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Serogroups and antimicrobial susceptibility among
Escherichia coli
isolated from farmed mink (
Mustela vison
Schreiber) in Denmark
L. Vulfson
a,*
, K. Pedersen
a
, M. ChrieÂl
b
, K. Frydendahl
c
,T. Holmen Andersen
a
, M. Madsen
a
, H.H. Dietz
a
a
Danish Veterinary Laboratory, Hangùvej 2, DK-8200 Aarhus N, Denmark
b
Danish Fur Breeders Laboratory, Langagervej 74, DK-2600 Glostrup, Denmark
c
Danish Veterinary Laboratory, BuÈ lowsvej 27, DK-1790 Copenhagen V, Denmark
Received 17 April 2000; received in revised form 7 August 2000; accepted 28 August 2000
Abstract
Escherichia coli
is commonly found in outbreaks of diarrhoea in mink during the productionseason although its role as a primary causal organism remains unclear. The present study wasundertaken to determine the serogroups and antimicrobial susceptibility of
E. coli
isolates fromhealthy and diarrhoeic mink. Rectal swabs were taken from healthy and diseased animals, on sixdifferent farms, once at the onset of disease and again approximately 2 weeks later. The swabs weresubjected to bacteriological investigation; a total of 210
E. coli
were isolated, 98 from healthyanimals and 112 from diseased. All isolates were serotyped and MICs were determined for nineantimicrobial 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 withspeci®c serogroups. All isolates were sensitive to enro¯oxacin, neomycin, gentamicin and colistin.In contrast, considerable variations in susceptibility were found among the six mink farms, fortetracycline (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 amonghaemolytic than among non-haemolytic strains.In conclusion, serogrouping and haemolysin testing failed to identify any association withdiarrhoeal disease and antimicrobial resistance was highly variable between different mink farms.
#
2001 Elsevier Science B.V. All rights reserved.
Keywords: Escherichia coli
; Mink; Antimicrobial sensitivityVeterinary Microbiology 79 (2001) 143±153
*
Corresponding author. Tel.:
45-8937-2473; fax:
45-8937-2470.
E-mail address
: liv@svs.dk (L. Vulfson).0378-1135/01/$ ± see front matter
#
2001 Elsevier Science B.V. All rights reserved.PII: S0378-1135(00)00343-6
1. Introduction
In spite of considerable knowledge, on the basis of substantial pathological andmicrobiological experience, of
Escherichia coli
as a pathogen in humans and severalproduction animals, there is dearth of knowledge, of
E. coli
as the cause of infections inmink (
Mustela vison
Schreiber). To our knowledge, no elaborate work has hitherto beendone on disease determinants of
E. coli
isolated from fur animals. Infections with
E. coli
are common in mink production and occur as farm infections in all age groups throughoutthe season. These infections induce a higher mortality rate and a reduced fur quality thatcan cause substantial economic losses for the fur farmer and fur industry.
E. coli
is oftenisolated in association with the following well-de®ned disease complexes: (1) systemicinfections caused by
E. coli
in kits from 3 to 4 weeks of age resulting in septicaemia andconsiderable mortality (LoÈliger, 1970), (2) diarrhoea outbreaks at weaning (Hunter,1996a), (3)
E. coli
-associated diarrhoea occurring regularly from weaning till pelting(LoÈliger, 1970), and (4) lung infections throughout the production period. The conditioncould be primary or secondary to another disease, such as Aleutian disease, mink virusenteritis, or distemper (Hunter, 1996b).
E. coli
is often isolated from outbreaks of diarrhoea in mink farms without furtherestimation of whether the disease should be considered as a primary or a superveningfactor. If
E. coli
is isolated from blood and internal organs, then it is assumed that
E. coli
has invaded its host through the digestive or respiratory tract and caused septicaemia. Inthe case of diarrhoea problems on the farm, the precise role of
E. coli
is not clear, becausethis organism is considered to be a part of the normal intestinal ¯ora in both healthy anddiseased animals. At least three explanations for the presence of
E. coli
in the gut of diarrhoeic mink are possible: (a) the
E. coli
isolated may be a part of the normal intestinal¯ora and the diarrhoea has another aetiology, (b) the mechanism responsible for infectioncreates an imbalance in the digestive system that enables
E. coli
to grow, and perhaps tocause secondary diarrhoea, or (c) the
E. coli
alone is responsible for causing diarrhoea.The latter is well known from humans, calves and pigs (Blood and Radostits, 1989). Suchbacteria possess speci®c virulence factors that determine attachment to intestinalsurfaces, invasion through intestinal mucosa or release of toxins that increase secretion of ¯uids to the intestinal lumen (Boedeker, 1984; Roth and Bolin, 1995). Some virulencefactors may be associated with certain O-groups, but, still, the de®nition of the O-groupalone is inadequate to predict the speci®c virulence of an
E. coli
isolate (Nataro andKaper, 1998; Roth and Bolin, 1995).At occurrence of clinical symptoms or sudden mortality related to
E. coli
infection on amink farm, treatment with antimicrobial agents will frequently be instituted prior to alaboratory examination. The drug chosen is not necessarily the most effective against theactual pathogen. When the laboratory results and the susceptibility pattern are available,the chosen drug can be evaluated and if necessary exchanged with another. Thisprocedure is inexpedient as the extended use of antimicrobials may result in developmentof resistance and ineffectiveness of the antimicrobial compounds used in mink production. It is well documented that
E. coli
may develop resistance to the mostcommonly used antimicrobial agents in veterinary practise. This has also beendemonstrated in an investigation of Danish mink farms in 1996, where the resistance
144
L. Vulfson et al./Veterinary Microbiology 79 (2001) 143±153
of
E. coli
isolates to tetracycline was 70% and to amoxycillin and ampicillin 60%(NùrregaÊrd and Kjems, 1999). So far, little research has been done on
E. coli
from mink,and to our knowledge, no investigations have been undertaken to study the role of
E. coli
as an aetiological agent of diarrhoea in mink. In the present study, 210
E. coli
strains wereisolated from diseased and healthy mink and subjected to serotyping and antimicrobialsusceptibility testing.
2. Materials and methods
2.1. Selection of farms
Six mink farms with outbreaks of diarrhoea were selected during the lactation andweaning period. The selection was made on the basis of examinations at the DanishVeterinary Laboratory. When
E.coli
in pure culture were isolated from carcassessubmitted to the laboratory, immediate action was taken to contact the farm for possibleinclusion in the study. It was subsequently decided to include the farm in the investigationon the basis of the following parameters: (a) anamnestic information: clinicalmanifestations on the farm and pathological ®ndings at autopsy should be indicative of
E. coli
infection, (b) antimicrobial treatment: no treatment should have been initiated inconnection with the outbreak prior to sampling, and (c) farm size should be representativefor Danish mink farms (800±3800 females). The participating farms were contacted andan initial visit to the farm was planned. During the visit, general information onmanagement and issues relating to the outbreak was collected. The farms were used as®xed effects in the multivariate logistic regression modelling and are described in thetables (Farms 1±6).
2.2. Sampling
On each farm, faecal cotton swab samples were collected from the rectum of 15 mink affected by diarrhoea and 15 apparently healthy mink. As supplementary material, deadanimals were collected from the selected farms. A second sampling from the sameanimals was made 2 weeks after the ®rst visit. If a tested animal died during this period, afaecal sample was instead collected from the mink in the neighbouring cage. The twovisits were used as ®xed effects in the multivariate logistic regression modelling (visit 1/ visit 2).
2.3. Bacteriological examination of specimens
The swabs were processed immediately after sampling on the farm by streaking ontoblood agar plates (Blood Agar base no. 2 Oxoid, CM271) supplemented with 5% calf blood and Drigalski agar plates and subsequently inserted into cryo-tubes containing VealInfusion broth (Difco) with 4% new born calf serum and 15% glycerol. Drigalski agar is amodi®ed MacConkey agar consisting of lactose 10 g, saccharose 10 g, sodiumthiosulphate
Â
5 hydrate 1 g, bromothymol blue 40 ml of a 2% solution, crystal violet
L. Vulfson et al./Veterinary Microbiology 79 (2001) 143±153
145
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