Prevalence of multidrug-resistant Campylobacter species in wastewater effluents: A menace of environmental and public health concern.

The prevalence of multidrug-resistant Campylobacter species in wastewater effluents presents a formidable challenge at the intersection of environmental sustainability and public health. This study examined the presence of multidrug-resistant Campylobacter in wastewater effluents in the Eastern Cape Province, South Africa, and its implications for environmental ecosystems and public health. Forty-five samples from household effluent (HHE) and wastewater treatment plant effluent (WWTPE) were collected at different geographical locations within the province between April and September 2022. The counts of the presumptive Campylobacter genus ranged from 5.2 × 103 to 6.03 × 104 CFU/mL for HHE and 4.93 × 103 to 1.04 × 104 CFU/mL for WWTPE. About 42.55% of the samples were positive for Campylobacter species. Five virulence determinants including the cadF and wlaN were detected in all the isolates; however, flgR (19.23%), ciaB, and ceuE (15.38%) were less prevalent. The antibiogram profiles of confirmed Campylobacter isolates revealed high resistance (>55%) against all tested antibiotics ranging from 55.77% (nalidixic acid) to 92.30% (erythromycin), and resistance against the other antibiotics followed the order ciprofloxacin (51.92%), azithromycin (50%), and levofloxacin (48.08%). On the contrary, gentamicin was sensitive against 61.54% of the isolates, followed by imipenem (57.69%) and streptomycin (51.92%). The WWTPE's antibiotic resistance index (ARI) was 0.19, lower than the permitted Krumperman threshold of 0.2; and HHE's ARIs were higher. The isolates' respective multiple antibiotic resistance indexes (MARI) varied between 0.08 and 1.00. Among the phenotypically resistant Campylobacter isolates examined, 21 resistance determinants encoding resistance against ß-lactam, carbapenems, aminoglycosides, phenicol, quinolones, tetracyclines, and macrolides were detected, which explains the phenotypic resistance observed in the study. This study concludes that the wastewaters in the study areas are important reservoirs of multidrug-resistant and potentially pathogenic Campylobacter species, suggesting the need for proper treatment of the wastewaters to eliminate the organisms in the effluents before discharge the final effluent to the receiving watershed.

Combined application of bacteriophages with a competitive exclusion culture and carvacrol with organic acids can reduce Campylobacter in primary broiler production.

For reducing Campylobacter (C.) in the food production chain and thus the risk to the consumer, the combined application of different measures as a multiple-hurdle approach is currently under discussion. This is the first study to investigate possible synergistic activities in vivo, aiming at reducing intestinal C. jejuni counts by administering (i) bacteriophages (phages) in combination with a competitive exclusion (CE) product and (ii) carvacrol combined with organic acids. The combined application of the two selected phages (Fletchervirus phage NCTC 12673 and Firehammervirus phage vB_CcM-LmqsCPL1/1) and the CE product significantly reduced C. jejuni loads by 1.0 log10 in cecal and colonic contents as well as in cloacal swabs at the end of the trial (33 and 34 days post hatch). The proportion of bacterial isolates showing reduced phage susceptibility ranged from 10.9% (isolates from cecal content) to 47.8% (isolates from cloacal swabs 32 days post hatch) for the Fletchervirus phage, while all tested isolates remained susceptible to the Firehammervirus phage. The use of carvacrol combined with an organic acid blend (sorbic acid, benzoic acid, propionic acid, and acetic acid) significantly reduced Campylobacter counts by 1.0 log10 in cloacal swabs on day 30 only.

Human Campylobacter spp. infections in Italy.

PURPOSE: Campylobacter is a frequent cause of enteric infections with common antimicrobial resistance issues. The most recent reports of campylobacteriosis in Italy include data from 2013 to 2016. We aimed to provide national epidemiological and microbiological data on human Campylobacter infections in Italy during the period 2017-2021. METHODS: Data was collected from 19 Hospitals in 13 Italian Regions. Bacterial identification was performed by mass spectrometry. Antibiograms were determined with Etest or Kirby-Bauer (EUCAST criteria). RESULTS: In total, 5419 isolations of Campylobacter spp. were performed. The most common species were C. jejuni (n = 4535, 83.7%), followed by C. coli (n = 732, 13.5%) and C. fetus (n = 34, 0.6%). The mean age of patients was 34.61 years and 57.1% were males. Outpatients accounted for 54% of the cases detected. Campylobacter were isolated from faeces in 97.3% of cases and in 2.7% from blood. C. fetus was mostly isolated from blood (88.2% of cases). We tested for antimicrobial susceptibility 4627 isolates (85.4%). Resistance to ciprofloxacin and tetracyclines was 75.5% and 54.8%, respectively; resistance to erythromycin was 4.8%; clarithromycin 2% and azithromycin 2%. 50% of C. jejuni and C. coli were resistant to ≥ 2 antibiotics. Over the study period, resistance to ciprofloxacin and tetracyclines significantly decreased (p < 0.005), while resistance to macrolides remained stable. CONCLUSION: Campylobacter resistance to fluoroquinolones and tetracyclines in Italy is decreasing but is still high, while macrolides retain good activity.

Bacteriological Survey of Fresh Minced Beef on Sale at Retail Outlets in Scotland in 2019: Three Foodborne Pathogens, Hygiene Process Indicators, and Phenotypic Antimicrobial Resistance.

ABSTRACT: The health and economic burden of foodborne illness is high, with approximately 2.4 million cases occurring annually in the United Kingdom. A survey to understand the baseline microbial quality and prevalence of food-related hazards of fresh beef mince on retail sale could inform risk assessment, management, and communication to ensure the safety of this commodity. In such a survey, a two-stage sampling design was used to reflect variations in population density and the market share of five categories of retail outlets in Scotland. From January to December 2019, 1,009 fresh minced beef samples were collected from 15 geographic areas. The microbial quality of each sample was assessed using aerobic colony count and Escherichia coli count. Samples were cultured for Campylobacter and Salmonella, and PCR was used to detect target genes (stx1 all variants, stx2 a to g, and rfbO157) for Shiga toxin-producing E. coli (STEC). The presence of viable E. coli O157 and STEC in samples with a positive PCR signal was confirmed via culture and isolation. Phenotypic antimicrobial sensitivity patterns of cultured pathogens and 100 E. coli isolates were determined, mostly via disk diffusion. The median aerobic colony count and E. coli counts were 6.4 × 105 (interquartile range, 6.9 × 104 to 9.6 × 106) and <10 CFU/g (interquartile range, <10 to 10) of minced beef, respectively. The prevalence was 0.1% (95% confidence interval [CI], 0 to 0.7%) for Campylobacter, 0.3% (95% CI, 0 to 1%) for Salmonella, 22% (95% CI, 20 to 25%) for PCR-positive STEC, and 4% (95% CI, 2 to 5%) for culture-positive STEC. The evidence for phenotypic antimicrobial resistance detected did not give cause for concern, mainly occurring in a few E. coli isolates as single nonsusceptibilities to first-line active substances. The low prevalence of pathogens and phenotypic antimicrobial resistance is encouraging, but ongoing consumer food safety education is necessary to mitigate the residual public health risk.

Prevalence and Antimicrobial Resistance of Campylobacter Species in Diarrheal Patients in Mymensingh, Bangladesh.

Campylobacter enteritis is the leading cause of gastroenteritis in humans worldwide including Bangladesh. The objectives of this study were to estimate the prevalence and antimicrobial-resistance status of Campylobacter spp. in human diarrheal samples collected from Surya Kanta Hospital, Mymensingh, Bangladesh. In this study, we evaluated a total of 330 clinical samples for the presence Campylobacter spp. via cultural and biochemical tests and molecular assays. Furthermore, antimicrobial susceptibility testing for Campylobacter species was accomplished by the standard agar disc diffusion technique against eight commercially available antimicrobial agents. A pretested semistructured questionnaire was used to capture the data on socioanthropological factors from the diarrheal patients. Pearson's chi-square test was performed, and a p value of <0.05 was considered for the level of significance. Nearly one in three diarrheal patients admitted in this hospital were infected with Campylobacter spp. Overall prevalence of Campylobacter spp. was estimated to be 31.5% (104/330) that comprised the prevalence of C. jejuni, 21.8% (n = 72), and C. coli, 9.6% (n = 32). Among the positive cases, the prevalence of Campylobacter was higher in the age group 0-5 years (52%) followed by 6-18 years (42.7%), 19-40 years (34.0%), 41-60 years (25.4%), and >60 years (10.5%). Age, family level's personal hygiene, and involvement with animal husbandry were captured as potential determinants to be associated with the Campylobacter positive status. Among the isolates, 27.3% (n = 20) of C. jejuni and 31.2% (n = 10) of C. coli demonstrated as multidrug-resistant (MDR) to three or more antimicrobial agents. The present study shows that Campylobacter spp. is most prevalent among the hospital-admitted diarrheal patients, and proper measures should be taken to reduce the burden focusing on the potential determinants.

Genomic Characterization of Fluoroquinolone-Resistant Thermophilic Campylobacter Strains Isolated from Layer Chicken Feces in Gangneung, South Korea by Whole-Genome Sequencing.

Thermophilic Campylobacter species of poultry origin have been associated with up to 80% of human campylobacteriosis cases. Layer chickens have received less attention as possible reservoirs of Campylobacter species. Initially, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of two archived Campylobacter isolates (Campylobacter jejuni strain 200605 and Campylobacter coli strain 200606) from layer chickens to five antimicrobials (ciprofloxacin, nalidixic acid, erythromycin, tetracycline, and gentamicin) were determined using broth microdilution while the presence of selected antimicrobial resistance genes was performed by polymerase chain reaction (PCR) using specific primers. Whole-genome sequencing (WGS) was performed by the Illumina HiSeq X platform. The analysis involved antimicrobial resistance genes, virulome, multilocus sequence typing (MLST), and phylogeny. Both isolates were phenotypically resistant to ciprofloxacin (MIC: 32 vs. 32 µg/mL), nalidixic acid (MIC: 128 vs. 64 µg/mL), and tetracycline (MIC: 64 vs. 64 µg/mL), but sensitive to erythromycin (MIC: 1 vs. 2 µg/mL) and gentamicin (MIC: 0.25 vs. 1 µg/mL) for C. jejuni strain 200605 and C. coli strain 200606, respectively. WGS confirmed C257T mutation in the gyrA gene and the presence of cmeABC complex conferring resistance to FQs in both strains. Both strains also exhibited tet(O) genes associated with tetracycline resistance. Various virulence genes associated with motility, chemotaxis, and capsule formation were found in both isolates. However, the analysis of virulence genes showed that C. jejuni strain 200605 is more virulent than C. coli strain 200606. The MLST showed that C. jejuni strain 200605 belongs to sequence type ST-5229 while C. coli strain 200606 belongs to ST-5935, and both STs are less common. The phylogenetic analysis clustered C. jejuni strain 200605 along with other strains reported in Korea (CP028933 from chicken and CP014344 from human) while C. coli strain 200606 formed a separate cluster with C. coli (CP007181) from turkey. The WGS confirmed FQ-resistance in both strains and showed potential virulence of both strains. Further studies are recommended to understand the reasons behind the regional distribution (Korea, China, and Vietnam) of such rare STs.

Prevalence and antimicrobial profiling of Campylobacter spp. isolated from meats, animal, and human feces in Northern of Morocco.

The present work was carried out to understand the occurrence and antimicrobial susceptibility of Campylobacter spp., in various samples in Northern of Morocco. For this purpose, a random sampling was undertaken from butcher shops, traditional markets, and slaughterhouse. First, the research of Campylobacter was performed according to the Moroccan standard NM ISO: 10272-1 (2008). Second, the isolates were identified by biochemical tests and real time PCR. After the biochemical and molecular identification of suspected colonies, a disk diffusion method was executed to determine the sensitivity of Campylobacter spp. against 18 antibiotics. The results showed a moderate prevalence of Campylobacter species (130/466) recovered mainly on the Campylobacter blood base agar, where C. coli (108/130) were more prevalent comparable to C. jejuni (22/130) in poultry and cattle meat, raw milk, cloacal and surface swabs, and stool of patient suffering from diarrhea. The findings supported also the sensitivity of multiplex qPCR to detect Campylobacter strains compared to Moroccan standard NM ISO: 10272-1 (2008). Among our isolates, C. jejuni were the most susceptible strain toward colistin, florfenicol, gentamicin, streptomycin, and erythromycin. Nonetheless, the presence of multidrug Campylobacter resistant strains was highly observed in C. jejuni isolated, particularly, from broiler chickens toward the antibiotic classes of cephalosporin, penicillin, monobactam, quinolone, fluoroquinolone, sulfamide, as well as tetracycline. This may be due to common use of these drugs in veterinary medicine and farms as growth factor, which limits the usefulness of these molecules. Hence, the study highlights the importance of resistance profile monitoring of these pathogens in Northern of Morocco, in order to develop appropriate control measures and to reduce the emergence of multidrug-resistant strains.

Bovine and Pig Carcasses as a Source of Campylobacter in Poland: A Reservoir for Antimicrobial-Resistant Campylobacter coli.

Campylobacter is one of the most common causes of foodborne bacterial infections worldwide. Why poultry has been shown to be one of the most significant sources of these bacteria, ruminants, especially cattle, are also responsible for a high number of human Campylobacter jejuni, and to a lesser extent Campylobacter coli, infections. In this study, bovine and pig carcasses in Poland were investigated for the presence of Campylobacter and for their antimicrobial resistance. A total of 204 swabs from bovine carcasses and 355 swab samples from pig carcasses were tested during 2014-2018. Campylobacter was identified in 129 (36.3%) of the pig and in 11 (5.4%) of the bovine carcasses, respectively. The pig isolates were classified as C. coli (121; 34.1%) or C. jejuni (8; 2.3%), whereas the bovine Campylobacter were identified either as C. jejuni (8; 3.9% isolates) or C. coli (3; 1.5% strains). Resistance of the isolates (n = 140) to erythromycin, ciprofloxacin, nalidixic acid, streptomycin, and tetracycline revealed that the vast majority of C. coli was resistant to streptomycin (106 isolates; 85.5%), tetracycline (97; 78.2%), nalidixic acid (90; 72.6%), and ciprofloxacin (88; 71.0%). Among C. jejuni isolates (n = 16) the resistance rates to all antibiotics were lower than in C. coli, irrespective of the origin. A total of 74 of 121 (61.2%) C. coli isolates from the pig carcasses and one of three such isolates from the bovine samples were multiresistant. Most of the C. coli (64 isolates; 85.3%) had the ciprofloxacin+nalidixic acid+streptomycin+tetracycline resistance profile. The results suggest that pig and bovine carcasses may be an underestimated reservoir of Campylobacter, especially for C. coli in pigs. The high antimicrobial resistance rates of such strains to streptomycin, quinolones, and tetracyclines highlight the need for monitoring of these bacteria in such food and food products.

Establishment of sentinel surveillance of human clinical campylobacteriosis in Ireland.

The aim of this work was the establishment of a national laboratory sentinel surveillance service for human clinical Campylobacter in Ireland. This included detailed genomic molecular epidemiology of Campylobacter for 2019. For February-December 2019, 24 clinical microbiology laboratories in Ireland submitted all PCR/culture-positive clinical Campylobacter spp. specimens to Public Health Laboratory (PHL) Dublin one week out of every four. Antimicrobial susceptibility testing (AST) according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria was carried out for Campylobacter spp. isolates for ciprofloxacin, tetracycline and erythromycin. Batch whole genome sequencing (WGS) was carried out on cultures and analysis was performed to determine species, genotype, identify antimicrobial resistance (AMR) and virulence determinants and identify clusters. A total of 75 isolates and 366 PCR-positive stools were received, and 277 isolates recovered (55.7% recovery from stools). Of 257 isolates characterized by WGS, 86.4% (n = 222) were Campylobacter jejuni, 11.7% (n = 30) Campylobacter coli and 1.9% (n = 5) Campylobacter lari. There were 20 clonal complexes with ST-21 clonal complex most prevalent at 26.8% (n = 69). 50.5% (n = 140) of isolates were susceptible to all three antimicrobials tested. 39.3% (n = 109) isolates were ciprofloxacin resistant, 26.3% (n = 73) tetracycline resistant and two isolates erythromycin resistant. Congruence between phenotypic and genotypic AST was observed. There was 95.9% and 95.6% sensitivity and specificity for WGS to predict ciprofloxacin sensitivity and 98.6% and 99.5% sensitivity and specificity for WGS to predict tetracycline sensitivity. Virulence factors flaA, racR, ciaB and cdtB were detected in all isolates. WGS identified 31 potential clusters for public health alert. This sentinel surveillance of human campylobacteriosis in Ireland establishes the basis for a national reference service. Linking with other partners in a 'One Health' framework will help us better understand sources of infection to reduce disease burden and the threat of AMR.

Antimicrobial resistance and recovery of Salmonella, Campylobacter, and Escherichia coli from chicken egg layer flocks in Canadian sentinel surveillance sites using 2 types of sample matrices.

Eggs are important to the diet of Canadians. This product is one of the supply-managed commodities in Canada, but unlike other commodities, where food safety risks are extensively explored and reported, information on the prevalence of enteric organisms (e.g., Salmonella, Campylobacter) and antimicrobial resistance (AMR) in layers in Canada are limited. This study was conducted to determine the prevalence of select bacteria and the associated AMR patterns in layer flocks using 2 sample matrices. Farms were located within FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance sentinel sites (SS). Fecal samples (Ontario: ONSS1a, ONSS1b) and environmental sponge swabs (British Columbia: BCSS2a) were collected. Salmonella prevalence was 29% and 8% in ONSS1a and ONSS1b, respectively, and 7% in BCSS2a. S. Kentucky and S. Livingstone were the most frequently isolated serovars and no S. Enteritidis was detected. Campylobacter was not detected in the BC sponge swabs but was isolated from 89% and 53% of Ontario fecal samples (ONSS1a and ONSS1b, respectively). Seven C. jejuni from Ontario were ciprofloxacin-resistant. Escherichia coli prevalence was high in both sample types (98%). Overall, tetracycline resistance among E. coli ranged from 26% to 69%. Resistance to ceftiofur (n = 2 isolates) and gentamicin (n = 2) was relatively low. There were diverse resistance patterns (excludes susceptible isolates) observed among E. coli in Ontario (10 patterns) and British Columbia (14 patterns). This study revealed that fecal samples are more informative for farm-level monitoring of pathogen and AMR prevalence. Without further validation, sponge swabs are limited in their utility for Campylobacter detection and thus, for public health surveillance.

Les oeufs sont importants pour l'alimentation des Canadiens. Ce produit est l'un des produits soumis à la gestion de l'offre au Canada, mais contrairement à d'autres produits, où les risques pour la salubrité des aliments sont largement étudiés et signalés, des informations sur la prévalence des organismes entériques (p. ex. Salmonella, Campylobacter) et la résistance aux antimicrobiens (RAM) chez les pondeuses au Canada sont limitées. Cette étude a été menée pour déterminer la prévalence de certaines bactéries et les patrons de résistance aux antimicrobiens associés dans les troupeaux de pondeuses en utilisant deux matrices d'échantillons. Les fermes étaient situées au sein de FoodNet Canada et des sites sentinelles (SS) du Programme intégré canadien de surveillance de la résistance aux antimicrobiens. Des échantillons de matières fécales (Ontario : ONSS1a, ONSS1b) et des éponges environnementales (Colombie-Britannique : BCSS2a) ont été prélevés. La prévalence de Salmonella était de 29 % et 8 % pour ONSS1a et ONSS1b, respectivement, et de 7 % pour BCSS2a. Salmonella Kentucky et S. Livingstone étaient les sérotypes les plus fréquemment isolés et aucun S. Enteritidis n'a été détecté. Campylobacter n'a pas été détecté dans les éponges de la Colombie-Britannique, mais a été isolé de 89 % et 53 % des échantillons de matières fécales de l'Ontario (ONSS1a et ONSS1b, respectivement). Sept C. jejuni de l'Ontario étaient résistants à la ciprofloxacine. La prévalence d'Escherichia coli était élevée dans les deux types d'échantillons (98 %). Dans l'ensemble, la résistance à la tétracycline chez E. coli variait de 26 % à 69 %. La résistance au ceftiofur (n = 2 isolats) et à la gentamicine (n = 2) était relativement faible. Divers profils de résistance (à l'exclusion des isolats sensibles) ont été observés chez E. coli en Ontario (10 profils) et en Colombie-Britannique (14 profils). Cette étude a révélé que les échantillons fécaux sont plus informatifs pour la surveillance au niveau de la ferme de la prévalence des agents pathogènes et de la résistance aux antimicrobiens. Sans validation supplémentaire, les éponge sont limitées dans leur utilité pour la détection de Campylobacter et donc pour la surveillance en santé publique.(Traduit par Docteur Serge Messier).

A systematic review and meta-analysis reveal that Campylobacter spp. and antibiotic resistance are widespread in humans in sub-Saharan Africa.

INTRODUCTION: Campylobacter spp. are zoonotic bacteria that cause gastroenteritis in humans worldwide, whose main symptom is diarrhea. In certain cases, extra intestinal manifestations may occur, such as Guillain Barré syndrome. The bacteria cause severe diarrhea mostly in children and in immunocompromised individuals. This review aims to address the prevalence of Campylobacter spp. in humans in sub-Saharan Africa. It also aims to understand the impact of HIV in the prevalence, as well as to report data on antibiotic resistance and propose research priorities. METHODS: We followed PRISMA guidelines to find studies on the occurrence of Campylobacter spp. in humans in all countries from sub-Saharan Africa. Studies published between 2000 and 2020 were searched in PubMed, Cochrane Library, CINAHL, African Index Medicus, African Journals Online, Google Scholar and Science Direct. We have conducted a random-effect meta-analysis and calculated the proportion of resistant isolates to different antibiotics. RESULTS AND DISCUSSION: We found 77 studies that described such occurrence in humans in 20 out of 53 sub-Saharan African countries. Campylobacter jejuni was the most prevalent species. Pooled prevalence was 9.9% (CI: 8.4%-11.6%). No major variations within the different sub-regions were found. Most studies reported Campylobacter spp. as the cause of diarrhea, mainly in children. Some studies reported the bacteria as a possible etiologic agent of acute flaccid paralysis and urinary tract infection. Campylobacter spp. presented a higher pooled prevalence in HIV infected patients, although not statistically significant. High proportions of resistant strains were reported for many antibiotics, including erythromycin and tetracycline. CONCLUSION: Campylobacter spp. occur in sub-Saharan Africa, although information is scarce or inexistent for many countries. Research priorities should include investigation of the understudied species; extra intestinal manifestations; the impact of HIV infection and associated risk factors. Control strategies should be reinforced to contain the spread of this pathogen and drug resistance.

Occurrence of Campylobacter spp. in Swedish calves, common sequence types and antibiotic resistance patterns.

AIMS: Cattle are the second most important cause of human campylobacteriosis, after poultry, but there are knowledge gaps regarding Campylobacter in cattle. This study examined the occurrence of Campylobacter, the species present, sequence types and antibiotic resistance in Swedish cattle. METHODS AND RESULTS: Faeces samples collected from 154 calves on seven Swedish farms, and 69 follow-up samples from a second collection occasion, were analysed. Campylobacter were isolated from 77% of calves at the first sampling, with Campylobacter jejuni as the most frequently isolated species. Animals kept on deep straw bedding were less likely to be colonized with Campylobacter. Whole-genome sequencing of 90 C. jejuni samples resulted in 11 sequence types, among which ST-19 and ST-21 were most frequent. Antimicrobial resistance analyses showed that 46% of 142 isolates analysed were resistant to quinolones, while all isolates belonging to ST-19, ST-22 and ST-441 were resistant to ciprofloxacin and nalidixic acid. CONCLUSIONS: Campylobacter jejuni was the species most frequently isolated in calves and a strong association was found between sequence type and antimicrobial resistance pattern. SIGNIFICANCE AND IMPACT OF THE STUDY: The high proportion of calves with quinolone-resistant Campylobacter jejuni should be considered in a One Health perspective.

Effect of antimicrobial use and production system on Campylobacter spp., Staphylococcus spp. and Salmonella spp. resistance in Spanish swine: A cross-sectional study.

Antimicrobial resistance is a worldwide public health threat; hence, current trends tend to reduce antimicrobial use in food-producing animals and to monitor resistance in primary production. This study aimed at evaluating the impact of antimicrobial use and production system on swine farms in the antimicrobial resistance of Campylobacter, Salmonella and Staphylococcus, the main zoonotic pathogens in pig herds, in order to assess their potential value as sentinel microorganisms in antimicrobial resistance surveillance schemes. A total of 37 Spanish swine farms, 18 intensive and 19 organic/extensive farms, were included in the study. The antimicrobial resistance of 104 Campylobacter, 84 Staphylococcus and 17 Salmonella isolates was evaluated using Sensititre plates following the EUCAST guidelines. Mixed-effects logistic regression was used to evaluate the influence of production system and antimicrobial use in resistant and multidrug-resistant (MDR) phenotypes to the antimicrobials tested. The results showed that antimicrobial use was higher (p < .001) on intensive farms than on organic/extensive farms. MDR in Campylobacter and Staphylococcus was lower on organic/extensive farms (OR < .01p < .001). Antimicrobial resistance in Campylobacter and Staphylococcus isolates was, also for most of the antimicrobials studied, significantly higher in intensive than organic/extensive pig herds. Tetracycline resistance was associated with total antimicrobial consumption in both microbial species (p < .05), and some cross-associations between distinct antimicrobial substances were established, for instance resistance to erythromycin was associated with macrolide and phenicol consumption. No significant associations could be established for Salmonella isolates. The results demonstrate the link between antimicrobial consumption and resistance in zoonotic bacteria and evidence the potential value of using Campylobacter and Staphylococcus species in monitoring activities aimed at determining the impact of antimicrobials use/reduction on the occurrence and spread of antimicrobial resistance.

Impact of pH on efficacy of peroxy acetic acid against Salmonella, Campylobacter, and Escherichia coli on chicken wings.

Peroxy acetic acid (PAA) is widely used as an antimicrobial in poultry processing, specifically in the chiller. While the natural pH of PAA at the concentrations used is between 4.5 and 6.0, poultry processors adjust the pH to ≥8.0 to maintain product yield. The objective of this study was to evaluate 1) efficacy of PAA at different concentrations, pH, and contact times against Salmonella, Campylobacter, and Escherichia coli and 2) use of E. coli as a surrogate for Salmonella and Campylobacter to conduct validations studies for poultry processing. Fresh chicken wings (0.45 Kg) were inoculated with a cocktail of nalidixic acid-resistant Salmonella Typhimurium, rifampicin-resistant E. coli (5-strain cocktail), and gentamicin-resistant Campylobacter coli. Inoculated chicken wings were immersed in PAA solutions of 50, 250, and 500 ppm adjusted to pH 8.2 and 10.0 as well as nonadjusted PAA solutions for 10 s and 60 min. Treated chicken wings were rinsed in chilled buffered peptone water, serially diluted, and plated on Petrifilm APC for enumerating Salmonella and E. coli populations and spread plated on Campy Cefex Agar containing gentamicin (200 ppm) to enumerate Campylobacter. Immersion of chicken wings in 500 ppm of PAA (non-pH-adjusted) for 60 min resulted in greater microbial reductions (P ≤ 0.05) of Salmonella, Campylobacter, and E. coli populations of 2.56, 1.90, and 2.53 log CFU/mL, respectively. Higher concentrations and longer exposure times resulted in greater reductions (P ≤ 0.05) of Salmonella, E. coli, and Campylobacter populations, and increasing pH of PAA solution did not affect (P > 0.05) its efficacy. A high correlation (r = 0.93) was observed between E. coli (surrogate) and Salmonella populations suggesting that E. coli can be used as a surrogate for Salmonella for conducting validation studies for antimicrobial efficacy testing in poultry processing.

Efficacy of antimicrobial interventions in reducing Salmonella enterica, Shiga toxin-producing Escherichia coli, Campylobacter, and Escherichia coli biotype I surrogates on non-chilled and chilled, skin-on and skinless pork.

Effect of various antimicrobial interventions on pork carcass cuts - skin-on and skinless, non-chilled and chilled - was studied. Carcass pieces were inoculated with Salmonella enterica, Shiga toxin-producing Escherichia coli (STEC), Escherichia coli pathogen surrogates or Campylobacter spp. Inoculated pieces were assigned to one of the following antimicrobial treatments: 2.5% and 5.0% room temperature lactic acid, 2.5% and 5.0% warm (55 °C) lactic acid, 400 ppm (0.4 mg/mL) room temperature peroxyacetic acid (PAA) or warm (55 °C) water. Treated pieces were sampled before antimicrobial treatment of non-chilled pork tissue, then at 30 m and 24 h post-treatment. For chilled pork, samples were collected after 24 h chilling and 30 m post-treatment. Lactic acid and PAA treatments reduced (P < 0.05) pathogenic and surrogate bacteria; warm water did not produce similar results. Objective and sensory color evaluations on treated pork indicated minimal negative impacts on pork color. Various antimicrobial interventions were effective in reducing surrogates on pork without diminishing quality.

Resistance patterns to C and D antibiotic categories for veterinary use of Campylobacter spp., Escherichia coli and Enterococcus spp. commensal isolates from laying hen farms in Spain during 2018.

Antimicrobial Resistance (AMR) is a global threat for human and animal health. Few studies have been carried out on laying hens. The aim of this work was to evaluate the antimicrobial susceptibility of commensal Campylobacter spp., E. coli, and Enterococcus spp. isolates in Spanish laying hens in 2018. Samples were collected from 39 laying hen farms. The microorganisms of interest were isolated and confirmed by PCR. The Minimum Inhibitory Concentration (MIC) to antimicrobials of C and D categories were determined. 195 E. coli, 195 Enterococcus spp. and 25 Campylobacter spp. isolates were obtained. E. coli isolates showed high resistance to D category antimicrobials (sulfamethoxazole 76.41 %, tetracycline 62.05 %, trimethoprim 50.77 %, ampicillin 30.77 %) and lower resistance to C category (azithromycin 30.26 %, gentamicin 12.31 %, chloramphenicol 4.62 %). A 10.26 % of E. coli isolates were susceptible to all antimicrobials tested, Multi Drug Resistance (MDR) to 3 antimicrobial families was found in 23.08 % of the isolates and 13.85 % were MDR to 4 families, being Erythromycin-Sulfamethoxazole-Tetracycline the most common resistance profile (10.77 %). Enterococcus spp. showed very high resistance to D category tetracycline (78.47 %) and C category erythromycin (76.42 %). The 11.79 % of Enterococcus spp. isolates were susceptible to all antimicrobials and 53.33 % were resistant to 2 families, being Erythromycin-Tetracycline the most common AMR profile (51.79 %). Regarding Campylobacter spp., resistance to tetracycline (48 %) was higher than resistance to C category antimicrobials (erythromycin 28 %, streptomycin 24 %, gentamicin 16 %). There was a 52 % sensitivity to all tested antimicrobials and 24 % showed MDR to aminoglycosides, macrolides and tetracyclines (Gentamicin-Streptomycin-Erythromycin-Tetracycline MDR profile). Novel data on AMR in laying hen commensal isolates in Spain was provided. High resistance to several antimicrobials was found, especially to key drugs for the treatment of zoonosis, which represents a public health risk. Better surveillance and careful regulation of antimicrobial use is required in laying hen production.

Prevalence and antimicrobial susceptibility of Campylobacter isolated from retail chickens in Thailand.

Campylobacter is an important foodborne pathogen causing bacterial gastroenteritis worldwide; however, there has been a lack of information over the past decade on its occurrence, antibiotic susceptibility and genetic diversity in Thailand. Poultry meat is considered as a reservoir for transmission of Campylobacter to humans. This study determines the prevalence and antimicrobial resistance patterns of Campylobacter spp. on chicken samples purchased from 50 local wet markets and supermarkets in central Thailand. Of the 296 samples, 99 (33.5%) were contaminated with C. jejuni, 54 (18.2%) were C. coli and 15 (5.1%) were contaminated with both species. Antibiotic resistance rate is higher among C. coli isolates; 100%, 76.8%, 37.7%, 36.2% and 13.0% were resistant to quinolones, cyclines, macrolides, clindamycin and gentamicin, respectively. Most of the C. jejuni isolates were resistant to quinolones (79.8%) and cyclines (38.6%) whereas resistance to macrolides, clindamycin and gentamicin was found to be 1.8%. Multi-drug resistance (i.e. to three or more unrelated antimicrobials) was detected in 37.7% of C. coli and 1.8% of C. jejuni isolates. This study has revealed high contamination rates and alarming levels of antimicrobial resistance in Campylobacter spp. isolated from retail chicken samples in Thailand, suggesting the necessity of implementing interventions to reduce its prevalence from farm to table in the country.

Prevalence of Campylobacter species in human, animal and food of animal origin and their antimicrobial susceptibility in Ethiopia: a systematic review and meta-analysis.

BACKGROUND: Campylobacter jejuni and Campylobacter coli accounts for most cases of human gastrointestinal infections. The infection occurs through ingestion of contaminated food or water, and direct contact with feces of infected animal or human. Regardless of few local reports of Campylobacter and its antimicrobial susceptibility profile, there is no comprehensive data that show the burden of Campylobacter infection at national level in Ethiopia. This systemic review and meta-analysis aimed to determine the pooled prevalence of Campylobacter and its resistance patterns in Ethiopia from different sources. METHOD: A comprehensive literature search of PubMed, Google scholar, Science direct and Google engine search was conducted for studies published from 2000 to July 30, 2020 on prevalence and antimicrobial susceptibility of Campylobacter in human, animal and food. The study was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) Checklist. Data from articles was extracted using a standardized data extraction format. The quality of the studies was assessed based on the Newcastle-Ottawa scale. The Q test and I2 test statistic were used to test heterogeneity across studies. The Pooled estimate of prevalence of Campylobacter species and its antimicrobial susceptibility profile was computed by a random effects model using STATA 16.0 software. Results were presented in forest plot, tables, funnel plot and figures with 95% confidence interval. RESULTS: A total of 291 articles were retrieved initially. The pooled prevalence of Campylobacter species from different sources was 10.2% (95% CI 3.79, 16.51). In this meta-analysis, the lowest prevalence was 6.0% whereas the highest prevalence was 72.7%. In the sub-group analysis, the pooled prevalence was similar in Amhara and Oromia region, higher in Gambella and lower in Sidama. Prevalence of Campylobacter was higher in animals (14.6%) compared to humans (9%). The pooled antimicrobial resistance rates of Campylobacter species to different antimicrobials ranged from 2.9-100%. Overall, higher rate of resistance was to cephalothin (67.2%), gentamicin (67.2%), and trimethoprim-sulfamethoxazole (33.3%) in Campylobacter isolates from all sources. In isolates from human, resistance to cephalothin was 83% followed by amoxicillin (80%), amoxicillin-clavulnate (36%), trimethoprim-sulfamethpxazole (32%), clindamycin (31%) and ceftriaxone (28%). On the other hand, higher rate of resistance to penicillin (100%), cephalothin (60%), ciprofloxacin (71.2%), and trimethoprim-sulfamethoxazole (39%) was recorded in isolates from animals. CONCLUSION: The present study highlights the burden of Campylobacter species in the country and higher rate of resistance among investigated isolates. Designing appropriate prevention strategies and further local in-depth studies are recommended to establish actual epidemiological burden of the bacteria in the country.

Characterization of Posttranslationally Modified Multidrug Efflux Pumps Reveals an Unexpected Link between Glycosylation and Antimicrobial Resistance.

The substantial rise in multidrug-resistant bacterial infections is a current global imperative. Cumulative efforts to characterize antimicrobial resistance in bacteria has demonstrated the spread of six families of multidrug efflux pumps, of which resistance-nodulation-cell division (RND) is the major mechanism of multidrug resistance in Gram-negative bacteria. RND is composed of a tripartite protein assembly and confers resistance to a range of unrelated compounds. In the major enteric pathogen Campylobacter jejuni, the three protein components of RND are posttranslationally modified with N-linked glycans. The direct role of N-linked glycans in C. jejuni and other bacteria has long been elusive. Here, we present the first detailed account of the role of N-linked glycans and the link between N-glycosylation and antimicrobial resistance in C. jejuni We demonstrate the multifunctional role of N-linked glycans in enhancing protein thermostability, stabilizing protein complexes and the promotion of protein-protein interaction, thus mediating antimicrobial resistance via enhancing multidrug efflux pump activity. This affirms that glycosylation is critical for multidrug efflux pump assembly. We present a generalized strategy that could be used to investigate general glycosylation system in Campylobacter genus and a potential target to develop antimicrobials against multidrug-resistant pathogens.IMPORTANCE Nearly all bacterial species have at least a single glycosylation system, but the direct effects of these posttranslational protein modifications are unresolved. Glycoproteome-wide analysis of several bacterial pathogens has revealed general glycan modifications of virulence factors and protein assemblies. Using Campylobacter jejuni as a model organism, we have studied the role of general N-linked glycans in the multidrug efflux pump commonly found in Gram-negative bacteria. We show, for the first time, the direct link between N-linked glycans and multidrug efflux pump activity. At the protein level, we demonstrate that N-linked glycans play a role in enhancing protein thermostability and mediating the assembly of the multidrug efflux pump to promote antimicrobial resistance, highlighting the importance of this posttranslational modification in bacterial physiology. Similar roles for glycans are expected to be found in other Gram-negative pathogens that possess general protein glycosylation systems.

Antimicrobial interventions to reduce Salmonella and Campylobacter populations and improve shelf life of quail carcasses.

Quail (Coturnix japonica) is processed and marketed as fresh meat, with limited shelf life. The objective of this study was to evaluate the efficacy of antimicrobial interventions during slaughter on reducing Salmonella and Campylobacter contamination and to determine the microbiological shelf life of quail during refrigerated (4°C) storage. Three antimicrobials, peracetic acid (400 ppm; PAA), Citrilow (pH 1.2), and Cecure (cetylpyridinium chloride [CPC], 450 ppm), along with a water and no-treatment control were evaluated. Quail carcasses (n = 75) were inoculated with a cocktail of nalidixic acid-resistant Salmonella Typhimurium and gentamicin-resistant Campylobacter coli. After 30 min of attachment time, quail carcasses were submerged in each antimicrobial solution for 20 s with air agitation. Noninoculated quail carcasses (n = 25) were similarly treated, packaged, and stored under refrigeration (4°C). Aerobic plate counts (APC), psychrotroph counts (PC), Enterobacteriaceae counts (ENT), total coliform counts (TCC), and Escherichia coli counts on quail carcasses were determined on 1, 4, 7, and 10 d. Salmonella and Campylobacter populations were determined by plating on Petrifilm APC supplemented with 200-ppm nalidixic acid and Campy Cefex agar supplemented with 200-ppm gentamycin, respectively. No significant reductions in (P > 0.01 log cfu/mL) in APC, PC, ENT, TCC, and E. coli counts were observed on carcasses submerged in water. However, treatments with PAA, Citrilow, and CPC significantly reduced (P ≤ 0.05) Salmonella and Campylobacter coli contamination. Citrilow showed greater (P ≤ 0.05) reduction in Salmonella and Campylobacter population (1.90 and 3.82 log cfu/mL reduction, respectively) to PAA and CPC. Greater (P ≤ 0.05) reductions in APC, PC, ENT, TCC, and E. coli counts (2.22, 1.26, 1.47, 1.52, and 1.59 log cfu/mL, respectively) were obtained with the application of CPC. Application of antimicrobial interventions resulted in a reduction in Campylobacter and Salmonella, APC, PC, and ENT populations after treatments (day 0) and throughout the storage period (day 10). Use of antimicrobial interventions after slaughter can improve the microbiological safety and shelf life of quail.