Chlamydia psittaci in garden birds in Sweden.

Increased numbers of human infections with Chlamydia psittaci have been associated with bird feeding activities in southern Sweden. Information on occurrence and genotype of C. psittaci in garden birds in Sweden is required to corroborate this finding but data are limited. Additionally, pathogenicity of C. psittaci for garden birds is poorly understood. In this study, C. psittaci infection was investigated in 275 garden birds representing 22 species submitted for wildlife disease surveillance between 2009 and 2019. PCR was used to detect C. psittaci DNA in liver and lung. Positive samples were genotyped, additional PCR was performed on feces, and tissues were examined microscopically. C. psittaci was found in six (2.2 %) birds; three great tits (Parus major), two feral (Columba livia) and one wood pigeon (Columba palumbus). Two great tits and the wood pigeon had inflammatory lesions associated with C. psittaci. In the great tits and wood pigeon, C. psittaci genotype A, the cause of most human cases, was detected. Genotype B, considered endemic in pigeons, was detected in the feral pigeons. Low incidence of C. psittaci in dead Swedish garden birds was similar to studies on apparently healthy Swedish birds. Pathological findings were consistent with C. psittaci being fatal in half of the positive birds, which also had higher bacterial loads in feces. This highlights the risk for human infection via infected garden birds, especially regarding great tits and pigeons.

Highly Pathogenic Avian Influenza (HPAI H5Nx, Clade 2.3.4.4.b) in Poultry and Wild Birds in Sweden: Synopsis of the 2020-2021 Season.

Highly pathogenic avian influenza (HPAI, Gs/Gd lineage) was introduced to Europe in 2005 and has since caused numerous outbreaks in birds. The 2020-2021 season was the hitherto most devastating when considering bird numbers and duration in Europe. Surveillance data, virologic results and epidemiologic investigations from the 2020-2021 outbreaks in Sweden were analysed. Subtypes H5N8 and H5N5 were detected on 24 farms with poultry or other captive birds. In wild birds, subtypes H5N8, H5N5, H5N1, H5N4, H5Nx were detected in 130 out of 811 sampled birds. There was a spatiotemporal association between cases in wild birds and poultry. Based on phylogeny and epidemiology, most of the introductions of HPAI to commercial poultry were likely a result of indirect contact with wild birds. A definite route of introduction to poultry could not be established although some biosecurity breaches were observed. No spread between farms was identified but airborne spread between flocks on the same farm was suspected. Our findings exemplify the challenges posed by the continuously changing influenza viruses that seem to adapt to a broader species spectrum. This points to the importance of wild bird surveillance, compliance to biosecurity, and identification of risk factors for introduction on poultry farms.

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

BACKGROUND: Tularemia 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 PRESENTATION: Herein, 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. CONCLUSIONS: The 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.

Gastrointestinal helminths of gray wolves (Canis lupus lupus) from Sweden.

As the Scandinavian wolf population is limited in size, it is only rarely subject to systematic studies on its disease biology, especially gastrointestinal parasites. Therefore, this study aims to describe the prevalence of gastrointestinal helminths of gray wolves hunted on a limited license as a part of a wildlife management program. Helminths of 20 wolves were examined post mortem by macroscopy and coprology. Intestinal worms of five species were recovered from 18 wolves (90%): Uncinaria stenocephala (90%), Taenia spp. (45%), Alaria alata (25%), and Mesocestoides spp. (5%). Of the taeniid specimens typed by multiplex PCR and sequencing of the cox1 gene, 25% belonged to Taenia hydatigena and 25% to Taenia krabbei. The overall species diversity was low compared to findings from wolves of the northern hemisphere. Fecal eggs of Eucoleus boehmi were detected in 12 wolves (60%). Fecal metastrongylid larvae were found in seven individuals (39%), but PCR analyses specific for Angiostrongylus vasorum were negative. The wolves were in good body condition suggesting that the parasite infestation had no negative impact on the general health of the examined wolves. Although some of the recovered parasite species have zoonotic or veterinary impact, it is not likely that the spare wolf population pose substantial threat to human or veterinary health.

GASTROINTESTINAL PARASITIC INFECTIONS IN DEAD OR DEBILITATED MOOSE ( ALCES ALCES) IN SWEDEN.

: Our objectives were to determine prevalence and intensity of gastrointestinal parasites of dead or euthanized emaciated moose in central and southern Sweden ( n=50) and to assess parasite intensity as a major contributing factor in the poor condition of these moose. All animals were infected and most had gastrointestinal nematodes. Seven parasite species were found in the abomasa and 10 species were found in the small intestine. Coinfections were commonly found in the abomasum ( Ostertagia antipini and Mazamastrongylus dagestanica) and in the small intestine ( Nematodirella alcidis and Trichostrongylus capricola). Moose had many nematodes; the highest numbers were 224,400 and 11,760, in the abomasum and in the small intestine, respectively. Fourteen moose had more than 40,000 gastrointestinal nematodes (excluding large intestine nematodes represented by Trichuris spp. and Oesophagostomum venulosum). Additionally, a moderate prevalence (36%) of protostrongylid larvae (dorsal spine larvae) and a low prevalence (2-4%) of protozoal infections were identified at microscopic examination of feces. Adult moose had significantly more parasites than did subadults. The results did not show parasite load as a single or a major cause of the moose mortality, but they provided an update on the species composition of helminth parasites in moose in Sweden and illustrated the extreme infection intensities that free-ranging moose can have.

Nodular onchocercosis in red deer (Cervus elaphus) in Sweden.

The presence of subcutaneous nodular onchocercosis was investigated at slaughter of 151 red deer (Cervus elaphus) (107 juveniles and 44 adults) between October-December 2015. The prevalence of subcutaneous nodules was 56%. Nodules were located in the lumbar region of the back in 96% of the cases, and 38% of the infected red deer had additional parasitic nodules in other body locations, such as rump, thorax, forelimbs and neck. The number of nodules per deer was 1-10 in two-thirds of the affected animals, and only 2% had more than 50 nodules. Significant differences in prevalence of nodule presence were found between adult (81%) and juvenile red deer less than two years of age (45%) (p = 0.0001). Species confirmation and identification was done on 14 randomly selected nodules taken from the lumbar region of 14 deer of different geographical origin, by sequencing the mitochondrial 12S, 16S and nad5 gene fragments. The sequences were more or less identical (12S = 99-100%; 16S = 93-100%; nad5 = 92-100%) to previously published sequences for Onchocerca flexuosa. The high prevalence of nodular onchocercosis caused by O. flexuosa in red deer in Sweden shows that the investigated areas in the southern third of Sweden have favorable vector (simuliids and ceratopogonids) conditions and suitable environment for the maintenance of the parasite. To our knowledge, this is the first systematic study of O. flexuosa in red deer in Sweden.

High occurrence of mecC-MRSA in wild hedgehogs (Erinaceus europaeus) in Sweden.

We investigated the occurrence of mecC-MRSA in wild hedgehogs (Erinaceus europaeus) in Sweden and characterized the obtained isolates. Samples from 55 hedgehogs from five counties of Sweden were cultivated selectively for MRSA and putative isolates were confirmed by real-time PCR detecting mecA, mecC, nuc and PVL genes. mecC-MRSA was confirmed in 35 (64%) animals from three geographically separated counties. Confirmed isolates were spa-typed and tested for antimicrobial susceptibility by broth microdilution. Eight different spa-types were identified (t843, t978, t3391, t9111, t10751, t10893, t11015, t15312) of which t843 (49%) was the most common. The spa-types t843, t3391 and t978 were found in isolates from two counties. The study shows that mecC-MRSA is common in wild hedgehogs in two counties of Sweden but occurs in hedgehogs also in other parts of the country. Our findings suggest that hedgehogs could be a reservoir for mecC-MRSA. In addition, similar spa-types of isolates from hedgehogs and isolates previously described in domesticated animals and in humans indicates transfer between these populations.

Systemic AA amyloidosis in the red fox (Vulpes vulpes).

Amyloid A (AA) amyloidosis occurs spontaneously in many mammals and birds, but the prevalence varies considerably among different species, and even among subgroups of the same species. The Blue fox and the Gray fox seem to be resistant to the development of AA amyloidosis, while Island foxes have a high prevalence of the disease. Herein, we report on the identification of AA amyloidosis in the Red fox (Vulpes vulpes). Edman degradation and tandem MS analysis of proteolyzed amyloid protein revealed that the amyloid partly was composed of full-length SAA. Its amino acid sequence was determined and found to consist of 111 amino acid residues. Based on inter-species sequence comparisons we found four residue exchanges (Ser31, Lys63, Leu71, Lys72) between the Red and Blue fox SAAs. Lys63 seems unique to the Red fox SAA. We found no obvious explanation to how these exchanges might correlate with the reported differences in SAA amyloidogenicity. Furthermore, in contrast to fibrils from many other mammalian species, the isolated amyloid fibrils from Red fox did not seed AA amyloidosis in a mouse model.

Novel hepatitis E like virus found in Swedish moose.

A novel virus was detected in a sample collected from a Swedish moose (Alces alces). The virus was suggested as a member of the Hepeviridae family, although it was found to be highly divergent from the known four genotypes (gt1-4) of hepatitis E virus (HEV). Moose are regularly hunted for consumption in the whole of Scandinavia. Thus, the finding of this virus may be important from several aspects: (a) as a new diverged HEV in a new animal species, and (b) potential unexplored HEV transmission pathways for human infections. Considering these aspects, we have started the molecular characterization of this virus. A 5.1 kb amplicon was sequenced, and corresponded to the partial ORF1, followed by complete ORF2, ORF3 and poly(A) sequence. In comparison with existing HEVs, the moose HEV genome showed a general nucleotide sequence similarity of 37-63% and an extensively divergent putative ORF3 sequence. The junction region between the ORFs was also highly divergent; however, two putative secondary stem-loop structures were retained when compared to gt1-4, but with altered structural appearance. In the phylogenetic analysis, the moose HEV deviated and formed its own branch between the gt1-4 and other divergent animal HEVs. The characterization of this highly divergent genome provides important information regarding the diversity of HEV infecting various mammalian species. However, further studies are needed to investigate its prevalence in the moose populations and possibly in other host species, including the risk for human infection.

Characterization of encephalitis in wild birds naturally infected by highly pathogenic avian influenza H5N1.

During the outbreak of highly pathogenic avian influenza (HPAI) H5N1 in Sweden in 2006, disease and mortality were observed in a number of wild bird species. Encephalitis was one of the most consistent and severe findings in birds submitted for postmortem examination. However, the distribution and severity of the inflammation varied among individuals. This study characterized the encephalitis and the phenotype of the cellular infiltrate in brains of 40 birds of various species naturally infected with HPAI H5N1. Brain sections stained with hematoxylin and eosin and immunostained for influenza A viral antigen were evaluated in parallel to brain sections immunostained with antibodies against T lymphocytes (CD3+), B lymphocytes (CD79a+), macrophages (Lectin RCA-1+), and astrocytes expressing glial fibrillary acidic protein. The virus showed marked neurotropism, and the neuropathology included multifocal to diffuse areas of gliosis and inflammation in the gray matter, neuronal degeneration, neuronophagia, vacuolation of the neuropil, focal necrosis, perivascular cuffing, and meningitis. Broad ranges in severity, neuroanatomical distribution, and type of cellular infiltrate were observed among the different bird species. Since neurotropism is a key feature of HPAI H5N1 infection in birds and other species and because the clinical presentation can vary, the characterization of the inflammation in the brain is important in understanding the pathogenesis of the disease and also has important diagnostic implications for sample selection.

Combining information from surveys of several species to estimate the probability of freedom from Echinococcus multilocularis in Sweden, Finland and mainland Norway.

BACKGROUND: The fox tapeworm Echinococcus multilocularis has foxes and other canids as definitive host and rodents as intermediate hosts. However, most mammals can be accidental intermediate hosts and the larval stage may cause serious disease in humans. The parasite has never been detected in Sweden, Finland and mainland Norway. All three countries require currently an anthelminthic treatment for dogs and cats prior to entry in order to prevent introduction of the parasite. Documentation of freedom from E. multilocularis is necessary for justification of the present import requirements. METHODS: The probability that Sweden, Finland and mainland Norway were free from E. multilocularis and the sensitivity of the surveillance systems were estimated using scenario trees. Surveillance data from five animal species were included in the study: red fox (Vulpes vulpes), raccoon dog (Nyctereutes procyonoides), domestic pig, wild boar (Sus scrofa) and voles and lemmings (Arvicolinae). RESULTS: The cumulative probability of freedom from EM in December 2009 was high in all three countries, 0.98 (95% CI 0.96-0.99) in Finland and 0.99 (0.97-0.995) in Sweden and 0.98 (0.95-0.99) in Norway. CONCLUSIONS: Results from the model confirm that there is a high probability that in 2009 the countries were free from E. multilocularis. The sensitivity analyses showed that the choice of the design prevalences in different infected populations was influential. Therefore more knowledge on expected prevalences for E. multilocularis in infected populations of different species is desirable to reduce residual uncertainty of the results.

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.

Diseases and mortality in free-ranging brown bear (Ursus arctos), gray wolf (Canis lupus), and wolverine (Gulo gulo) in Sweden.

Ninety-eight brown bears (Ursus arctos), 20 gray wolves (Canis lupus), and 27 wolverines (Gulo gulo), all free-ranging, were submitted to the National Veterinary Institute, Uppsala, Sweden, during 1987-2001 for investigation of diseases and causes of mortality. The most common cause of natural death in brown bears was infanticide. Infanticide also was observed in wolverines but not in wolves. Traumatic injuries, originating from road or railway accidents, were the most common cause of death in wolves and occurred occasionally in brown bears. Most wolverines were submitted as forensic cases in which illegal hunting/poaching was suspected. Sarcoptic mange was observed in several wolves but not in brown bears or wolverines. Sarcoptic mange most likely was acquired from infected red foxes (Vulpes vulpes) that were killed by wolves. Other parasites and infectious diseases were only found sporadically.

Investigations into shaking mink syndrome: an encephalomyelitis of unknown cause in farmed mink (Mustela vison) kits in Scandinavia.

An apparently novel neurological disease clinically characterized by shaking, tremors, seizures, staggering gait, and ataxia was first observed in farmed mink kits in Denmark in 2000 and subsequently in Sweden, Denmark, and Finland in 2001, and again in Denmark in 2002. Lymphoplasmacytic encephalomyelitis was found in the affected kits. The lesions were most severe in the brainstem and cerebellum and consisted of neuronal degeneration and necrosis, neuronophagia, focal and diffuse gliosis, perivascular cuffs formed by lymphocytes, plasma cells and macrophages, and segmental loss of Purkinje cells. Testing was conducted to determine the cause of the disease, including general virological investigations (virus culture, negative-staining electron microscopy, immunoelectron microscopy, polymerase chain reaction for herpesviruses, adenoviruses, pestiviruses, and coronaviruses), tests for specific viral diseases (canine distemper, Borna disease, Louping ill, West Nile virus infection, tick-borne encephalitis, Aleutian disease), tests for protozoa (Toxoplasma gondii, Neospora caninum, Encephalitozoon cuniculi), bacteria (general culture, listeria, Clamydophila psittaci), and intracerebral inoculation of neonatal mice. The results of all these investigations were negative. One group of 3 mink kits inoculated intracerebrally with brain homogenate of affected mink developed clinical signs and histological lesions similar to those observed in naturally infected mink. Based on the histopathological features, it is postulated that the disease is caused by a yet unidentified virus.