Prevalence , seasonality and antibiotic susceptibility of thermophilic Campylobacters in ceca and carcasses of poultry birds in the “ live-bird market ”

Thermophilic Campylobacter spp. are the primary cause of human diarrhea. The common source of infection is contaminated poultry. This study aimed to establish the prevalence of Campylobacter spp. in poultry obtained from ‘pluck shops’ and provide a baseline of resistance profiles of Campylobacter spp. isolates obtained. A biphasic approach, with qualitative detection as well as quantitative enumeration of Campylobacter spp. was used. We examined 240 samples each, of carcasses and cecum of poultry obtained from ‘pluck shops. Amongst the cecum samples and carcasses, 59.5 and 57%, respectively were positive for Campylobacter spp. The average Campylobacter spp. concentration was 2.69 (S.D 0.419) log10 CFU/mL and 4.55 (S.D 0.607) log10 CFU/g for carcass rinsate and cecum, respectively. Of the 225 Campylobacter isolates studied, 76.9% were identified as Campylobacter jejuni and 23.1% as Campylobacter coli. Susceptibilities for 112 strains of C. jejuni and 31 strains of C. coli were determined for 12 antibiotics by the agar diffusion technique. According to the minimal inhibitory concentration, a marked resistance to gentamycin and chloramphenicol was also demonstrated. According to the antibiotic resistance profiles, the isolates appeared to differ from each other.


INTRODUCTION
Campylobacter jejuni and Campylobacter coli are commonly associated with poultry as well as poultry products because they are commensal of the avian gut.Campylobacter spp. is the causative agent responsible for human Campylobacteriosis (Humphrey et al., 2007).The central aspect of Campylobacter infection in humans is an acute inflammatory gastroenteritis.The disease begins with a battery of symptoms, which may or may not be mutually exclusive.Abdominal cramps, fever, rigors, dizziness, headache, convulsions, delirium, nausea and even myalgia may be among the prodrome preceding copious, runny, watery and bile stained diarrhea.Poultry meat is the principal risk factor associated with Campylobacter infections in man (Kapperud et al., 1992;Hudson et al., 1999;Allos, 2001).The different serious sequels such as Guillain-Barré´ syndrome (GBS) or Reiter syndrome are characterized by polyneuritis of the peripheral nerves and may be seen in 1 of 1000 patients infected with C. jejuni.These sequelae may be associated with paralysis and severe neurologic deficits (Nachamkin et al., 1998;Kuroki *Corresponding author.E-mail: bpkapadnis@yahoo.com.et al., 1993).These bacteria are associated with comercially produced chickens from free range farms as well as mass-produced broilers and laying hens (Rodenburg et al., 2004;Miraglia et al., 2007).
Campylobacter sp.contamination commences at the farm level (Byrd et al., 1998).Colonization levels from log 5 to log 9 CFU/g of cecal contents have been reported in broilers (Berndtson et al., 1996).Horizontal transfer of Campylobacter ensures the spread of the organism within the whole flock in a few days (Gerdemann, 1996).It has been proposed that human infections follow increased Campylobacter in animals (Baker et al., 2012).
In India, the broiler poultry is processed and vended as a fresh product to consumers from a particular location defined as a "pluck shop" (Rodrigo et al., 2005).
The poultry industry thrives solely as a 'live-bird' market comprising the pluck shops.Traditional poultry facilities at the wholesale or retail level are manual with negligible sanitary practices taken on the flooring or by the personnel.Only scarce 5% of all poultry meat in India is processed mechanically in industries under hygienic conditions using recommended equipment whereas most poultry meat enters the food chain via the 'live-bird' market through 'pluck-shops' (Reardon and Gulati, 2008).This may be because consumers prefer fresh meat.The fact that mechanical slaughtering process incorporating equipments may reduce the level of contamination by 100 to 1000 times (Rosenquist et al., 2006), is usually overlooked due to cultural disposition to assume that hand slaughtered birds are the fresher and healthier ones.
An infected carcass indicates an infected bird.This is because unlike most other bacteria implicated in food poisoning, Campylobacter are fastidious and cannot multiply outside their animal host.Hence, any contamination on the carcass is from within the birds itself.The Campylobacter levels on the carcass represent an important source of consumer exposure and potential risk for infection (Stern and Robach, 2003).Thus, the quantification of the Campylobacter contamination levels in the birds available through 'live-market' is predominantly imperative.However, Indian reports on human Campylobacteriosis or even incidence in poultry are sparse.The above fact has underlying global consequences in light of the increasing numbers of travelers entering and poultry exports exiting the Indian subcontinent.
Campylobacter colonization in poultry has been reported to follow a seasonal pattern, peaking in the warmer months (Jacobs-Reitsma et al., 1995;Boysen et al., 2011;Nylen et al., 2002;Hudson et al., 1999).To date, there have been very few studies investigating the prevalence and seasonality of Campylobacter spp. in the Indian poultry industry.In the present study, we have quantified the Campylobacter contamination levels on carcasses in the 'live-market 'and identified the various thermophilic Campylobacter found.Additionally, we examined the antibiotic susceptibility of these isolates.To our knowledge, this is the first comprehensive report exploring the seasonality of Campylobacter in poultry from tropical environments.

Determination of sample size
The sample size was calculated using the formula proposed by Thrusfield, for a large (theoretically 'infinite') population on assuming simple random sampling with annual expected prevalence (p) of 50%, desired confidence level (Z) of 95%, corresponding to a Zα value of 1.96 (Thrusfield, 1995) and using the formula: The population size is infinite hence; the sample size is 384 with accuracy (L) of 5% and sample size of 196 with accuracy (L) of 10%.Considering practicality of the sampling and the time taken for processing, 240 (L= 6.3%) carcasses were sampled over the course of one year (May 2008 to April 2009) from vendors located at four regions within Pune viz.Aundh, Camp, Chinchwad and Hadapsar.

Qualitative detection of Campylobacter spp.
The qualitative detection of Campylobacter was carried out to understand its prevalence and find the percentage of samples positive carrying Campylobacter.Presence or absence testing was performed on 240 carcass and cecum samples each according to modifications of ISO 10272-1:2006(E).Briefly, the method was as follows: 25 g of carcass meat sample was placed into enrichment broth (1:10), homogenized and then incubated at 42 ± 1°C for 48 h under the microaerobic conditions (5% O 2 , 10% CO 2 , 85% N 2 ).Sterile Preston broth (HiMedia, India, M899) supplemented with selective supplement (HiMedia, India FD-042) containing polymyxin B (5 IU/mL), rifampicin (10 µg/mL), trimethoprim 10 (µg/mL) and cycloheximide (100 µg/mL) along with 10% horse blood (Haffkine Biopharmaceutical Ltd, Pune) was used as enrichment broth.Then one loopful of enrichment broth was streaked onto mCCDA agar plates (ISO, 10272-1; Stoyanchev et al., 2007;Habib et al., 2011).For qualitative analysis, about 0.5 g of caecal contents were directly plated on mCCDA plates (Hansson et al., 2010) Both sets of mCCDA plates were then incubated at 42 ± 1°C for 24 h under microaerobic conditions using McIntosh and Filde's anaerobic jar (Hi-Media, IndiaAnaerobic System Mark VI, LE013).Plates were inspected to detect the presence of colonies presumed because of their characteristics of Campylobacter.

Quantitative determination to enumerate Campylobacter counts
For quantification of Campylobacter, modification of the ISO 10272-2:2006(E) was used.Briefly, 48 whole carcasses were rinsed in a large plastic bag containing 500 mL of sterile buffered peptone water to obtain carcass-associated microflora.Decimal dilutions in buffered peptone water were prepared from 1 mL of rinsate.One hundred microlitres of each dilution was spread plated in duplicate onto mCCDA (HiMedia, India, FD-042).Then forty-eight ceca were obtained, 12 from each region.Cecal contents were weighed and diluted in buffered peptone water.Then a 10-fold serial dilution in peptone water was prepared.One milliliter from each dilution was then plated on mCCDA plates (Hansson et al., 2010).

Microbiology of presumptive isolates
The plates were examined for round, translucent, raised, convex colonies with entire edge, and a tendency to spread indicating motile nature.The suspected colonies were contradistinguished by the presence of slender, spiral, curved and Gram-negative rods with typical corkscrew, darting motility under hanging drop examination.Presumptive colonies were oxidase positive and unable to grow under aerobic conditions, when incubated on fresh mCCDA plates at 37°C.Presumptive colonies, based on colony morphology, were restreaked on Muller-Hinton-based blood agar plates (Hi-Media India , M-173) supplemented with 10% (v/v) horse blood and incubated microaerobically at 42°C for 24 h.Isolated colonies were then restreaked for purity on mCCDA and incubated microaerobically at 42°C overnight.Biochemical tests, which consisted of hippurate hydrolysis, catalase test, indoxyl acetate hydrolysis and H 2 S test, were performed on colonies isolated from the blood agar plates.The HiCampylobacter™ Latex Test Kit (Hi-Media, India), a rapid latex agglutination test was used for confirmation of the isolates as thermophilic Campylobacters.
For 16S rRNA, tubes were subjected to 35 cycles of 94°C for 30 s, 55°C for 30 s and 72°C for 1 min, followed by a 7-min extension at 72°C.The sequencing reaction for either PCR reaction was performed in 25 μl volumes containing 1 μl DNA; 20 mM Tris-HCl (pH 8.3); 50 mM KCl; 2.5 mM MgCl 2 ; 200 μM each dNTP; 0.4 μM of each primer; and 0.625 units of Taq DNA polymerase.
Sequencing was performed using degenerate primers 907R (Sigma).Sequence data were obtained using a 3730 DNA Analyzer (Applied Biosystems, Foster City, CA).The sequence analysis software used was ChromasPro v1.34.Identity of the isolates was confirmed by BLAST on NCBI.

Antibiotic susceptibility
The minimum inhibitory concentration was determined for 143 selected isolates (112 C. jejuni, 31 C. coli) of Campylobacter which showed resistance towards at least one antibiotic.The agar dilution method recommended by the Clinical and Laboratory Standards Institute (CLSI) (previously National Committee for Clinical Laboratory Standards, NCCLS), subcommittee on Veterinary Antimicrobial Susceptibility Testing (NCCLS, 1999) was used as a standard.The tested antimicrobial agents (HiMedia, India) were as follows: ampicillin, azithromycin, chloramphenicol, ciprofloxacin, doxycycline, gentamycin, nalidixic acid, norfloxacin and tetracycline.There is a dearth of internationally validated criteria for breakpoints of susceptible or resistant isolates for Campylobacter.Conse-quently, where breakpoints from CLSI were not available, esta-blished breakpoints were used (Luber et al. 2003a;Yabe et al., 2010;Gu et al., 2009;Luangtongkum et al., 2007).Suspensions of isolates were prepared and were adjusted to a turbidity equivalent to a 0.5 McFarland standard, which is equivalent to 1.5 x 10 8 CFU/mL.This was diluted to 1:100 leading to a concentration of 1.5 x 10 6 CFU/mL.The final inoculum on the agar was approximately 3 x 10 3 CFU/spot.These suspensions were inoculated onto Mueller-Hinton agar (HiMedia, India) containing the twofold dilution series of antibiotics and supplemented with 5% defibrinated horse blood (Luangtongkum et al., 2007).Plates were incubated at 42°C for 48 h, in a microaerophilic atmosphere.Growth was assessed after incubation and the MIC value was determined to be dilution of the antibiotic, which inhibits the growth of the isolate under study.Antimicrobial susceptibility was determined for concentrations ranging from 0.125 to 256 µg/mL.To ensure reproducibility, MIC determinations were repeated at least thrice.

Statistical analysis
Campylobacter counts were converted to a logarithmic scale to approximate the results to normal distribution.Campylobacter detection was recorded in binary variables in terms of Campylobacter presence or absence.Enumeration results were recorded as CFU/mL of rinse liquid for carcasses and CFU/g of cecal contents.Campylobacter mean counts (carcass and ceca) were compared between months and between seasons.P ≤ 0.05 was considered statistically significant.Statistical analysis of the data was carried out using Minitab (Version 14).

Overview of Campylobacter contamination
Table 1 shows prevalence of Campylobacter spp. in chicken carcasses and intestine from broilers at pluck shops.All the live processing units were found to be positive for Campylobacter.More than half (59.6%) of the cecal samples and 57% of the carcasses tested were positive for Campylobacter spp.The prevalence of Campylobacter spp. in contaminated carcasses was highest in the Aundh area (65%), and lowest in the Camp region (48.3%).The prevalence of Campylobacter contaminated ceca was highest in the Camp region (66.7%) and lowest in Hadapsar (55%).Of the 225 isolates studied, 76.9% (173) were identified as C. jejuni and 23.1% ( 52) C. coli.The isolates were positively identified using the PCR based assays for identification of Campylobacter spp.Since the biochemical analysis revealed the presence of only C. jejuni and C. coli amongst the isolates, a single PCR assay that amplified a part of the flagellin gene was deemed sufficient for the study.The PCR yielded the expected amplicon of product size of 450 bp, with the primers specific for flagellin gene.Further, the PCR assay confirmed that the isolates obtained belonged to Campylobacter spp.On amplification of 16SrRNA gene and carrying out BLAST, along with differentiation based on biochemical tests, the isolates were identified as C. jejuni or C. coli.
The count data for carcass contamination showed that 81.7% of the samples were contaminated with 10 2 to 10 3 CFU/mL, while 16.7% of the samples showed contamina- tion greater than 10 3 CFU/mL.The average Campylobacter concentration was 2.69 log 10 CFU/mL, with a standard deviation of 0.41 log 10 CFU/mL.The count data for cecal samples showed that 14.6% of the samples were contaminated with 10 3 to 10 4 CFU/g, while 58.3% of the samples showed contamination between 10 4 and 10 5 CFU/g and 27% greater than 10 5 CFU/g.The average Campylobacter concentration was 4.55 log 10 CFU/g, with a standard deviation of 0.60 log 10 CFU/g.

Qualitative study of prevalence
This entails the percentage of samples positive for Campylobacter. Figure 1A and B show monthly occurrence of Campylobacter spp. in chicken intestine and carcasses, respectively.In the 12 months, highest and lowest poultry prevalence of Campylobacter was seen in May and January, respectively among the carcass samples, whereas the cecal samples showed highest and lowest prevalence in June and December, respectively.Prevalence studies show that highest prevalence in cecum as well as carcasses was noted in monsoon.Lowest prevalence in cecal and carcasses was seen in post-monsoon and winter, respectively.

Quantitative enumeration
This entails the Campylobacter load and thus the numbers on the carcass.Figure 2A and B show seasonal occurrence of Campylobacter spp. in chicken intestine and carcasses, respectively.Highest and lowest numbers of Campylobacter were seen in October and December, respectively among the carcass samples, whereas the cecal samples showed highest and lowest numbers of Campylobacter in May and September, respectively.The highest and lowest numbers of Campylobacter in poultry ceca were seen in the post-monsoon and monsoon season, whereas the highest and lowest numbers of Campylobacter in poultry carcasses were seen in summer and winter, respectively.

Antimicrobial resistance of the Campylobacter isolates
The prevalence of antimicrobial resistance patterns in the C. jejuni and C. coli isolates are presented in Tables 2  and 3, respectively.Amongst all the C. jejuni and C. coli isolates, 64.7 and 59.6% respectively were resistant to one or more antibiotics.35.9 and 25% of C. jejuni and C. coli isolates respectively showed multidrug resistance to 4 or more antibiotics.
The MIC was determined for only those isolates, which showed resistance to one or more antibiotic.Minimal inhibitory concentrations of 12 antimicrobial agents were determined via agar dilution for 112 isolates of C. jejuni and 31 isolates of C. coli (Table 4).The MIC 90 s for C. jejuni were 64 μg/mL for ciprofloxacin and nalidixic acid, 32 μg/mL for ampicillin, erythromycin, gentamycin, norfloxacin and tetracycline, 4 μg/mL for streptomycin, 2 μg/mL for chloramphenicol and doxycycline, 0.5 μ g/mL for clindamycin and ≥ 0.0625 μg/mL for azithromycin.The MIC 90 s for C. coli were 64 μg/mL for ciprofloxacin and chloramphenicol, 32 μg/mL for ampicillin, nalidixic acid, norfloxacin and tetracycline, 16 μg/mL for gentamycin, 4 μg/mL for clindamycin, erythromycin and streptomycin and 1 μg/mL for azithromycin.
Gentamycin, erythromycin, ciprofloxacin, norfloxacin and tetracycline resistance was common amongst the isolates.The C. jejuni isolates had norfloxacin MICs as high as 256 µg/mL, indicating a high-level resistance to fluoroquinolones.Amongst quinolones (ciprofloxacin, norfloxacin and nalidixic acid), tetracyclines (tetracycline and doxycycline), macrolides (erythromycin and azithromycin) and clindamycin, the overall resistance rates were statistically similar between C. jejuni and C. coli.The MIC values however in C. jejuni were higher than in C. coli.As compared to other antimicrobial agents, the resistance rates to ampicilin were significantly different in C. jejuni and C. coli with the C. coli isolates showing higher prevalence of ampicillin resistance (48%).The overall prevalence of azithromycin resistance was low in C. jejuni (6.3%) and C. coli (5.7%).Another notable observation of this study was erythromycin resistance, which was moderate among the C. jejuni isolates (26%), whereas not a single case of erythromycin resistance was seen amongst the C. coli isolates.

Prevalence of Campylobacter in retail poultry meat from "pluck-shops"
In countries where frozen or chilled poultry is predominantly consumed, rates of isolation of thermophilic Campylobacters from carcasses were: 49.5% in Spain (Domínguez et al., 2002), 35.2% in Bulgaria (Stoyanchev et al., 2007), 44% Germany (Näther et al., 2009) and 52.5% in the US (Son et al., 2007).The rates of isolation of Campylobacter contamination are higher in countries with traditional "pluck shop" based wet markets: 63% in Iran (Taremi et al., 2006), 68.3% in Korea (Han et al., 2007) and 83.9% in Trinidad.Studies in Malaysia have shown the overall rate of contamination for Campylobacter in modern processing plants and in traditional wet markets were 61.1 and 85.6%, respectively (Rejab et al., 2012).The prevalence of Campylobacter-positive carcasses, found in this study, was observed to be comparable to prevalence reports for chicken carcasses from other countries were poultry wet-markets is common (Rodrigo et al., 2005).Previous studies in India also concur with our findings with 39.3% of the tested poultry positive for Campylobacter in Calcutta (Chattopadhyay et al., 2001), 64% in Vellore (Rajendran et al., 2012) and 17.14% in the Meghalaya-Assam region (Rizal et al., 2010).Our findings on the prevalence of Campylobacter, based on cecal samples, corresponds well with those reported by Stern and Robach (2003) with an average population of 4.6 log 10 CFU/g in 1995 and an average population of 5.17 log 10 CFU/g in 2001 (Stern and Robach, 2003).These
We postulated that since the numbers of this organism in the cecal contents correspond with those observed in other countries, the higher numbers of contaminated carcasses as well as higher Campylobacter loads can possibly be due to lack of industrial processing.Industrial poultry processing comprises of killing, scalding, defeathering and evisceration of birds mechanically.
Each step is followed by washing with chlorinated water.Finally, the processing is culminated with a rapid cooling on carcass chillers.The number of Campylobacter sps.microorganisms sequentially reduce at each step, followed by a final wash with chlorinated water and processing aids that almost finishes the contamination, leading to a low count on the end product (Keener et al., 2004).The mechanical slaughtering reduces Campylobacter contamination by a 100 to 1000 fold (Rosenquist et al., 2006).
These reasons may explain the high level of contamination found in poultry carcasses from India.Manual slaughtering and evisceration by hand leading to rupture of intestinal viscera may lead to fecal contamination of carcasses.In addition, the lack of washing steps with water or processing aids may be responsible for the increased numbers of Campylobacters observed in Indian poultry.

Seasonality
We are not confirming a definite seasonality in the Campylobacter numbers or its prevalence in birds.However, noticeable peaks are seen during the warmer months.Consequently, we support this concept of seasonality due to the presence of dual peaks, one in the month of May and the second in October.Both peaks occur during warmer months.Earlier studies on seasonality of Campylobacter sp. by Singh and colleagues (2008) showed that the highest prevalence of C. jejuni in faecal samples was reported during rainy season.This is in agreement with our results.However, among the different months, the highest prevalence was found in September, whereas our study shows highest prevalence was seen in October.These findings can be explained by the regional differences in temperature.In addition, higher isolation rates of Campylobacter spp.during the summer and monsoon months in children has been reported in Calcutta (Bhadra et al., 1992).
The seasonal structure seen in India is very different from that of European countries with an absence of the spring and autumn seasons and presence of monsoon and the post monsoon seasons.Even so, higher recovery rates were reported during the warmer months of the year in the U.S (Willis and Murray, 1997) as well as India (Singh et al., 2008).Seasonality in Campylobacter  colonization of poultry has been studied (Berndtson et al., 1996).Seasonal peak in humans coincides with a peak in poultry isolates (Meldrum et al., 2005).The Campylobacter population in the intestines of cattle, lamb and poultry differ considerably with changing seasons (Wallace et al., 1997;Stanley et al., 1998a, b).Hence it is deducible that carcass contamination potentially varies with the season (Jones, 2001).Despite these pressing arguments, seasonality is not often studied with reference to Campylobacter spp.

Number of isolates of C. coli
Studies on packed meats are not conducive to understanding seasonal variations because of the longer shelf life of these products.Frozen or chilled poultry is stored by companies and released as per demand and may be further stored by retailers before the actual sale.Furthermore, time lapses occur during poultry processing, transportation and retail storage.Studies from "pluck-shops" make an interesting model because birds are sold immediately after culling.

Antibiotic resistance
The resistance rates of Campylobacter to each antimicrobial agent differ considerably in several countries (Payot et al., 2004;Luberet al., 2003b;Aquino et al., 2002;Gupta et al., 2004).In our data set, the susceptibility of Campylobacter isolates has been evaluated by using the minimal inhibitory concentration method, therefore  enabling the determination of precise concentration at which the microorganism failed to grow.Campylobacter enteritis are usually treated using fluoroquinolones and macrolides (Allos, 2001).On the other hand, recent studies have reported the appearance of fluoroquinolones resistant Campylobacter spp.among poultry flocks (Niwa et al., 2001), necessitating the survey of prevalence of Campylobacter spp. in poultry and their antimicrobial resistances.Earlier reports from India show 30.6% of strains were multidrug resistant (Jain et al., 2005).This is in agreement with our result where 35.83 and 25% of C. jejuni and C. coli isolates respectively showed multidrug resistance to four or more antibiotics.
C. coli strains are reported to have tendency to acquire resistance to macrolides, specifically erythromycin.In contrast, C. jejuni isolates remain sensitive to erythromycin.C. coli has frequently been found to be resistant to erythromycin and other macrolides (Nayak et al., 2005;Kim et al., 2006).This propensity was not observed is our data set.Gentamycin resistance has been reported to be absent in studies from Europe and U.S (Hariharan et al., 2009).However, data from Asian countries suggests otherwise.The findings in our study are in accordance with the data obtained from other Asian countries (Chen et al., 2010).Earlier studies from this region have indicated that the frequency of antibiotic resistance is high in Campylobacters (Baserisalehi et al., 2005).In 2011, a new policy on containment of antimicrobial resistance  was enacted in India.It will be interesting to note any change of antimicrobial resistance patterns in future surveys of Campylobacter isolates in this scenario.
There is a paucity of information on the levels of Campylobacter and its prevalence in poultry carcasses from "pluck-shops" or wet poultry markets.Data emerging from western countries is usually from chicken carcasses picked at the end of stringent mechanical poultry processing.However, this is not always the case in many Asian countries.Assuring good manufacturing along with food safety practices is crucial.The manual and laborious method of poultry processing used in India needs to be streamlined and regulated.The cultural confines, which endorse the con-sumption of freshly culled birds, may or may not change.Hence, a more hygiene oriented, education based approach needs to be taken and poultry processors or "pluck-shop" owners need to be educated on a more informed protocol of processing.Minimum hygiene standards must be set and adherence to the same must be regulated.
This research shows that higher mean concentrations in the cecum have been seen in warmer months.Earlier research postulates that low intestinal Campylobacter concentration leads to low fecal contamination of the carcasses during the slaughter steps and conversely with high concentrations (Rosenquist et al., 2006).It has been proposed that the probability of meat contamination increases when the prevalence of Campylobacters is high within the flock and higher numbers of Campylobacters are present in the intestines (Nauta et al., 2009).Due consideration must be given to these factors and a real time surveillance system must be brought into practice.
Since Campylobacter sp. is considered a major pathogen associated with food borne disease worldwide, this study revealed the prevalence of antimicrobial resistance in Campylobacter sp. from poultry, which is the most consumed animal food in India.Surveillance systems must be brought into place to monitor the use of antibiotics in poultry.The results of this study suggest that timely scrutiny of the presence of Campylobacter sp.needs to be conducted and reported in poultry to reduce their numbers thereby preventing cases of

Figure 1 .
Figure 1.Monthly occurrence of Campylobacter in chicken intestine (A) and in carcasses (B) from May 2007 to April 2008.▲, Percentage of positive samples; lined columns, Microbial counts for cecca and carcasses expressed as log cfu/g and log cfu/ml, respectively.

Figure 2 .
Figure 2. Seasonal occurrence of Campylobacter in chicken intestine (A) and in carcasses (B).▲, Percentage of positive samples; lined columns, Microbial counts for cecca and carcasses expressed as log cfu/g and log cfu/ml, respectively.

Table 1 .
Prevalence of Campylobacter in chicken carcasses and intestine from broilers at pluck shops.

) of samples positive for Campylobacter Mean microbial count Intestine c Carcass c Cecca d Carcass d
Total number of samples tested was 48, that is, 12 from each region.Microbial counts for cecca and carcasses are expressed as log cfu/g and log cfu/ml, respectively. d

Table 2 .
Drug resistance patterns in Campylobacter isolates from raw chicken in India, during the period of May 2008 to April 2009.

Table 3 .
Multi drug resistance patterns in Campylobacter isolates from raw chicken in India, during the period ofMay,  2008 to April 2009.

Table 4 .
Distributions of MICs of 12 antimicrobial agents for 112 C. jejuni isolates and 31 C. coli isolates from "pluck-shop" broiler chicken.