Mariann Chriél

A retrospective study on salmonella infection in Danish broiler flocks

Abstract A retrospective longitudinal study was conducted to identify risk factors associated with Salmonella enterica infection in Danish broiler production. The study was based on information in the antemortem database (AM database) where data were available for all broiler flocks slaughtered over the 2-year period from 1992 to 1993 in Denmark. The AM database contains information collected by the ante-mortem veterinarians, from the slaughterhouses, and from the salmonella examinations carried out at the National Veterinary Laboratory. The epidemiological unit was the individual broiler flock. The salmonella status of the flock was determined by examining the caecal tonsils from 16 3-week-old chickens from each flock. This procedure would detect a salmonella-infected flock, with a probability above 95%, if the prevalence is above 20%. Furthermore, the structure and quality of the collected data have been evaluated. Fourteen variables were selected for analysis by multivariable logistic regression. An increased risk of salmonella infection in the broiler flocks was associated with the biggest hatcheries and feedmill, with an increasing number of houses on the farm, if the preceding flock was infected, and if the flock was reared in the autumn. Additionally, the main variables of the model were analysed by including a random effect at the house level. This resulted only in minor changes of the parameter estimates.

Generalised linear mixed models analysis of risk factors for contamination of Danish broiler flocks with Salmonella typhimurium.

We present a retrospective observational study of risk factors associated with the occurrence of Salmonella typhimurium (ST) in Danish broiler flocks. The study is based on recordings from 1994 in the ante-mortem database maintained by the Danish Poultry Council. The epidemiological units are the broiler flocks (about 4000 flocks) which are clustered within producers. Broiler flocks with ST-infected parent stocks show increased risk of salmonella infection, and also the hatchery affects the salmonella status significantly. Among the rearing factors, only the use of medicine as well as the time of rearing, and the sampling method are significant. Epidemiological control would seem most efficient on starting at the top levels of the production hierarchy from which a major part of the ST contamination is derived. A secondary purpose of the study is to evaluate different statistical approaches and software for the analysis of a moderately-sized data set of veterinary origin. We compare the results from five analyses of the generalised linear mixed model (GLMM) type. The first observation is that the results agree reasonably well and lead to similar conclusions. A closer look reveals certain patterns of bias and estimation accuracy that correspond well with theoretical findings and practical experience reported in the statistical literature.

Endoparasites of the raccoon dog (Nyctereutes procyonoides) and the red fox (Vulpes vulpes) in Denmark 2009–2012 – A comparative study

Graphical abstract

Diversity and stability of Aleutian mink disease virus during bottleneck transitions resulting from eradication in domestic mink in Denmark

Aleutian mink disease (plasmacytosis) virus (AMDV) in domestic mink (Neovison vison) has been subject to eradication in Denmark since 1976. In 2001, approximately 5% of Danish mink farms were still infected and all were located in the northern part of the peninsula of Jutland. In the present study a total of 274 Danish isolates of AMDV collected during the two seasons of 2004 and 2005 were characterized by partial sequencing of the coding region of the non-structural (NS) proteins. Older AMDV isolates from Denmark, available, were also included. The Danish isolates represent a very homogenous cluster compared with Swedish, Finnish and Dutch isolates and seem to represent a minor fraction of the genetic diversity previously found in Denmark. Stability of nucleotide deviations reveals that the purifying selection of bottlenecks imposed on the AMDV population in Denmark by the stamping out policy for more than 6 years exceeds the rate of mutation driven diversity. Among the isolates from farms in northern Jutland two distinct types could be identified and within each of them a number of sub-types which were all useful in tracking spread of infections. Infection at a farm the preceding season was a predisposing risk parameter for disease outbreak at a farm, and strain identity substantiates the suggestion that inadequate disinfection is involved in the recurrence of outbreaks. In cases of new introductions to farms it is indicated that contact including transport between farms played a most significant role.

Risk factors for the hazard of lameness in Danish Standardbred trotters

A follow-up study focusing on health problems interfering with optimal training of Danish Standardbred trotters was conducted with the participation of seven professional trainers. Our aim was to estimate the incidence of health problems that cause interruptions of optimal training, and to identify associations between the hazard of lameness and selected risk factors. The study population was dynamic and contained data of 265 Standardbred trotters monitored during 5 months in 1997 and 1998. The horses were >or=2 years old. Optimal training was defined as when the horse followed scheduled training including fast-speed trotting. Interruption of optimal training could only be caused by health problems and castration.A total of 123 new events of interruption of optimal training caused by health problems were reported. Lameness (injury located to joints and tendons) was the most-frequent cause of interruption of optimal training: 84 events in 69 horses (0.09 events per horse-month). Respiratory diseases (16 events) and muscular problems (seven events) were the second and third most-frequent causes of interrupted training. The effects of trainer, gender, age-group, time with a trainer, participation in races and current month on the hazard of lameness were estimated in a multivariable Cox proportional-hazard model. The effects of trainer, gender and age-group were modelled as time-independent. The effects of time with a trainer, participation in races and the current month were modelled as time-dependent variables. Trainer affected the hazard of lameness. Geldings had higher hazard than mares, as did 3-year olds (compared to >4-year olds). Compared to the period where horses had been trained by the same trainer for >3 months, horses in the period 1.5-2.5 months after they had entered the training regime had higher risk of lameness (hazard ratio: 3.2; 95% CI: 1.1-9.9). Participation in races increased the hazard of lameness significantly in the 5 days after a races.

Effect of climate and farm environment on Campylobacter spp. colonisation in Norwegian broiler flocks.

Campylobacteriosis is the most frequently reported zoonosis in the EU. A recent report states that between 50% and 80% of the human campylobacteriosis cases could be attributed to broiler as a reservoir. The current study was conducted to investigate associations between the presence of Campylobacter spp. in Norwegian broiler flocks and factors related to the climate and the farm environment. Data from 18,488 broiler flocks from 623 different farms during 2002-2007 were included in the study. A logistic regression analysis was conducted where Campylobacter spp. status of a broiler flock at the time of slaughter was defined as the dependent variable and farm was modelled as a random effect. The following factors were found to increase the probability for a broiler flock to test positive for Campylobacter spp.: daily mean temperature above 6°C during the rearing period, private water supply, presence of other livestock farms within a distance of 2 km, presence of other broiler farms within a distance of 4 km with flocks positive for Campylobacter spp. within 30 days prior to slaughter, heavy rainfall 11-30 days prior to slaughter, region and year. Daily mean temperature below 0°C reduced the probability. The study emphasises the importance of the farm environment and the climate for the occurrence of Campylobacter spp. in broiler flocks. The farm environment is probably a part of the Campylobacter spp. pathway into and between broiler flocks where farmyard run-off and humans or flies entering the houses might constitute vehicles transporting the organism. Fly activity is temperature-driven and flies might be a part of the explanation of the increased risk for Campylobacter spp. related to increased temperature demonstrated in the study.

Implementation and validation of a sensitive PCR detection method in the eradication campaign against Aleutian mink disease virus

Aleutian mink disease virus (AMDV) is a severe progressive disease causing multiple different clinical syndromes in mink. In Denmark, the disease is notifiable and under official control. The control programme, based on serological screening, has confined successfully AMDV to the northern part of Denmark. However, re-infections and new introductions of virus into farms require a confirmatory virological test to verify the positive test results of single animals and ultimately to investigate disease transmission. A one step PCR amplifying a 374-base fragment of the NS1 gene of AMDV was compared to the counter-current immune electrophoresis (CIE) routinely used in the serological screening programme. Mink organs (n=299) obtained from 55 recently infected farms and 8 non-infected farms from 2008 to 2010 were tested by PCR, and the results were found to have a high correlation with the serological status of the mink. The relative diagnostic sensitivity of the PCR was 94.7%, and the relative diagnostic specificity was 97.9% when read in parallel with the CIE. PCR positive samples were sequenced and phylogenetic analysis revealed high similarity within the analysed AMDV strains and to AMDV strains described previously.

Wildlife Reservoirs of Canine Distemper Virus Resulted in a Major Outbreak in Danish Farmed Mink (Neovison vison)

A major outbreak of canine distemper virus (CDV) in Danish farmed mink (Neovison vison) started in the late summer period of 2012. At the same time, a high number of diseased and dead wildlife species such as foxes, raccoon dogs, and ferrets were observed. To track the origin of the outbreak virus full-length sequencing of the receptor binding surface protein hemagglutinin (H) was performed on 26 CDVs collected from mink and 10 CDVs collected from wildlife species. Subsequent phylogenetic analyses showed that the virus circulating in the mink farms and wildlife were highly identical with an identity at the nucleotide level of 99.45% to 100%. The sequences could be grouped by single nucleotide polymorphisms according to geographical distribution of mink farms and wildlife. The signaling lymphocytic activation molecule (SLAM) receptor binding region in most viruses from both mink and wildlife contained G at position 530 and Y at position 549; however, three mink viruses had an Y549H substitution. The outbreak viruses clustered phylogenetically in the European lineage and were highly identical to wildlife viruses from Germany and Hungary (99.29% – 99.62%). The study furthermore revealed that fleas (Ceratophyllus sciurorum) contained CDV and that vertical transmission of CDV occurred in a wild ferret. The study provides evidence that wildlife species, such as foxes, play an important role in the transmission of CDV to farmed mink and that the virus may be maintained in the wild animal reservoir between outbreaks.

Influenza A(H10N7) Virus in Dead Harbor Seals, Denmark

Since April 2014, an outbreak of influenza in harbor seals has been ongoing in northern Europe. In Denmark during June–August, 152 harbor seals on the island of Anholt were found dead from severe pneumonia. We detected influenza A(H10N7) virus in 2 of 4 seals examined.

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.