and of mecC in Sweden. This and several other recent observations suggest that CC130 might be a zoonotic lineage of S. aureus and that SCCmec XI/mecC may have originated from animal pathogens.

Water-blooming and toxin-producing cyanobacteria in Swedish fresh and bracish waters, 1981–1995

During the period 1981–1995 331 phytoplankton samples from 153stations in different parts of Sweden were analysed regardingspecies composition and possible toxicity. Toxins were indicated in156 samples which gives a percentage of 47. The toxicity wasconfirmed by mouse bioassay. All samples were collected fromwater-blooming lakes, mainly containing taxa belonging to thecyanobacterial genera Anabaena, Aphanizomenon, Microcystis and Planktothrix. In addition to informationabout occurrence of neuro- or hepatotoxins, knowledge about thedistribution of water-blooming lakes in the country was obtained aswell as about their predominating taxa. Microcystisaeruginosa, Anabaena flos-aquae and Aphanizomenonflos-aquae were the most frequently occurring taxa in the lakes.Among the taxa observed, nine different cyanobacterial species withcertainty produce toxins. Microcystis spp. most frequentlyproduced toxic blooms (only hepatotoxins). Anabaena spp.,however, caused the majority of waterblooms. The Anabaenaflos-aquae-group produced neurotoxins [incl. anatoxin-a(s)],neurotoxins + hepatotoxins or only hepatotoxins. Aphanizomenon flos-aquae was only recorded as toxin-producing inthree Swedish lakes, which may be compared with the conditions inFinland, where it was toxic in 36 bloom samples. Aspects are givenon toxicity versus lake acidification and trophic conditions, aswell as on health effects on animals and humanbeings.

Pathology of natural highly pathogenic avian influenza H5N1 infection in wild tufted ducks (Aythya fuligula)

Highly pathogenic avian influenza (HPAI) subtype H5N1 is an infectious systemic viral disease that results in high morbidity and mortality in poultry, and has been reported in a wide range of wild bird species during the last few years. An outbreak of HPAI H5N1 occurred in wild birds in Sweden in 2006 that affected several duck species, geese, swans, gulls, and raptors. Tufted ducks (Aythya fuligula) accounted for the largest number of positive cases and, therefore, were selected for more in-depth histologic and immunohistochemical evaluations. The main histologic lesions associated with the presence of avian influenza antigen were found in the brain, pancreas, and upper respiratory tract. Other tissues in which influenza antigen was variably found included liver, lung, adrenal glands, kidneys, and peripheral nerve ganglia. The current study describes the pathology and viral tissue targeting of H5N1 by using histology, polymerase chain reaction, and immunohistochemistry, and highlights the range and variation in the presentation of the natural disease in tufted ducks.

A previously unidentified Chorioptes species infesting outer ear canals of moose (Alces alces): characterization of the mite and the pathology of infestation.

BackgroundDuring the past decade, Chorioptes mites occupying the outer ear canals have been a common finding at routine necropsies of moose (Alces alces) in Sweden, but neither the taxonomy of the mites nor lesions from the infestation have been investigated. In this study, the mites are characterized by morphological and molecular techniques, and the histopathology of the skin of the outer ear canal is described.MethodsExternal auditory meatuses from 53 necropsied moose were examined for the presence of Chorioptes, and samples from outer ear canals were taken for histopathological and microbiological examination. A proportion of the mites from each moose was identified to species. The DNA was extracted from mites from three moose, and their ITS-2 sequences were determined; these sequences were compared phylogenetically to sequences from other Chorioptes taxa.ResultsChorioptes mites were found in 43 (81%) of the 53 moose. The mites had morphological and genetic characteristics distinct from those of C. texanus and C. bovis, the two species generally accepted within the genus. Morphology also did not argue for a diagnosis as C. crewei, C. mydaus or C. panda. On histopathology, lesions were characterized by a hyperplastic perivascular to interstitial dermatitis with epidermal hyperkeratosis and crust formation. Dermal inflammatory infiltrates were composed of mixed T- and B-lymphocytes, plasma cells and macrophages, whereas eosinophils were notably uncommon. Staphylococcus aureus was grown from the infested epidermis of five of 14 examined moose.ConclusionChorioptes mite infestation was frequently detected in the outer ear canals of moose in Sweden. The mites were evidently pathogenic, being associated with inflammatory lesions of the external auditory meatus. Our studies indicate infestations with a previously undescribed Chorioptes species.

Cyanobacterial extracts and microcystin-LR are inactive in the micronucleus assay in vivo and in vitro.

Cyanobacteria are sometimes widespread in lakes and can produce potent toxins, which can be dangerous for animals that drink the water, e.g. cattle and dogs. If the toxins are taken up by fish and other organisms in the food chain, or occur in drinking-water, they may pose a problem also for humans. Microcystin-LR, a hepatotoxic cyclic peptide, is one of the most frequently found cyanobacterial toxins. Data on the genotoxic potential of microcystin-LR and other cyanobacterial toxins are contradictory. Here we report results of the micronucleus assay carried out in vivo and in vitro with these toxins. To increase the sensitivity, we used the flow cytometry-based micronucleus assay in the mouse. In this study both pure microcystin-LR and cyanobacterial extracts originating from four different lakes in Sweden were analysed. Although doses up to near lethality were used and an average of 200,000 young erythrocytes, polychromatic erythrocytes, were analysed from each animal, no genotoxic effect was observed, nor could any effect be shown in the in vitro micronucleus study, using human lymphocytes. These results show that the low concentration of microcystins that now and then occur in drinking-water does not increase the cancer risk through chromosome breaks or mal-distribution of chromosomes.

Endoparasites in the endangered Fennoscandian population of arctic foxes ( Vulpes lagopus )

The Fennoscandian arctic fox (Vulpes lagopus) population is endangered due to overharvest and competition with the larger red fox (Vulpes vulpes). In this study, we have screened the population in Sweden for endoparasites by analysis of non-invasively faecal samples collected at reproductive dens during two summers, one with low food abundance (2008) and the other with high food abundance (2010). Eggs, larvae and oocysts of a total of 14 different endoparasites were identified with a species richness per inhabited den of 3.2 (CI95% ± 0.48) in 2008 and 2.7 (CI95% ± 0.72) in 2010. Capillariidae-like eggs was identified at 59% of the dens in 2008 and 57% in 2010 and Toxocara canis with 7% (2008) and 30% (2010); Toxascaris leonina with 93% (2008) and 65% (2010); Uncinaria stenocephala 65% (2008) and 39% (2010); Crenosoma vulpis 3% (2008) and 4% (2010); Trichuris sp. 7% (2008) and 4% (2010); Cystoisospora canis-like oocysts 28% (2008) and 26% (2010); Cystoisospora ohiensis-like oocysts 38% (2008) and 4% (2010); Eimeria sp. 7% (2008) and 9% (2010); Sarcocystis sp. 3% (2008) and 9% (2010); Taenia sp. 10% (2008) and 4% (2010); Mesocestoides sp. 3% (2008) and 0% (2010); Balantidium sp. 0% (2008) and 9% (2010) and Spiruroidea-like eggs 0% (2008) and 4% (2010). To our knowledge, Balantidium sp., Sarcocystis sp. and Trichuris sp. has never been described before in wild arctic foxes.

Tularaemia in Norwegian dogs.

We describe tularaemia in a Norwegian dog caused by Francisella tularensis subspecies holarctica. A Hamilton Hound and his owner developed tulaeremia after hunting an infected mountain hare (Lepus timidus). The dog showed signs of lethargy, anorexia and fever during a period two to four days after hunting and thereafter fully recovered. Its antibody titers increased 32-fold from one to three weeks post exposure. Thereafter, the titer declined and leveled off at moderate positive values up to one year after exposure (end of study). This is believed to be the first case report of clinical F. tularensis subspecies holarctica infection in a European dog. In 2011, enormous numbers of Norway lemmings (Lemmus lemmus) occurred in Finnmark, the northernmost county of Norway and many dogs caught and swallowed lemmings. Some of these dogs developed non-specific signs of disease and the owners consulted a veterinary surgeon, who suspected tularaemia. In order to investigate this hypothesis, serum samples from 33 dogs were examined for antibodies to F. tularensis. The dogs were allocated into three groups: Dogs from Finnmark that became sick (Group 1) or remained healthy following contact with lemmings (Group 2), and healthy control dogs from Oslo without known contact with lemmings (Group 3). All the serum samples were analyzed with a tube agglutination assay. Among dogs exposed to lemmings, 10/11 and 3/12 were antibody positive in Group 1 and Group 2, respectively, whereas none of the control dogs (n=10) were positive for antibodies against F. tularensis. These results strongly indicate that the non-specific disease seen in the dogs in Finnmark was linked to F. tularensis infection acquired through contact with lemmings.

Pearsonema (syn Capillaria) plica associated cystitis in a Fennoscandian arctic fox (Vulpes lagopus: a case report

The bladderworm Pearsonema (syn Capillaria) plica affects domestic dogs and wild carnivores worldwide. A high prevalence in red foxes (Vulpes vulpes) has been reported in many European countries. P. plica inhabits the lower urinary tract and is considered to be of low pathogenic significance in dogs mostly causing asymptomatic infections. However, a higher level of pathogenicity has been reported in foxes. A severe cystitis associated with numerous bladderworms was found in a captive arctic fox (Vulpes lagopus) originating from the endangered Fennoscandian arctic fox population. To our knowledge this is the first description of P. plica infection in an arctic fox.

ICP‐MS for Screening of Metal Ions in Algae and Waste Water

By using Inductively Coupled Plasma‐Mass Spectrometry (ICP‐MS) multi‐element analysis was performed on six toxic and one non‐toxic cyanobacteria and waste water. A method was developed using a single standard together with one SRM allowing multi‐element screening. This technique simplifies the calibration procedure and allows exploration of unforeseen elements in the mass spectrum in a couple of minutes. In this investigation, reports the concentration of Mn, Cu, Zn, Rb, Sr, Cd, Ba, La, Pb and U in the cyanobacteria. High concentration of Mn was observed in Microcystis viridis. In the mass spectrum of the waste water from the cleaning plant in Starogard, Poland, it was observed that the concentration of Pb in water was higher after seven days than 120 days.

Pathology of natural Francisella tularensis subsp. holarctica infection in two yellow-necked mice ( Apodemus flavicollis )

BackgroundTularemia is a zoonosis caused by the bacterium Francisella tularensis. It has a wide host range, which includes mammals, birds and invertebrates. F. tularensis has often been isolated from various species of small rodents, but the pathology in naturally infected wild rodent species has rarely been reported.Case presentationHerein, we describe the pathology of tularemia in two naturally infected wild yellow-necked mice (Apodemus flavicollis). To visualize F. tularensis subsp. holarctica, indirect immunofluorescence and immunohistochemistry were applied on tissue sections. Real time polymerase chain reaction detected the bacterium in samples from liver and spleen in both mice. The only finding at necropsy was splenomegaly in one of the mice. Histological examination revealed necrotic foci in the liver associated with mild inflammation in both mice. Immunohistochemistry and indirect immunofluorescence showed bacteria disseminated in many organs, in the cytoplasm of macrophages, and intravascularly.ConclusionsThe two yellow-necked mice died of an acute disease caused by tularemic infection disseminated to many organs. Further investigations of naturally infected small rodents are important to better understand the variability in pathological presentation caused by infection by F. tularensis subsp. holarctica, as well to elucidate the importance of small rodents as transmitters and/or reservoirs.