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Research Article
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Prevalence and Antibiogram of Campylobacter Infections in Dogs of Mathura, India |
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Rajesh Kumar,
A.K. Verma,
Amit Kumar,
Mukesh Srivastava
and
H.P. Lal
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ABSTRACT
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With the increasing trends of pet ownership the chances of campylobacteriosis are also increasing as these pets are kept in close visicinity of owners. The prevalence and antimicrobial sensitivity profiles of Campylobacter isolates from faeces of dogs attended in veterinary practice at Teaching Veterinary Clinical Complex, Mathura, India. During the period of investigation (October 2009 to April 2010), 100 rectal swabs from dogs were collected and transported to the laboratory for further investigations. Bacteriological examination revealed 51.00% prevalence rate of Campylobacter isolates in dogs faecal samples. The disc-diffusion method was used to know the susceptibility of all the 51 Campylobacter isolates against 10 commonly used antimicrobials in pet animal practice. High rates of resistance were observed to erythromycin (90.20%), tetracycline (88.23%), ampi-cloxacillin (88.23%), ciprofloxacin (80.39%), enrofloxacin (68.63%) and aymoxycillin-clavulanic acid (19.61%). All the Campylobacter isolates were susceptible to amikacin, levofloxacin and streptomycin. Erythromycin and ciprofloxacin are drugs for treatment of human campylobacteriosis. The high resistance rate to these drugs among Campylobacter isolates from dog faeces is of public health significance as dogs are supposed to be the main source of infection in human beings.
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Received: October 12, 2011;
Accepted: December 24, 2011;
Published: February 17, 2012
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INTRODUCTION
Enteropathogenic bacteria have long been recognized as a significant problem
owing to their pathogenicity potential to animals and their zoonotic risk to
humans. Among them, Campylobacters have been considered to be important
pathogens causing human gastroenteritis, arthritis, meningitis globally (Goldberg
and Rubin, 1988; Peterson, 1994; Baserisalehi
et al., 2006; Humphrey et al., 2007;
Frederick and Huda, 2011) and leading to serious impact
on public health (Ethelberg et al., 2004). Dogs
are contaminated by oral-fecal contact and by manifesting gastroenteritis or
acting as healthy carriers, serve as a potential source of infection to humans
(Bruce and Fleming, 1983; Goossen
et al., 1991; Burnens et al., 1992;
Ene et al., 1992; Moreno
et al., 1993; Torre and Tello, 1993; Fernandez
et al., 1994; Robinson and Pugh, 2002; Workman
et al., 2005; Sabry, 2009). Dogs in developing
countries like India often live in close proximity to humans (with the possibility
of direct transmission of pathogens) and have not been examined thoroughly for
Campylobacter sp. carriage. There is a dearth of information and research
on the prevalence and antimicrobial resistance studies of Campylobacters
in dogs in India in spite of the reports in Europe and other parts of the world
on dogs as a potential sources of infections for humans.
This study therefore, ascertained the prevalence and drug resistance profiles of Campylobacter spp. isolated from dogs in Mathura, India in order to provide updated information and the suspected role of dogs in its zoonotic significance. MATERIALS AND METHODS Rectal swabs were collected aseptically from 100 dogs presented to veterinary practice at Teaching Veterinary Clinical complex (University Veterinary Hospital), DUVASU, located in Mathura, India and transported at 4°C to the laboratory on the day of collection for the isolation of Campylobacter spp.
Isolation and identification of Campylobacter spp.: All the samples
were processed in Campylobacter Enrichment Hi Veg TM Broth Base (HiMedia,
Mumbai) with addition of polymixin B sulphate, rifampicin, trimethoprim and
cycloheximide (Campylobacter selective IV, HiMedia, Mumbai) and incubated
at 42-43°C under micro aerophilic conditions for 24 h. After incubation,
the inoculums was streaked onto selective media (Campylobacter selective
agar, HiMedia, Mumbai) supplemented with 10% defibrinated lysed sheep blood
and reconstituted contents of Campylobacter selective-I (HiMedia, Mumbai)
containing polymixin B, vancomycin, trimethoprim and cephalothin and incubated
for 48 h at 42-43°C under micro aerophilic conditions. Characteristic Campylobacter
colonies were picked up and subjected to presumptive identification like Grams
staining, motility, oxidase and catalase test and further subjected to biochemical
test for confirmation (Skirrow and Benzamin, 1980; Gracia
et al., 1985).
Antimicrobial sensitivity assay: All the Campylobacter isolates
were assessed for their antimicrobial susceptibility testing by the disc-diffusion
method following the NCCLS guidelines (NCCLS, 2002). The
following antimicrobial agents were used at the indicated concentrations (μg
disc-1 except where specified): using 10 commonly used antibiotic
discs (Hi-Media, Mumbai) viz., amikacin (30 g), amoxycillin-clavulanic acid
(20/10 μg), ampicloxacillin (10 μg), ciprofloxacin (30 μg), chloramphenicol
(30 μg), enrofolxacin (10 μg), erythyromycin (15 μg), levofloxacin
(5 μg), streptomycin (10 μg) and tetracyclin (30 μg).
RESULTS Campylobacter spp. was isolated from 51 dogs of the total 100 dogs (percent positivity 51.00%). The results of antibiotic sensitivity test for the ten antimicrobial agents for Campylobacter spp. is shown in Table 1. Using the disc diffusion method, 46 out of 51 isolates of Campylobacter (90.20%) were resistant to erythromycin, 45 to tetracycline (88.23%), 45 to ampi-cloxacillin (88.23%), 41 to ciprofloxacin (80.39%), 35 to enrofloxacin (68.63%) and 10 to amoxycillin-clavulanic acid (19.61%). All the Campylobacter isolates were susceptible to amikacin, choramphenicol, levofloxacin and streptomycin.
Table 1: |
Antibiotic sensitivity test of Campylobacter spp.
isolates of dogs |
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S: Sensitivity, I: Intermediate, R: Resistant |
DISCUSSION
Elucidating the shedding patterns and prevalence of Campylobacters in
the faeces of dogs is a prerequisite for effective healthcare strategy against
zoonotic infections. The species distribution of Campylobacter isolates
from dogs and other animals differs considerably between publications and years
(Hald and Madsen, 1997; Lopez et
al., 2002; Sandberg et al., 2002; Hald
et al., 2004; Workman et al., 2005;
Mohammad and Mohagheghi, 2006; Baserisalehi
et al., 2007b; Huat et al., 2010).
The introduction of antimicrobial agents in human and animal therapy has had
a great impact on population. The first agents were introduced during the 1930s
and resistance to these drugs gradually emerged with their worldwide use. Campylobacter
spp. is classified as an emerging human pathogen and recently, concern regarding
the prevalence of campylobacteriosis has increased because of the frequent isolation
of antimicrobial-resistant strains from humans and animals. After exposure to
ten antibiotics, several isolates showed multiple resistances to most of the
antibiotics used. Studies related to the sensitivity to antibiotics of Campylobacter
spp. in different countries show different degrees of resistance to the same
drug (Guevremont et al., 2006; Han
et al., 2007; Little et al., 2008;
Moran et al., 2009). In this study, majority
of Campylobacter spp. isolates showed resistance to at least 5 of the
antibiotics tested, indicating multi-drug resistance. In the present work, all
isolates were sensitive to amikacin, chloramphenicol, levofloxacin and streptomycin.
Sensitivity to chloramphenicol by all Campylobacter isolates was also
observed in pigs (Saenz et al., 2000; Guevremont
et al., 2006), whereas, similar resistance patterns for Campylobacter
spp. was obtained from humans (Bardon et al., 2009)
and chickens (Miflin et al., 2007).
Majority of the isolates were resistant to erythromycin, tetracycline, ampi-cloxacin,
ciprofloxacin and enrofloxacin. Resistance to tetracycline by Campylobacter
isolates from humans, dogs and other animals may range from 15 to 94% (Modolo
et al., 1991; Gaudreau and Gilbert, 1998;
Saenz et al., 2000; De Vega
et al., 2005). The great variability in this antibiotic's efficacy
is probably due to its worldwide use in cattle, both at therapeutic or low doses;
this would increase selective pressure on bacteria. Clinical assays, however,
have shown the therapeutic efficacy of tetracycline in treating dogs with Campylobacteriosis
and a decrease in re-excretion rate (Abrahans et al.,
1990; Burnens et al., 1992). High resistance
to ampi-cloxacillin for Campylobacter spp. i.e., 57.3% (Little
et al., 2008) and 65.7% (Saenz et al.,
2000) was also recorded in samples from pigs and 43.1% (Han
et al., 2007) and 40.8% (Miflin et al.,
2007) in samples from chicken. Similar results were obtained in this study,
where 88.23% of the strains showed resistance to ampi-cloxacillin.
Campylobacter was frequently sensitive to quinolones; however, an increased
resistance to these drugs is seen, probably due to genetic mutations interfering
with bacterial DNA girase (Greene and Watson, 2003).
Selective pressure caused by the indiscriminate use of these drugs in aviculture
is a contributory factor. Previous studies (Saenz et
al., 2000; Norma et al., 2007; Biasi
et al., 2011) reported the greatest resistance of Campylobacter isolates
to quinolones among various antibiotics similar to results obtained in the current
study, in which 80.39% of the isolates were resistant to ciprofloxacin. Contrary
to our findings, sensitivity to ciprofloxacin for all the 152 strains of Campylobacter
spp. isolated from chicken in Australia (Miflin et al.,
2007); 70 isolates from domestic animals and poultry from India (Baserisalehi
et al., 2007a); isolates from environmental samples (Baserisalehi
and Bahador, 2008) was also reported earlier, whereas ciprofloxacin resistance
in only 0.3% of isolates was confirmed from cattle in Canada (Inglis
et al., 2005). The resistance patterns displayed by Campylobacter
isolates from dogs to fluoroquinolone (ciprofloxacin) and macrolides (erythromycin)
classified as second line and first line antimicrobials are of particular importance,
since patients suffering from Campylobacteriosis are usually treated with these
antimicrobials agents (Uaboi-Egbenni et al., 2011).
Recent scientific studies has shown that Campylobacter antimicrobial
resistance can be related to some specific genes and the dissemination of these
genes of microorganisms to their progeny and across to other unrelated bacteria
species through extrachromosomal DNA fragment called the plasmid from one animal
species to another and to humans is possible (Baserisalehi
and Bahador, 2008; Apata, 2009). Antimicrobial resistance
observed in the present work might be due to the indiscriminate and irrational
use of antimicrobials (Tambekar et al., 2007)
in animals for preventive or therapeutic purposes irrespective of etiological
agents. Given the relevance of the genus Campylobacter in human gastroenteritis,
its occurrence in companion animals such as dog and the fact that majority of
Campylobacters showed multi-drug resistance, a continuous surveillance
and monitoring of the prevalence and the antimicrobial resistance of Campylobacter
spp. in dogs and other pet animals is essential to the implementation of effective
policies for controlling and preventing contamination and infection by this
pathogen. The use of antibiotics as threauptic and prophylaxis for animals should
be carefully evaluated and monitored because acquisition of antibiotic resistant
strains of Campylobacters by man has serious health implications.
CONCLUSION In view of the heterogeneity in the results reported in the literature in comparison to data in this study, we recommend that Campylobacter antimicrobial susceptibility tests be performed for therapeutic purposes with the strict hygienic measures to prevent transmission from pets to owner. Our results indicate amikacin, chloramphenicol, streptomycin and levofloxacin as drugs suitable for the treatment of canine Campylobacteriosis. This also opens up therapeutic possibilities for these drugs in human Campylobacteriosis. ACKNOWLEDGMENT
The authors of this study are highly thankful to Dr. Manoj Gupta, Incharge,
department of Veterinary Microbiology and Immunology for help in conducting
the laboratory works; Dean, Veterinary faculty and Honble Vice Chancellor,
DUVASU, Mathura; for providing funds and necessary facilities for conducting
the study; the staff at the TVCC; for collecting the samples; staff at department
of veterinary microbiology and immunology for assistance in laboratory works
and the owners of the dogs who gave their permission for their dogs to take
part in the study.
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