Kjell Handeland

Salmonellae in Avian Wildlife in Norway from 1969 to 2000

ABSTRACT Postmortem records of wild-living birds in Norway with laboratory-confirmed findings of salmonella infection were summarized for the period from 1969 to 2000. Salmonella spp. were isolated from 470 birds belonging to 26 species. The salmonella-positive birds included 441 small passerines, 15 gulls, 5 waterfowl, 4 birds of prey, 3 doves, and 2 crows. The bullfinch (Pyrrhula pyrrhula) was by far the most frequently recorded species (54% of the cases). Salmonella enterica serover Typhimurium was recovered from all cases except from one hooded crow (Corvus corone), which yielded serovar Paratyphi-B var. Java. Variant O:4,12 comprised 96% (451 cases) of all serovar Typhimurium isolates, including all the passerines, while variant O:4,5,12 accounted for the remaining 4% (18 cases). The occurrence of salmonellae in small passerines showed a distinct seasonality, with a peak in February and March. Plasmid profile analysis of 346 isolates of serovar Typhimurium O:4,12 detected six profiles, of which two comprised 66 and 28% of the isolates, respectively. Phage typing of 52 randomly selected isolates of serovar Typhimurium O:4,12 from passerines detected four types: DT 40 (54%), U277 (35%), DT 99 (6%), and DT 110 (4%).

Campylobacter spp., Salmonella spp., Verocytotoxic Escherichia coli, and Antibiotic Resistance in Indicator Organisms in Wild Cervids

AbstractFaecal samples were collected, as part of the National Health Surveillance Program for Cervids (HOP) in Norway, from wild red deer, roe deer, moose and reindeer during ordinary hunting seasons from 2001 to 2003. Samples from a total of 618 animals were examined for verocytotoxic E. coli (VTEC); 611 animals for Salmonella and 324 animals for Campylobacter. A total of 50 samples were cultivated from each cervid species in order to isolate the indicator bacterial species E. coli and Enterococcus faecalis/E. faecium for antibiotic resistance pattern studies. Salmonella and the potentially human pathogenic verocytotoxic E. coli were not isolated, while Campylobacter jejuni jejuni was found in one roe deer sample only. Antibiotic resistance was found in 13 (7.3%) of the 179 E. coli isolates tested, eight of these being resistant against one type of antibiotic only. The proportion of resistant E. coli isolates was higher in wild reindeer (24%) than in the other cervids (2.2%). E. faecalis or E. faecium were isolated from 19 of the samples, none of these being reindeer. All the strains isolated were resistant against one (84%) or more (16%) antibiotics. A total of 14 E. faecalis-strains were resistant to virginiamycin only. The results indicate that the cervid species studied do not constitute an important infectious reservoir for either the human pathogens or the antibiotic resistant microorganisms included in the study.SammendragUndersøkelse av fecesprøver fra hjortevilt for Campylobacter spp., Salmonella spp., verocytotoksike Escherichia coli og antibiotikaresistens hos indikatororganismer. Fecesprøver ble samlet inn i regi av Helseovervåkingsprogrammet for hjortevilt (HOP) fra kronhjort, rådyr, elg og villrein i løpet av jaktsesongene fra 2001 til 2003. Prøver fra i alt 618 dyr ble undersøkt for verocytotoksiske E. coli (VTEC), 611 dyr for Salmonella og 324 dyr for Campylobacter. For å studere antibiotikaresistens- mønstre ble indikatorbakteriene E. coli og Enterococcus faecalis / E. faeciumi forsøkt isolert fra til sammen 50 prøver fra hver dyreart. Salmonella og de potensielt humanpatogene verocytotoksiske E. coli ble ikke isolert, mens Campylobacter jejuni jejuni ble funnet i prøven fra ett eneste rådyr. Antibiotikaresistens ble påvist hos 13 (7,3%) av de 179 E. coli isolatene som ble testet. Av disse var åtte resistente mot bare en type antibiotika. Andelen resistente E. coli isolater var høgere hos villrein (24%) enn hos det øvrige hjorteviltet (2,2%). E. faecalis eller E. faeciumi ble isolert fra 19 prøver, men ingen av disse var fra villrein. Alle stammene var resistente mot ett (84%) eller flere (16%) antibiotika. Til sammen 14 E. faecalis-stammer var resistente mot bare virginiamycin. Resultatene indikerer at hjortevilt ikke utgjør et smittereservoar av betydning verken for de humanpatogene bakteriene som inngikk i studien, eller for antibiotikaresistente bakterier.

ANTIBODIES TO RUMINANT ALPHA-HERPESVIRUSES AND PESTIVIRUSES IN NORWEGIAN CERVIDS

A serologic survey revealed that Norwegian populations of free-ranging reindeer (Rangifer tarandus tarandus), roe deer (Capreolus capreolus), red deer (Cervus elaphus), and moose (Alces alces) have been exposed to alpha-herpesviruses and pestiviruses. A total of 3,796 serum samples collected during the period 1993–2000 were tested in a neutralization test for antibodies against bovine herpesvirus 1 (BHV-1) or cervid herpesvirus 2 (CerHV-2), and 3,897 samples were tested by a neutralization test and/or enzyme-linked immunosorbent assay for antibodies against bovine viral diarrhea virus (BVDV). Antibodies against alpha-herpesvirus were found in 28.5% of reindeer, 3.0% of roe deer, and 0.5% of red deer, while all moose samples were negative. In reindeer, the prevalence of seropositive animals increased with age and was higher in males than females. Antibodies against BVDV were detected in 12.3% of roe deer, 4.2% of reindeer, 2.0% of moose and 1.1% of red deer. The results indicate that both alpha-herpesvirus and pestivirus are endemic in reindeer and pestivirus is endemic in roe deer in Norway. The viruses may be specific cervid strains. Seropositive red deer and moose may have become exposed as a result of contact with other ruminant species.

EPIDEMIOLOGIC AND PATHOLOGIC ASPECTS OF SALMONELLA TYPHIMURIUM INFECTION IN PASSERINE BIRDS IN NORWAY

Septicemic salmonellosis caused by Salmonella Typhimurium 4, 12: i : 1, 2 was diagnosed in 94 (64.8%) of 145 small passerines comprising nine species, examined in Norway during 1999–2000. The birds were found dead at private feeding places throughout the country. The bullfinch (Pyrrhula pyrrhula), Eurasian siskin (Carduelis spinus), common redpoll (Carduelis flammea), and Eurasian greenfinch (Carduelis chloris) were the most frequently affected species. Pathologic findings in 94 carcasses included poor body condition (84%), enlarged spleen (73%), and necrosis of crop/esophagus (78%), liver (53%), spleen (46%), proventriculus (13%), and intestine (5.3%). Histologically, necrosis consisted of debris, fibrin, inflammatory cells, and aggregates of Gram-negative bacteria and occasionally giant cells. Based on information from questionnaires sick and dead birds were observed at feeding places from December to June, with a distinct peak during February and March. The duration of recorded outbreaks varied from less than 1 wk to 4 mo. In a separate study, 1,990 apparently healthy passerines caught at feeding places established for bird-ringing purposes were surveyed for cloacal carriage of Salmonella spp. Forty (2.0%) of the birds examined, representing sampling sites both in southern and northern parts of the country, harbored S. Typhimurium 4, 12: i : 1, 2 in their intestines. The carrier species largely reflected the species most often suffering from fatal infection.

Screening of Feral Pigeon (Colomba livia), Mallard (Anas platyrhynchos) and Graylag Goose (Anser anser) Populations for Campylobacter spp., Salmonella spp., Avian Influenza Virus and Avian Paramyxovirus

A total of 119 fresh faecal samples were collected from graylag geese migrating northwards in April. Also, cloacal swabs were taken from 100 carcasses of graylag geese shot during the hunting season in August. In addition, samples were taken from 200 feral pigeons and five mallards. The cultivation of bacteria detected Campylobacter jejuni jejuni in six of the pigeons, and in one of the mallards. Salmonella diarizona 14:k:z53 was detected in one graylag goose, while all pigeons and mallards were negative for salmonellae. No avian paramyxovirus was found in any of the samples tested. One mallard, from an Oslo river, was influenza A virus positive, confirmed by RT-PCR and by inoculation of embryonated eggs. The isolate termed A/Duck/Norway/1/03 was found to be of H3N8 type based on sequence analyses of the hemagglutinin and neuraminidase segments, and serological tests. This is the first time an avian influenza virus has been isolated in Norway. The study demonstrates that the wild bird species examined may constitute a reservoir for important bird pathogens and zoonotic agents in Norway.SammendragScreening av byduer (Colomba livia), stokkender (Anas platyrhynchos) og grågås (Anser anser) for Campylobacter spp., Salmonella spp., aviært influenza virus og aviært paramyxovirus.Til sammen 119 ferske avføringsprøver ble samlet fra grågås på trekk nordover i april, og kloakksvabere ble tatt fra 100 skrotter av samme fugleart som ble skutt under jakta i august. I tillegg ble det tatt prøver av 200 byduer og fem stokkender. Ved bakteriedyrking ble Campylobacter jejuni jejuni funnet hos seks av duene og hos en av stokkendene. Salmonella diarizona 14 : k : z53 ble funnet hos ei grågås, mens alle duene og stokkendene var negative for salmonella. Aviært paramyxovirus ble ikke påvist i noen av de undersøkte prøvene. Ei stokkand fra ei elv i Oslo ble funnet positiv for influensa A virus med RT-PCR og ved innokkulering i embryonerte egg. Isolatet som benevnes A/Duck/Norway/1/03, ble funnet å være av H3N8 type, basert på sekvensanalyser av hemagglutinin og neuraminidase genene, og ved serologiske undersøkelser. Dette er første gang aviært influensavirus har blitt isolert i Norge. Studien viser at de ville fugleartene som ble undersøkt kan utgjøre et reservoar for viktige fuglepatogene og zoonotiske agens i Norge.

MALIGNANT CATARRHAL FEVER IN FREE-RANGING CERVIDS ASSOCIATED WITH OVHV-2 AND CPHV-2 DNA

Pathologic lesions were summarized in 18 free-ranging cervids (15 moose [Alces alces], two roe deer [Capreolus capreolus], and one red deer [Cervus elaphus[) diagnosed with malignant catarrhal fever (MCF) after examination at the National Veterinary Institute, Oslo 1982–2005. Eye lesions (conjunctivitis, corneal opacity, fibrin clots in the anterior eye chamber) were the most frequent gross finding. Erosive-ulcerative mucosal lesions in the nose and mouth were also commonly found. Histopathology revealed a nonpurulent vasculitis and perivasculitis in the central nervous system (CNS) typical of MCF in 16 of the cases. The diagnosis in the remaining two animals was based upon histologic eye lesions consistent with MCF (CNS not available for examination). Polymerase chain reaction was run on samples from 15 individuals for evidence of MCF-virus DNA, and ovine herpesvirus-2 (OvHV-2) DNA was detected in five moose, one roe deer, and one red deer, and caprine herpesvirus-2 (CpHV-2) DNA was detected in two moose and one roe deer. Sera from 1,000 free-ranging cervids were tested for specific antibodies to MCF-associated viruses (MCFV) by competitive inhibition enzyme-linked immunosorbent assay. The seroprevalences were: red deer 5%, reindeer (Rangifer tarandus) 4%, roe deer 2%, and moose 0.4% (n = 250 for all four species). The results indicate that sheep and goat MCFV may cause serious disease in wild moose, roe deer, and red deer. The seropositive cervids most likely represent individuals infected with either OvHV-2 or CpHV-2, but may also reflect infections with other related MCFV.

Avian Influenza Virus Screening in Wild Waterfowl in Norway, 2005

Abstract The prevalence of influenza A virus infection, and the distribution of different subtypes of the virus, were studied in 604 geese and ducks shot during ordinary hunting 2005. The study was based upon molecular screening of cloacal swabs taken by the hunters. The sampling included the following species: greylag (Anser anser), mallard (Anas platyrhynchos), wigeon (Anas penelope), teal (Anas crecca), goosander (Mergus merganser), tufted duck (Aythya fuligula), common scoter (Melanitta nigra), goldeneye (Bucephala clangula), and red-breasted merganser (Mergus serrator). The samples found to be positive in the initial pan-influenza A virus reverse transcription-polymerase chain reaction (RT-PCR) were further subtyped by using a specific H5 RT-PCR and full-length RT-PCRs for the hemagglutinin (HA) and neuraminidase genes. None of the greylag samples (0/185) were positive for influenza A virus, whereas 19.1% of the ducks (80/419) were positive. The prevalences of influenza A virus in the different duck species were as follows: mallard, 20.4% (58/284); wigeon, 12.5% (8/64); teal, 30.9% (13/42); goosander, 0% (0/5); tufted duck, 0% (0/4); common scoter, 14.3% (1/7); goldeneye, 0% (0/11); and red-breasted merganser, 0% (0/2). H5N1 subtype was found in one mallard and H5N2 subtype in another mallard and one teal. Sequencing of the HA gene identified all three viruses as low-pathogenic strains, closely related to low-pathogenic H5 influenza A viruses evidenced in recent years in Sweden and the Netherlands. The other subtypes identified included H1N1, H2, H3N2, H3N8, H6N1, H6N2, H6N8, H8N4, H9N2, H11N9, and H12 in mallards; H3N2, H6N2, H6N8, and H9N2 in teals; and H6N2 in wigeons and common scoter.

LIVER CONCENTRATIONS OF COPPER, COBALT, AND SELENIUM IN WILD NORWEGIAN RED DEER (CERVUS ELAPHUS)

Liver samples from 245 wild red deer (Cervus elaphus) collected during the licensed hunting season in 2001 from five different locations in western Norway were analyzed for copper (Cu), cobalt (Co), and selenium (Se). The associations between these trace elements and geographical location, age group, and sex were studied. The median (and range of) liver concentrations (μg/g wet weight) for all the examined deer were: Cu 20 (1.7–103), Co 0.08 (<0.01–0.18), and Se 0.09 (0.04–1.0). The results indicate a generally low status of Cu and Se. In total, 15 (6%) red deer had deficient Cu levels (<4 μg/g). For all three elements, the liver concentrations showed a significant geographic variation. The geographic difference was most distinct for Cu. The lowest median Cu concentration was found in deer from the island Hitra, where 13% of the animals had deficient Cu levels. Significant differences between age groups were found for all elements, and generally, the adults (≥2.5 yr) had the highest levels. No significant sex differences within the various age groups were found, with three exceptions: female calves and adults had significantly higher Co levels than male deer, and adult males had significantly higher Se levels than adult females. The Cu and Se status of wild red deer in parts of Norway is low; however, the significance of this needs to be explored further.

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.

ANTIBODIES TO GRANULOCYTIC EHRLICHIA IN MOOSE, RED DEER, AND ROE DEER IN NORWAY

Serum samples from 104 moose (Alces alces), 124 red deer (Cervus elaphus) and 114 roe deer (Capreolus capreolus), collected from different counties in southern Norway from 1994 to 2000, were analysed by an indirect immunofluorescent antibody staining method for antibodies to Ehrlichia equi.The overall seroprevalences for granulocytic Ehrlichiaspp. in moose, red deer, and roe deer from Ixodes ricinusinfested counties were 43%, 55%, and 96%, respectively. Antibody prevalence was significantly higher in roe deer than in moose and red deer (P<0.001). Mean antibody titers (log10 ± SD) to E. equiin sera from moose, red deer, and roe deer were 1:1,497 (3.17 ± 0.646), 1:234 (2.37 ± 0.424) and 1:676 (2.83 ± 0.404), respectively. The present work indicates that all these wild ruminant species are exposed to granulocytic Ehrlichiain Norway.