Isolation, molecular identification and antimicrobial resistance patterns of Campylobacter species of dairy origin: First report from Bangladesh

Thermophilic Campylobacter species are common cause of animal and human bacterial diseases with growing resistance to antimicrobials. The aim of this study was to determine the prevalence and antimicrobial susceptibility pattern of Campylobacter species from bovine, knives and personnel in Jimma Town, Ethiopia. Faecal samples and carcasses swabs were collected from cattle systematically selected from the annual plan of Jimma Municipal Abattoir. Personnel hand and knife swabs were collected after slaughtering each selected cattle. A cross-sectional study with systematic sampling method was conducted from October 2019 to September 2020 for the isolation, identification and antimicrobial susceptibility pattern of thermophilic Campylobacter species. Isolation and identification of Campylobacter species were performed according to the techniques recommended by the International Organization for Standardization, and in vitro antibiotic susceptibility testing was screened using the standard agar disc diffusion method as recommended by Clinical and Laboratory Standards Institutions. A total of 684 samples (171 samples from faeces, carcasses, knives and personnel hands, were collected independently). The overall prevalence of thermophilic Campylobacterspecies was 5.6% (38/684). Majority of the isolates were from faecal samples (12.9%, n = 22) followed by carcass swabs(4.1% n = 7), knife swabs(3.5% n = 6) and personnel hand swabs(1.8% n = 3). Isolated and identified species of C.jejuni, C. coli and C. lari accounted for 63.2%, 23.7% and 13.2%, respectively. The isolated Campylobacter species were found to be resistant to Cephalothin (100%), Ampicillin (60.5%), Cefotaxime (60.5%), Chloramphenicol (47.4%) and Tetracycline (42.1%). On the other hand, the isolates were susceptible to Nalidixic acid (86.8%), Ciprofloxacin (86.8%), Sulphamethazole (84.2%), Ceftriaxone (78.9%), Clindamycin (68.4%) and Cefixime (65.8%). 84.2% of the isolates showed multi-drug resistance for three-to-six drug classes. All the C. lari isolates were multidrug resistant. All the three isolated species of Campylobacter were resistant to Cephalothin, and most were multidrug resistant. Isolation of Campylobacter species from faecal, carcass, knife and hand swabs revealed possible risk of contamination and exposure to Campylobacter infection of those who consume raw meat. Therefore, enactment of hygienic practices during the slaughtering process, proper handling and cooking of meat and awareness creation on jurisdictional antibiotic usage are required to avoid Campylobacter infection.

Thermophilic Campylobacter species are common cause of animal and human bacterial diseases with growing resistance to antimicrobials. The aim of this study was to determine the prevalence and antimicrobial susceptibility pattern of Campylobacter species from bovine, knives and personnel in Jimma Town, Ethiopia. Faecal samples and carcasses swabs were collected from cattle systematically selected from the annual plan of Jimma Municipal Abattoir. Personnel hand and knife swabs were collected after slaughtering each selected cattle. A cross-sectional study with systematic sampling method was conducted from October 2019 to September 2020 for the isolation, identification and antimicrobial susceptibility pattern of thermophilic Campylobacter species. Isolation and identification of Campylobacter species were performed according to the techniques recommended by the International Organization for Standardization, and in vitro antibiotic susceptibility testing was screened using the standard agar disc diffusion method as recommended by Clinical and Laboratory Standards Institutions. A total of 684 samples (171 samples from faeces, carcasses, knives and personnel hands, were collected independently). The overall prevalence of thermophilic Campylobacterspecies was 5.6% (38/684). Majority of the isolates were from faecal samples (12.9%, n = 22) followed by carcass swabs(4.1% n = 7), knife swabs(3.5% n = 6) and personnel hand swabs(1.8% n = 3). Isolated and identified species of C.jejuni, C. coli and C. lari accounted for 63.2%, 23.7% and 13.2%, respectively. The isolated Campylobacter species were found to be resistant to Cephalothin (100%), Ampicillin (60.5%), Cefotaxime (60.5%), Chloramphenicol (47.4%) and Tetracycline (42.1%). On the other hand, the isolates were susceptible to Nalidixic acid (86.8%), Ciprofloxacin (86.8%), Sulphamethazole (84.2%), Ceftriaxone (78.9%), Clindamycin (68.4%) and Cefixime (65.8%). 84.2% of the isolates showed multi-drug resistance for three-to-six drug classes. All the C. lari isolates were multidrug resistant. All the three isolated species of Campylobacter were resistant to Cephalothin, and most were multidrug resistant. Isolation of Campylobacter species from faecal, carcass, knife and hand swabs revealed possible risk of contamination and exposure to Campylobacter infection of those who consume raw meat. Therefore, enactment of hygienic practices during the slaughtering process, proper handling and cooking of meat and awareness creation on jurisdictional antibiotic usage are required to avoid Campylobacter infection.

Introduction. Campylobacter is one of the leading bacterial causes of food-borne disease. The prevalence of Campylobacter species resistant to antimicrobial agents is increasing. This study is intended to determine prevalence and antimicrobial susceptibility patterns of Campylobacter species among under-five children with diarrhea. Methodology. A cross-sectional study was conducted among 227 under-five children with diarrhea from July to October 2012 at Jimma town. Isolation and identification of Campylobacter species were performed using standard bacteriological techniques. Antimicrobial susceptibility test was performed following standard protocol. Chi-square and Fisher's exact tests were used for analysis. Results. From 227 under-five children, 16.7% were positive for Campylobacter spp.; isolates, C. jejuni, C. coli, and C. lari, accounted for 71.1%, 21.1%, and 7.9%, respectively. Higher rate of resistance was observed to ampicillin 76.3%, trimethoprim-sulfamethoxazole (68.4%), tetracycline (39.5%), chloramphenicol (31.6%), clindamycin (26.3%), and doxycycline (23.7%). Erythromycin, ciprofloxacin, gentamicin, norfloxacin, and nalidixic acid were effective for more than 80% of the isolates. Multiple drug resistance was observed among 78.9% of all the three spp. Conclusions. Isolation rate of Campylobacter spp. was high. C. lari was reported for the first time at this study area. Higher rate of resistance was observed to the commonly used drugs.

BackgroundUnder-five children are at an increased risk for foodborne illnesses because of the ingenuousness of their immune system. Although Campylobacter species are one of the bacterial etiologies of gastroenteritis, Campylobacter gastroenteritis among under-five children is not well considered in Ethiopia. Therefore, this study aimed at exploring the prevalence, associated risk factors, and antibiotic susceptibility patterns of Campylobacter species among under-five children with diarrhea.MethodsThe institution-based cross-sectional study was conducted among under-five children with diarrhea at Jimma Medical Center, southwestern Ethiopia from January 5 to April 21, 2020. Stool samples were collected and inoculated into Campylobacter agar medium. Isolation and identification were done using standard bacteriological techniques. Antibiotic susceptibility testing was conducted on Mueller-Hinton agar supplemented with 10% sheep blood using disk diffusion techniques. Bivariate and multivariate logistic regressions were used to assess the associated risk factors.ResultsA total of 214 under-five children were enrolled. The prevalence of Campylobacter infection was 8.9%. Absence of caretakers’ handwashing before preparation of food [AOR = 3.7, 95% CI: (1.2–10.8)], direct contact with domestic animals [AOR = 3.6, 95% CI: (1.0–12.7)], and consumption of raw dairy products [AOR = 4.5, 95% CI: (1.4–13.9)] are the factors associated with Campylobacter infection. Some Campylobacter species were found to be resistant to most available antibiotics.ConclusionThe magnitude of Campylobacter gastroenteritis indicates the need for routine isolation and identification of Campylobacter species from all under-five children clinically diagnosed with diarrhea. Species that are resistant to the drug of choice for Campylobacteriosis are also emerging. Health education on the importance of pasteurization of milk and caretakers’ handwashing can mitigate the transmission. Mechanism of handling of domestic animals should be considered to reduce transmission of zoonotic diseases like Campylobacteriosis.

The present study was undertaken with the aim of investigating the physical parameters (e.g. organoleptic and specific gravity of raw milk) and also to study the microbiological quality of raw milk (total viable count, Coliform count and Staphylococcal count) from different villages and Bangladesh Agricultural University (BAU) Dairy Farm of Mymensingh District of Bangladesh, during the period from July to November 2007. A total number of 100 raw milk samples were collected at morning and evening from BAU dairy farm and surrounding four villages of BAU campus. The organoleptic and bacteriological qualities of each sample were analyzed. The organoleptic examination included taste panel score to assess consumer's acceptance and the bacteriological analysis comprised enumeration of total viable count (TVC), total colifrom count (TCC) and total staphylococcal count (TSC) for the determination of sanitary quality. The organoleptic quality of the milk samples is more or less same except the Churkhai milk samples which had flat taste (in 16% milk sample). The average values of TVC/ml were log 5.920, 5.934, 6.007, 6.075 and 6.127 for BAU Dairy Farm, Boira, Shutiakhali, Churkahai and Paglabazar respectively; coliform count were log 2.501, 2.522, 2.550, 2.620 and 2.619 respectively; staphylococcal count were log 2.832, 2.812, 2.866, 2.931 and 2.988 respectively. So, it may be concluded that the raw milk samples of BAU Dairy Farm were superior to others collected from the selected villages which may be due to maintaining better hygienic condition. Key words: Raw milk, physical and microbial quality  doi: 10.3329/bjvm.v6i2.2339 Bangl. J. Vet. Med. (2008). 6 (2): 217-221

Isolation and identification of Campylobacter species

Isolation and identification of Campylobacter species in apparently healthy Ostriches

Aims: To investigate carriage and contamination rates of chicken broiler meat, the factors that are associated with Campylobacter spp. colonization and its phenotypic and genotypic antimicrobial resistance from Thika small-scale poultry farms.
 Study Design: The study design was cross-sectional and laboratory based, it employed simple random sampling across 18 small-scale farms.
 Site and Duration of Study: The study was conducted between August and December 2017 at Thika sub-county, a town located 42 Km North East of Nairobi.
 Methodology: One hundred and eighty five cloaca swab samples from live broilers and 158 neck swab samples from broiler carcasses were collected. Isolates were obtained by plating method using mCCDA, conventional methods and duplex PCR were used for the isolation and identification of Campylobacter species.
 Results: Carriage prevalence was at 15.67%, significantly (P = .000) lower than contamination prevalence detected at 30.37%. While the overall Campylobacter spp. prevalence was 22.45%. Risk of Campylobacter colonization in the flock doubled in feeding broilers with chicken waste and older poultry, at (OR: 2.57, 95% CI: 0.19 - 34.47) and (OR: 2.00, 95% CI: 0.312 - 12.84) respectively. The Campylobacter spp. were resistant (P < .05) against Ciprofloxacin, Streptomycin, and Trimethoprim between carriage and contamination. MDR was 79.22%; XDR was 12.98% while no PDR recorded.
 Conclusion: Broilers in Thika region are potentially important source of human infection and possible continuity of infection from the threat posed by Campylobacter carrier broilers. Presence of sulI and dhfr genes with high resistance observed for quinolones, sulfonamides, ß-lactams and trimethoprim, thus posing a major public health problem for consumers of poultry products.

Introduction: Campylobacter are zoonotic bacteria that cause gastroenteritis worldwide with the species, Campylobacter jejuni and Campylobacter coli commonly associated with human diarrhea. Transmission is mainly through direct contact with farm animals, consumption of chicken and contaminated water. There is paucity of data on the epidemiology of Campylobacter in developing countries despite its global widespread and expansion of poultry farming; hence there is the need to explore and build on the available data. This study aimed at determining prevalence and homestead risk practices associated with Campylobacter infection in diarrheal patients in Busia County. Methods: A cross-sectional study was conducted from February, 2017 to April, 2019. Stool samples were collected from patients of all ages attending Busia County referral Hospital and structured questionnaires on homestead associated risk practices administered. Isolation and identification of Campylobacter species was performed using standard culture method on Modified Charcoal Cefoperazone Deoxycholate medium and confirmed by mPCR. Factors associated with Campylobacter infection were evaluated using logistic regression analysis. Results: A total of 132 (11.6%) Campylobacter comprising 89.2% C. jejuni and 10.8% C. coli were isolated from 1200 diarrhoegenic patients sampled. Isolation rate was higher in children aged 5 years (9.4%). Multilevel logistic models showed that homestead poultry farming was a significant risk associated with Campylobacter infection in 5 years (OR 10.05; 95% CI: 2.60 - 24.29, P Campylobacter infection while those who lived in urban areas (OR 0.47, 95% CI: 0.20 - 0.82, P = 0.041) were protected. Contact with infected diarrheal person from the same household (OR 4.72, 95% CI: 2.10 - 10.52, P = 0.006) and consumption of raw milk (OR 7.14, 95% CI: 1.96 - 18.24, P = 0.001) posed risk among those aged > 5 years respectively. Conclusion: Campylobacter jejuni is the leading cause of Campylobacter infections in diarrheal patients. Personal hygiene awareness of mothers/caregivers and proper animal husbandry especially where livestock-human interaction is common are important practices which require the County government support. Further studies are required on sex specific age difference, other social economic factors, domestic animals and the role played by the environment in the transmission of Campylobacter infection. These would advance knowledge and understanding on source attribution and transmission dynamics for effective control and management of the infection.

The prevalence of claw affections was investigated in 602 cattle in two organized dairy farms during June 2010 to May 2011. Of these cattle 176 were from Bangladesh Agricultural University Dairy Farm (BAUDF) and 426 were from Central Cattle Breeding Station and Dairy Farm (CCBSDF). These animals were studied under 6 groups: calf, heifer, pregnant cow, lactating cow, dry cow and breeding bull. Out of 426 cattle in CCBSDF, 85 animals were affected with various claw affections, the prevalence being 19.95%. The prevalence in BAUDF was 22.72%. Claw affections causing lameness comprised sole ulcer (3.82%), heel erosion (2.49%), interdigital necrobacillosis (2.15%), sand crack (1.82%), hoof overgrowth (3.32%) and white line disease (2.32%). The prevalence of lameness in BAUDF was 13.06% and that in CCBSDF was 11.50%. Sole ulcer (3.82%) was predominantly associated with lameness in both farms while the lowest prevalence of lameness was encountered with interdigital dermatitis (1.39%). The lowest prevalence of lameness was observed in calves in both BAUDF (0.56%) and CCBSDF (0.93%). Pregnant cows were the most vulnerable to claw diseases and manifested lameness of varying degrees. Hind limbs were more frequently affected with claw affections than fore limbs. Animals of over 5 years were mostly affected with foot diseases. Breeding bulls showed 2.27% prevalence of lameness in BAUDF and 1.87% CCBSDF. Among various affections sole ulcer ranked the highest in both the farms. Sand crack and interdigital necrobacillosis registered the lowest prevalence in BAUDF and interdigital dermatitis was the lowest in CCBSDF. Sole ulcer ranked the highest among all claw affections. Pregnant cow were more vulnerable to lameness. Hind limbs were frequently affected with various lesions than the forelimbs.Res. Agric. Livest. Fish.4(3): 193-199, December 2017

Background & Aims: Today, food-borne diseases are known as one of the most important public health concerns in many countries. Campylobacter is one of the most prevalent food-borne pathogens. Raw chicken meat has been reported as the main source of human campylobacteriosis. The present study was conducted to investigate the prevalence of Campylobacter species among broiler carcasses at industrial slaughterhouses in Hamedan province, west of Iran. Materials and Methods: Totally, 100 samples were collected using sterile swabs from chicken skin at the post-scalding stage. The samples were enriched in Brucella broth containing Campylobacter selective supplement and incubated at 42º C for 48-72 hours under microaerophilic conditions. The molecular detection and identification of Campylobacter species were performed by the polymerase chain reaction (PCR) using cadF and Hip primers for detecting Campylobacter species and Campylobacter jejuni, respectively. Results: The results of this study revealed that 81% and 31% of broiler carcasses were positive for the presence of Campylobacter species and C. jejuni, respectively. Conclusion: Due to the high contamination rate of chicken meat with this pathogen, precise hygienic control of poultry meat and an increase in consumer awareness seem necessary to decrease human campylobacteriosis.

This study aimed to analyze the hemato profile of crossbred lactating cows reared at Bangladesh Agricultural University (BAU) Dairy Farm. Thirty-two (32) healthy crossbred lactating cows of Holstein-Frisian (HF), Jersey (J), Sindhi Sahiwal (SS) and Red Chittagong (RCC) were selected for the study and divided into four (04) groups. The cows were allowed to feed and manage according to dairy farm own scheduled-ration. The collected blood samples were analyzed for hematological and biochemical parameters viz. Hemoglobin (Hb), Packed cell volume (PCV), Total leukocyte count (TLC), and Total erythrocyte count (TEC) and Serum urea, Glucose, Albumin, Calcium, and Phosphorus, respectively by using an automated chemistry analyzer. The results revealed that the hematological and biochemical parameters non-significantly (p>0.05) differed between the crossbred groups. From the experiment, the higher amount of Hb (11 g/dL) and PCV (40%) was found in HF and RCC cows, respectively. The TLC was higher in SS and RCC but the TEC was recorded stable along with a slight increment in RCC. As far as the biochemical parameters are concerned, the blood urea concentration was found higher (28 mg/dL) in HF and RCC lactating cows. The same pattern was observed in the case of albumin concentration. On the contrary, the lowest value of Calcium was found in RCC but the Phosphorus concentration showed a little higher in RCC. It can be concluded that the overall herd health was satisfactory based on the blood profile parameters.
 Bang. J. Anim. Sci. 2019. 48 (2): 139-144

Culture and isolation of Campylobacter species by the Linearcount 3MA system

Thermophilic Campylobacter species are recognized as a major cause of acute bacterial diarrhea in humans, with broiler being the most common source of human infection. This study was aimed to evaluate the accuracy of conventional bacteriological method (culture and biochemical) in relation to the gold standard PCR for the identification Campylobacter species from broiler at age of slaughter. A total of 109 fecal samples were collected from broilers (rearing in farms and during evisceration process at slaughterhouse) in Al-Najaf and Al-Qadisiyah Governorates. All samples were collected using modified Cary Blair C&S media then inoculated as soon as possible onto Campylobacter selective plates and incubated under microaerophilic condition at 42°C for 48 hrs. Each small colony with mucoid, grayish, flat or convex, entire or irregular edges and non-hemolytic was selected and further characterized by Gram staining, catalase, oxidase and indoxyl test, then confirmed with latex agglutination test. Out of 109 samples, 36 (33%) suspected growth were selected and phenotypically identified as Campylobacter bacteria. The genotypic identification using a 16S rRNA gene was carried out on 109 DNA samples, an 816bp PCR product was amplified by 18 isolates, therefore, the incidence of Campylobacter isolates in the present study was 16.5%. The present data stated an increased frequency of Campylobacter isolates from chicken during evisceration process at slaughterhouse (44%) while it was significantly lower (8.3%) among broiler farms. The specificity of phenotypic identification of isolates (in comparison to genotypic) was 80.22% which reflect the superiority of PCR assay in the identification of Campylobacter species among recovered isolates. The study concluded that biochemical tests should be replaced with molecular methods for identification of Campylobacter. Also there is a significant incidence of Campylobacter in broilers in Iraq. Surveillance and reporting programs on food of animal origin should be implemented to protect humans from the risk of infection with this zoonotic pathogen.

Enumeration and identification of Campylobacter species in the liver and bile of slaughtered cattle