Transmission of Salmonella between wildlife and meat-production animals in Denmark.

AIMS: To investigate the transmission of Salmonella spp. between production animals (pigs and cattle) and wildlife on production animal farms in Denmark. METHODS AND RESULTS: In the winter and summer of 2001 and 2002, 3622 samples were collected from Salmonella-infected and noninfected herds of pigs and cattle and surrounding wildlife. Salmonella was detected in wildlife on farms carrying Salmonella-positive production animals and only during the periods when Salmonella was detected in the production animals. The presence of Salmonella Typhimurium in wild birds significantly correlated to their migration pattern and food preference. CONCLUSIONS: Salmonella was transmitted from infected herds of production animals (cattle and pigs) to wildlife that lived amongst or in close proximity to them. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella in animal food products is associated with the occurrence of Salmonella in primary animal production. Strategies to control the introduction and spread of infection should include wildlife management, as the nearby wildlife may act as reservoirs for Salmonella spp. and/or may be passive carriers of the bacteria.

Monoclonal antibodies to Toxoplasma gondii strain 119 identify recently isolated Danish strains as one group.

Four mAb raised against the Danish Toxoplasma gondii strain 119, were selected by screening hybridoma supernatants by indirect immunofluorescence against tachyzoites of the RH strain in order to obtain strain-restricted markers. Strain restriction extended beyond discrimination of the 119 and RH strains, as demonstrated on a further six T. gondii reference strains [BK and GT1 (group A), NTE and 561 (group B), and NED and C56 (group C)]. The bradyzoite-specific mAb, 4.3, reacted to the GT1, NTE and 561 strains, but not to the BK, NED or C56 strains. The tachyzoite-specific mAb, 4.25, reacted to all strains tested except the RH strain, while mAb 5.1 reacted to tachyzoites of strains NTE and 561, but not to those of the BK, GT1, NED or C56 strains. Monoclonal antibody 5.15 reacted with the same strain restriction as monoclonal antibody 5.1, but to bradyzoites as well as tachyzoites. A T. gondii strain collection representative for a small geographic area (Denmark) was established within a short time span from a variety of animal species. Using the mAb as typing reagents to this Danish strain collection, all 36 animal and two human strains were identified as having the same reaction pattern as strains 119, NTE and 561.

Serosurvey for antibodies to Borrelia burgdorferi in Danish dogs.

To estimate the regional presence of B. burgdorferi we performed a serosurvey for antibodies to B. burgdorferi in 205 healthy dogs from representative areas of Denmark. Blood samples were collected from November 1986 to March 1987. Twenty dogs bred for research purposes served as negative controls. IgG-antibodies to the B. burgdorferi strain DK-ECM 1 were measured by indirect immunofluorescens assay. Antibody titers ranged from nonreactive to 640. In 33 dogs (16.1%) titres were greater than or equal to 80, indicating exposure to B. burgdorferi. There were no significant differences in the prevalence rate of seropositive dogs due to place of residence, sex or age. The results indicate that dogs are exposed to tickborne B. burgdorferi in most areas of Denmark.

Disseminated histoplasmosis in a badger (Meles meles) in Denmark.

We report the first case of disseminated histoplasmosis in an animal in Scandinavia. Yeast cells compatible with those of Histoplasma capsulatum var. capsulatum were found in the skin, liver, spleen, a kidney, and a lymph node of a wild badger (Meles meles). The diagnosis was confirmed by electron microscopy and immunofluorescence staining of the yeast cells in tissue sections.

Astrovirus epidemiologically linked to pre-weaning diarrhoea in mink.

Diarrhoea and excessive secretion from the cervical apocrine glands in young, suckling mink kits is a well-known, but poorly defined, syndrome often referred to as "sticky", "greasy", or "wet" kits. We have performed a case-control study, at farm level as well as at mink kit level, in Denmark and Sweden to investigate whether enteric virus infections may be a risk factor in the development of pre-weaning diarrhoea. Tissue samples from the enteric tract of 180 sacrificed mink kits were analysed histologically. Faecal contents were examined by electron microscopy (EM). Astrovirus was detected in abundance and found to be a significant risk factor both at farm level (OR=21.60, p<0.001) and at mink kit level (OR=7.95, p<0.001). Other factors, i.e. low body weight, coccoid bacteria adherent to the enteric villi, and presence of calicivirus were also shown to increase the risk of pre-weaning diarrhoea, although with less impact than astrovirus.

Serogroups and antimicrobial susceptibility among Escherichia coli isolated from farmed mink (Mustela vison Schreiber) in Denmark.

Escherichia coli is commonly found in outbreaks of diarrhoea in mink during the production season although its role as a primary causal organism remains unclear. The present study was undertaken to determine the serogroups and antimicrobial susceptibility of E. coli isolates from healthy and diarrhoeic mink. Rectal swabs were taken from healthy and diseased animals, on six different farms, once at the onset of disease and again approximately 2 weeks later. The swabs were subjected to bacteriological investigation; a total of 210 E. coli were isolated, 98 from healthy animals and 112 from diseased. All isolates were serotyped and MICs were determined for nine antimicrobial compounds. Non-haemolytic isolates numbered 147, whereas 63 were haemolytic. Both haemolytic and non-haemolytic isolates were isolated from both healthy and diseased animals.A wide range of serogroups was detected, the most frequent being O2 (11.0%), O78 (11.0%), O153 (7.1%), O25 (5.7%), O6 (4.8%), and O15 (4.8%), but diarrhoea was not associated with specific serogroups. All isolates were sensitive to enrofloxacin, neomycin, gentamicin and colistin. In contrast, considerable variations in susceptibility were found among the six mink farms, for tetracycline (0-46.4%, average 21.9), ampicillin (2.9-50.0%, average 23.3), spectinomycin (8.0-35.7%, average 21.9), sulfamethoxazole (8.6-57.7%, average 30.0) and trimethoprim (0-35.7%, average 9.5). Resistance to tetracycline was statistically more prevalent among haemolytic than among non-haemolytic strains.In conclusion, serogrouping and haemolysin testing failed to identify any association with diarrhoeal disease and antimicrobial resistance was highly variable between different mink farms.

Assessment of the aerobic faecal microflora in mink (Mustela vison Schreiber) with emphasis on Escherichia coli and Staphylococcus intermedius.

The present study was undertaken to investigate the culturable aerobic faecal microflora of mink from newborn until adulthood with emphasis on the potential pathogens Escherichia coli and beta-haemolytic coagulase positive staphylococci. Rectal swabs were taken from 10 healthy dams and their offspring on seven mink farms throughout the production season and a semi-quantitative enumeration of total E. coli and haemolytic E. coli, beta-haemolytic streptococci, beta-haemolytic coagulase positive staphylococci, total lactic acid bacteria, and enterococci was carried out in all samples using selective and non-selective media. Aerobic bacteria were cultured from close to 100% of the samples throughout the survey. Prevalence of E. coli isolates varied between 70 and 90% of the samples throughout the survey with a small decline at the end of the study period. The highest bacterial counts were found among recently weaned kits or kits in the early growth period (P<0.0012). Lactic acid bacteria and enterococci were isolated from more than 90% of all samples, while beta-haemolytic staphylococci were isolated from 20 to 70% of the samples. While beta-haemolytic staphylococci were dominant from birth and during the nursing period, counts of staphylococci gradually decreased during the nursing period and were outnumbered by E. coli during the growth season.

Studies on manifestations of canine distemper virus infection in an urban dog population.

An upsurge of canine distemper was recognized at the beginning of 1991 in the urban dog population of the Copenhagen area. The outbreak had the characteristics of a virulent morbillivirus introduction in a partly immune population, where the disease primarily was manifested in young individuals. Testing of single serum samples for the presence of canine distemper virus (CDV) IgM antibodies using an IgM ELISA confirmed current and recent CDV infections in an urban dog population, where the use of attenuated CDV vaccines was widespread. In 49 out of 66 sera from clinical cases suspected of canine distemper we detected CDV IgM antibodies, as compared to the detection of viral antigen by indirect immunofluorescence in 27 of 65 specimens of conjunctival cells. The antigenic make-up of isolates from acute and subacute clinical cases was investigated with a panel of 51 monoclonal antibodies directed against CDV and the related phocine distemper virus. The isolates exhibited an homogeneous reaction pattern and shared overall antigenic characteristics of the CDV prototype. The majority of cases were diagnosed among unvaccinated dogs and individuals with unknown or obscure vaccination record. However, severe clinical cases were also diagnosed in vaccinated individuals.

Fatal toxoplasmosis in five cats.

Fatal toxoplasmosis was diagnosed in five of 155 cats necropsied in Denmark during a 2 year period, thus representing 3.2% of the necropsied animals. Lesions were dominated by scattered necrotic foci in liver and spleen. Toxoplasma gondii was demonstrated primarily by an immunoperoxidase technique. The clinical signs were rhinitis, conjunctivitis, dyspnoea and neurological malfunction. The relatively high prevalence indicates that clinical and fatal toxoplasmosis, naturally acquired, seems more frequently occurring among cats than is generally assumed.

Toxoplasmosis in a colony of New World monkeys.

In a colony of New World monkeys five tamarins (Saguinus oedipus, Saguinus labiatus and Leontopithecus rosal. rosal.), three marmosets (Callithrix jacchus and Callithrix pygmaea) and one saki (Pithecia pithecia) died suddenly. The colony comprised 16 marmosets, 10 tamarins and three sakis. The main pathological findings were necrotic lesions in the lung, the intestine, and the liver. Histopathologically T. gondii parasites were observed in organs from the tamarins and the marmosets but not in the saki. Some considerations on epidemiology are presented. Preventive measures were directed against the bottom layer of the cages, on cockroach extermination, and on freezing of raw meat.

Seroprevalence of Toxoplasma gondii in Danish farmed mink (Mustela vison S.).

One hundred and ninety-five mink sera randomly selected from 17 Danish mink farms were evaluated for the presence of Toxoplasma gondii antibodies in the latex agglutination test. Six (3%) sera contained T. gondii antibodies in titres of 1:64 or more. The estimated 3% prevalence means that 300,000 mink out of a total mink population of ten million might be infected with Toxoplasma gondii. This large number of possible sero-positive mink in Denmark indicates that there exists a potential risk of acquiring toxoplasmosis by pelting mink.

Experimental infection with Toxoplasma gondii in farmed mink (Mustela vison S.).

Live Toxoplasma gondii tissue cysts (Strain 119) were administered orally to five mink (Mustela vison S.) and five mink were inoculated with a suspension of ultrasonicated Toxoplasma gondii trophozoites (RH-strain). Seroconversion was observed in all animals administered live T. gondii cysts indicating that infection was established. Likewise seroconversion was observed in three out of four animals administered ultrasonicated T. gondii trophozoites. Faecal shedding of oocysts of T. gondii was not demonstrated in any of the infected animals. Histologically chronic meningitis with calcification was seen in all animals. Cerebral T. gondii tissue cysts were detected in four animals administered live tissue cysts. The study demonstrates that mink can be experimentally infected with T. gondii, thus representing a potential infection source for man, when considering the pelting procedure.

Influence of farm, feed-producer and season on incidence of gastrointestinal disorders in Danish farm mink.

The distribution of gastrointestinal disorders in mink in Danish farms is presented based on data collected in a longitudinal design. The time at risk was from weaning in June until pelting in November. The occurrence of gastrointestinal disorders after weaning of the mink kits together with the distribution within farms was studied. The period of highest risk was in the months immediately after weaning (July and August). More than 17% of the farms recorded GI disorders among > or = 10% of pairs of animals. A multilevel statistical analysis showed that besides the effect of time after weaning, a significant part of the variation in the incidence could be attributed to the farm and feed-producer. The potential for bias is discussed.

Pasteurella multocida from outbreaks of avian cholera in wild and captive birds in Denmark.

An outbreak of avian cholera was observed among wild birds in a few localities in Denmark in 2001. The highest mortalities were among breeding eiders (Somateria mollissima) and gulls (Larus spp.). Pulsed-field gel electrophoresis (PFGE) was conducted using ApaI and SmaI as restriction enzymes and restriction enzyme analysis (REA) using HpaII. The Pasteurella multocida subsp. multocida strain isolated from birds in this outbreak was indistinguishable from a strain that caused outbreaks in 1996 and 2003. Most isolates from domestic poultry had other PFGE patterns but some were indistinguishable from the outbreak strain. Among 68 isolates from wild birds, only one PFGE and one REA pattern were demonstrated, whereas among 23 isolates from domestic poultry, 14 different SmaI, 12 different ApaI, and 10 different HpaII patterns were found. The results suggest that a P. multocida strain has survived during several years among wild birds in Denmark.

Verrucous endocarditis associated with Streptococcus bovis in mink (Mustela vison).

Between 1998 and 2001, mortalities due to verrucous endocarditis were experienced at several mink farms. Gram-positive cocci were isolated from the endocardium of all the animals examined but not always from other internal organs. Almost all the isolates were identified as Streptococcus bovis and only a few isolates belonged to other Streptococcus species. Typing by pulsed-field gel electrophoresis of a selection of isolates revealed several patterns and several different clones. Attempts to reproduce disease by the injection of cultures of a field isolate into healthy mink failed.

Distemper virus as a cause of central nervous disease and death in badgers (Meles meles) in Denmark.

During the summer of 2002 a distemper-like disease was observed in the free-ranging badger population in Denmark. It was characterised by grand seizures, abnormal behaviour and death; the badgers all had severe chronic pneumonia and some had non-suppurative encephalomyelitis. In this study, eight of the affected badgers were examined by gross pathological, histological, immunohistological, bacteriological, parasitological and virological methods, and were diagnosed with distemper; canine distemper virus was identified.

Mastitis in the lactating mink female (Mustela vison S.) and the development of "greasy kits".

"Greasy kits" is the result of a multifactorial disease complex with few known definitive aetiological factors. Mastitis has been hypothesized as a triggering factor although classical clinical signs of mastitis (rubor, tumor, dolor, calor) are rarely seen in lactating Danish mink females. In this study we sacrificed 2 groups of lactating mink females with a total of 78 mammary glands at day 19-30 after giving birth. The first group had raised normal mink kits while the other group had suffered severe attacks of greasy kits. We found no clinical or histopathological evidence of mastitis but isolated streptococci and staphylococci from 2 mammary glands in females raising greasy kits. These glands showed no clinical or histological signs of inflammation attributable to bacteria and we conclude that mastitis is not necessary for the generation of greasy kits.

Mortality in farmed mink: systematic collection versus arbitrary submissions for diagnostic investigation.

The distribution of diagnoses of mortality in mink submitted to the Danish Veterinary Laboratory (DVL) for diagnostic investigation in the calendar year 1997 was compared with the diagnoses of mortality in all dead mink collected at 4 selected farms (project farms) during the same period. A total of 1,015 submitted mink and 1,149 mink from the 4 project farms were subjected to post mortem investigation. The average size (breeding stock) of the project farms was larger than Danish farms on average. However, the distribution of colour types of the mink was comparable. The seasonal distribution of the material from project farms and that of the submissions were approximately the same. Differences in the distribution of diagnoses as well as recovered microorganisms were found, however, mainly related to the proportion of gastro-intestinal disorders and E. coli respectively. These proportions were negatively correlated. Overall the results showed that extrapolating diagnostic results of laboratory submissions to the population of farmed mink may be problematic, and more reliable methods for disease surveillance must be considered.

A study on the predilection sites of Trichinella spiralis muscle larvae in experimentally infected foxes (Alopex lagopus, Vulpes vulpes).

Studies were carried out on the predilection sites of Trichinella spiralis muscle larvae in experimentally infected arctic foxes (Alopex lagopus) and silver foxes (Vulpes vulpes) reared in cages. The highest number of larvae per gram tissue was found in the muscles of the legs, eyes, diaphragm, and tongue. The 2 fox species showed no significant differences with regard to predilection sites.

The fox (Vulpes vulpes) as a reservoir for canine angiostrongylosis in Denmark. Field survey and experimental infections.

Until recently, Angiostrongylus vasorum was not considered to be an endogenous Danish parasite, since demonstration of this worm had been confined to necropsy findings in 2 dogs, both of which had visited France. During the last 2 years, however, clinical cases have been diagnosed among a considerable number of Danish dogs, none of which had ever been outside Denmark. All these cases have occurred north of Copenhagen, where an endemic focus seems to exist. In this field survey A. vasorum was found for the first time in wild Danish red foxes. Furthermore, experimental infections showed that the parasite can be transferred between foxes and dogs. Consequently, the wild fox population must be considered a potential reservoir for transfer of A. vasorum to domestic dogs.