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Research Article
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Prevalence and Resistance to Antimicrobial Agents of Campylobacter
sp. Isolated from Dogs in India
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Amit Kumar Verma,
Amit Kumar,
Shanker Kumar Singh,
Anu Rahal,
Iftekhar Ahmed,
Deepti Singh,
Aashish Pratap Singh
and
Lalit Singh
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ABSTRACT
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Globally, Campylobacters have been reported as leading
cause of gastroenteritis in man as well as animals and considered as emerging
zoonotic problem particularly in developing countries including India. A cross-sectional
study was conducted to know the prevalence and epidemiological determinants
for Campylobacter spp. in dogs in and around Mathura city, Uttar Pradesh,
India. Based on isolation, cultural and biochemical characterization of bacteria,
the prevalence of Campylobacter spp. was 34.24%. Younger dogs (less than
1 year of age) were more likely to carry Campylobacter spp. High prevalence
of Campylobacter spp. supports the hypothesis that dogs, particularly
younger animals, may be an important source of Campylobacter infection
for humans. Breed-wise prevalence showed that non-descript dogs (45.97%) were
more likely to carry Campylobacter infections. Dogs showing clinical
signs of gastroenteritis were showing higher prevalence (47.21%) in comparison
to that of animals without gastro-intestinal disorders (15.04%). Out of 113
Campylobacter isolates of canine origin, two isolates were resistant
to all the nineteen antibiotics used in the study, while all the isolates were
resistant to Streptomycin, Ampicillin, Amoxycillin, Aztreonam, Lincomycin, Tetracyclin,
Oxytetracyclin and Penicillin. A high rate of resistance was observed to Cefotaxim
(97.35%), Peefloxacin (91.15%), Chloramphenicol (90.27%), Ofloxacin (84.07%),
Ciprofloxacin (83.18%), Cefaclor (80.53%), Nitrofurazone (76.11%), Norfloxacin
(74.33%), Gentamicin (42.48%), Amikacin (40.71%) and Enrofloxacin (36.28%).
Our results indicate Amikacin and Gentamicin as drugs suitable for the treatment
of campylobacteriosis in dogs.
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How
to cite this article:
Amit Kumar Verma, Amit Kumar, Shanker Kumar Singh, Anu Rahal, Iftekhar Ahmed, Deepti Singh, Aashish Pratap Singh and Lalit Singh, 2014. Prevalence and Resistance to Antimicrobial Agents of Campylobacter
sp. Isolated from Dogs in India. Journal of Biological Sciences, 14: 142-148. DOI: 10.3923/jbs.2014.142.148 URL: https://scialert.net/abstract/?doi=jbs.2014.142.148
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Received:
September 20, 2013; Accepted: October 29, 2013;
Published: January 11, 2014 |
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INTRODUCTION
Enteropathogenic bacteria are distributed worldwide and well to live as commensal
in the gastrointestinal tract of man as well as animals. The zoonotic potential
of these bacteria viz., Salmonella (Verma et al.,
2007, 2008, 2011a, b;
Sachan et al., 2013), E. coli (Malik
et al., 2013; Kumar et al., 2013)
and Campylobacter (Kumar et al., 2012a,
b) was very well established. Among these Campylobacters
are emerging food borne pathogens with increasing incidences and leads to severe
gastroenteritis and other complications like Guillain-Barre Syndrome, reactive
arthritis, haemolytic uraemic syndrome, meningitis, carditis, pancreatitis,
septic arthritis, etc., (Stern and Line, 2000; Tenkate
and Stafford, 2001; Hannu et al., 2002;
Skirrow, 2006; Levin, 2007;
Lindmark et al., 2009; Kirkpatrick
and Tribble, 2011; Man, 2011; Rajendran
et al., 2012; Bouwman et al., 2013;
Hauri et al., 2013). In animals, these
Campylobacters may cause diarrhoea and other reproductive disorders like
abortion, early embryonic mortality and many other complications (Humphrey
et al., 2007; Kumar et al., 2012a,
b).
Various studies have reported the association of Campylobacter species
with diarrhea in dogs (Steinhauserova et al., 2000;
Misawa et al., 2002; Kumar
et al., 2012a) however, their real role in canine enteritis is not
clear (Koene et al., 2004). Generally, children
are more susceptible to this infection (Wolfs et al.,
2001; Chattopadhyay et al., 2001). Although,
there is various reports from developed countries regarding prevalence of Campylobacters
in dogs but there is little published information about Campylobacter
infections and their drug resistance pattern in dogs of developing countries
including India. Therefore, the present study was undertaken to determine the
occurence of thermotolerant Campylobacter and their antibiogram in dogs
of Mathura city and nearby areas, Uttar Pradesh, India.
MATERIALS AND METHODS
Study design, area and sample collection: This cross-sectional study
was conducted during the period of almost one year in 2012-2013 in Mathura city
and nearby areas, Uttar Pradesh India (Fig. 1). Geographically,
the city is situated at the latitude 27°30N and longitude 77°40E
with an elevation of 174 metre above sea level. A total of 330 rectal swabs
for bacteriological culture were collected from dogs presented to Teaching Veterinary
Clinical Complex, Mathura with the epidemiological information viz., breed,
age, sex and health status.
Thermophilic Campylobacter isolation and identification: The
samples in the Campylobacter enrichment HiVeg TM broth base with addition
of polymixin B sulphate, rifampicin, trimethoprim and cycloheximide were incubated
at 42-43°C under microaerophilic conditions in the microaerophilic jars
with a lighting candle (Fig. 2). After incubation, the enriched
samples were properly shaken and sub-cultured onto Campylobacter selective
agar (HiMedia, Mumbai) supplemented with 10% defibrinated sheep blood and addition
of Polymixin B, vancomycin, trimethoprim and cephalothin for primary isolation
ofc thermophilic Campylobacter.
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Fig. 1: |
Map of Uttar Pradesh showing Mathura as study area |
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Fig. 2: |
Microaerophilic jar with a lighting candle |
The inoculated petridishes were incubated at 42°C for 48 h under microaerophilic
conditions. Suspected thermophilic Campylobacter colonies that were Gram
negative, curved, or spiral rods and showed corkscrew-like motion, were confirmed
biochemically (Skirrow and Benzamin, 1980; Garcia
et al., 1985).
Antibiogram: All the Campylobacter isolates were assessed for
their antimicrobial susceptibility testing using disc-diffusion method (Bauer
et al., 1966) following the NCCLS guidelines (NCCLS 2002). A total
of 19 antimicrobial discs (Hi-Media, Mumbai) of commonly used antibacterial
drugs viz., amikacin (30 μg), ampicillin (10 μg), amoxycillin (20
μg), Aztreonam (30 μg), Cefaclor (30 μg), Cefotaxim (30 μg),
ciprofloxacin (30 μg), chloramphenicol (30 μg), enrofolxacin (10 μg),
gentamicin (10 μg), Lincomycin (10 μg), Nitrofurazone (100 μg),
Norfloxacin (10 μg), Ofloxacin (5 μg), Oxytetracyclin (30 μg),
Peefloxacin (5 μg), Penicillin (10 units), streptomycin (10 μg) and
tetracyclin (30 μg) were used to assess the drug resistance pattern of
Campylobacter isolates.
RESULTS AND DISCUSSION
A total of 330 faecal samples were collected from dogs with and without clinical
signs of gastroenteritis and isolation of bacteria was attempted in all the
samples. Based on isolation, cultural and biochemical characterization of bacteria,
the prevalence of Campylobacter spp. was 34.24% (Table
1). The prevalence of Campylobacter spp. in faecal samples of dogs
is within the range (17-59%) as reported in various studies (Sandberg
et al., 2002; Engvall et al., 2003;
Koene et al., 2004; Rossi
et al., 2008; Parsons et al., 2010;
Kumar et al., 2012a, b).
Table 1: |
Occurrence of Campylobacter infection in dogs influenced
by age, sex, breed and health status |
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The variation between these studies might be either due to different study
population or methods used for detecting the bacteria (Guest
et al., 2007; Rossi et al., 2008;
Acke et al., 2009; Parsons
et al., 2010; Kumar et al., 2012a,
b).
Younger dogs (less than 1 year of age) were more likely to carry Campylobacter
spp. (Table 1). High prevalence of Campylobacter spp.
supports the hypothesis that dogs especially the puppies (less than 1 year of
age) may be an important source of Campylobacter infection for man. Our
findings were similar to previous studies (Engvall et
al., 2003; Acke et al., 2006, 2009;
Guest et al., 2007). But contrary to our findings,
a small number of studies conducted by Wieland et al.
(2005) and Tsai et al. (2007) have suggested
that age is not a predisposing factor for Campylobacter infection. Breed-wise
prevalence showed that non-descript dogs (45.97%) were more likely to carry
Campylobacter infections (Table 1). This might be due
to the way of living of non-descript dogs. As they roam outside the home freely
in comparison to other breeds of dog leading to more exposure and chances of
getting infection from stray dogs or animals in and around areas (Kumar
et al. 2012b).
Dogs showing clinical signs of gastroenteritis were showing higher prevalence
(47.21%) in comparison to that of animals without gastro-intestinal disorders
(15.04%) (Table 1). Similar to our study, various researchers
(Guest et al., 2007) reported the association
between Campylobacter infection and clinical signs, especially in younger
dogs (Fox et al., 1983; Nair
et al., 1985; Burnens et al., 1992).
Table 2: |
Drug susceptibility pattern of all the Campylobacter
isolates (n = 113) from dogs |
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However, contrary to our findings, there are various reports suggesting no
relationship between diarrhoea and Campylobacter spp. infection status
(Engvall et al., 2003; Acke
et al., 2006; Rossi et al., 2008;
Parsons et al., 2010; Kumar
et al., 2012b), suggesting that the organism is a commensal (Engvall
et al., 2003). Campylobacter infection were detected in 15.04%
of the dogs either apprarantly healthy or having some problem other than gastroenteritis
(Table 1) showing that these animals can be carriers of Campylobacter
species and may a source of infection for other pets and human beings (Acke
et al., 2006; 2009; Kumar
et al., 2012b).
All the Campylobacter isolates were tested for drug sensitivity using
19 antibacterial drugs and the results were shown in Table 2.
Out of 113 Campylobacter isolates of canine origin, two isolates were
resistant to all the nineteen antibiotics used in the study, while all the isolates
were resistant to Streptomycin, Ampicillin, Amoxycillin, Aztreonam, Lincomycin,
Tetracycline, Oxytetracycline and Penicillin. A high rate of resistance was
observed to Cefotaxim (97.35%), Peefloxacin (91.15%), Chloramphenicol (90.27%),
Ofloxacin (84.07%), Ciprofloxacin (83.18%), Cefaclor (80.53%), Nitrofurazone
(76.11%), Norfloxacin (74.33%), Gentamicin (42.48%), Amikacin (40.71%) and Enrofloxacin
(36.28%). Antibiotic sensitivity studies on Campylobacter isolates were
conducted by various researchers in different countries and reported varying
degree of resistance to same drug (Little et al.,
2008; Moran et al., 2009; Kumar
et al., 2012a). High resistance to ampi-cloxacillin, tetracycline,
ofloxacin, ciprofloxacin etc were previous reported by various researchers like
Little et al. (2008) and Miflin
et al. (2007). Quinolones were considered as suitable drug for treatment
of Campylobacteriosis (Uaboi-Egbenni et al., 2011)
but higher resistance to ciprofloxacin and ofloxacin was seen, which might be
either due to genetic mutations interfering with bacterial DNA gyrase (Greene
and Watson, 2003) or selection pressure due to injudicious use of antibiotics
(Norma et al., 2007; Biasi
et al., 2011). Present results indicated Amikacin and Gentamicin
as drugs suitable for the treatment of canine campylobacteriosis. This also
opens up therapeutic possibilities for these drugs in human medicine.
This cross-sectional study has the some limitations in its design; nevertheless,
it is performed in only little veterinary practice involving small number of
dogs and few related determinants. Therefore, there is a requirement of detailed
study considering larger population in order to establish a better understanding
of the epidemiology of Campylobacter infection in dogs of developing
countries like India and ultimately help in making strategies to control or
reduce the risk of this infection in man. The use of antibiotics as threauptic
and/or prophylaxis for man as well as animals should be monitored because acquisition
of antibiotic resistant strains of Campylobacters by man has serious
public health implications.
CONCLUSION
From the present study, it can be concluded that Campylobacter infection
was prevalent in dogs of study area suggesting their possible role in transmission
to human beings. Younger dogs (less than 1 year of age) were more likely to
carry Campylobacter spp. it is worth highlighting that dogs, particularly
puppies may be an important source of Campylobacter infection for humans.
High levels of resistance to antibiotics commonly used for prophylaxis and therapeutic
is of public health significance. Present results indicated Amikacin and Gentamicin
as drugs suitable for the treatment of canine campylobacteriosis. However, the
results emphasize the use of antibiotic sensitivity test be conducted before
prescribing the antibiotics.
ACKNOWLEDGMENTS
This study was supported b y Uttar Pradesh Pandit Deen Dayal Upadhayay Pashu
Chikitsa Vigyan Vishvidhyalaya Evum Go-Anusandhan Sansthan (DUVASU), Mathura,
India under university funding scheme. The authors are highly thankful to Head,
Department of Veterinary Microbiology and Immunology; Department of Veterinary
Epidemiology and Preventive Medicine, Director, Teaching Veterinary Clinical
Complex, Dean, College of Veterinary Sciences and Animal Husbandry, Director
Research and Honble Vice Chancellor, Uttar Pradesh Pandit Deen Dayal Upadhayay
Pashu Chikitsa Vigyan Vishvidhyalaya Evum Go-Anusandhan Sansthan (DUVASU), Mathura,
India; for providing all the necessary support and facilities for conducting
this study. We wish to thank staff of Department of Veterinary Microbiology
and Immunology especially Smt. Mamta for her technical assistance. The authors
are highly thankful to dog owners, who allowed their dogs to participate in
the study.
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