Slaughterhouse Environment Research Articles

Aureimonas altamirensis: First Isolation from a Chicken Slaughterhouse in Italy Followed by Genotype and Phenotype Evaluations

The presence of foodborne pathogens in meat is linked to several contamination sources, and the slaughterhouse environment represents a relevant reservoir of contamination. Aureimonas altamirensis is a Gram-negative bacteria associated with different isolation sources, including human clinical cases. This study aims to identify and characterize an A. altamirensis isolate from chicken guts collected in an Italian slaughterhouse. The study approach includes whole-genome analysis jointly with phenotypical tests. Whole-genome sequencing (WGS) confirms the initial MALDI-TOF MS identification, finding putative virulence and biofilm-related genes. Moreover, the gene class evaluation reveals that the numerically largest gene category in the A. altamirensis genome is related to amino acid metabolism and transport. The analyses performed on a human gut mucus-producing cell line (HT29-MTX-E12) demonstrated the ability of A. altamirensis to colonize the host cell layer. Moreover, the antibiotic resistance test showed a high resistance of A. altamirensis to gentamicin (MIC 0.5 mg/L). The detection of a potential pathogenic and antibiotic-resistant A. altamirensis strain isolated from a slaughterhouse underlines the necessity of active surveillance studies focused on this species and the need for further studies about A. altamirensis in foods.

Environmental impact assessment of abattoirs in Rivers State, Nigeria

Abattoir waste discharged randomly into a recipient ecosystem can have a serious ecological impact. This study aimed to assess the environmental impact of abattoir operations, looking at the physicochemical and microbiological quality of air, soil and wastewater within the vicinity of operations. Physicochemical parameters of air, soil and water were found to differ across locations, though not significant (p>0.05). BOD and COD exceeded the tolerable limits of 30-100 mg/l and 250 mg/l respectively, for all samples. Only the pH (10.62) of wastewater from the Mbodo-Aluu abattoir exceeded the tolerable limit of 5.5-9.0. All heavy metals were within tolerable limits, except Zn, which exceeded 0.2 mg/l. Microbiological parameters of air, soil and water were also found to differ across locations, though not significant (p>0.05). Abattoir operations impact the physicochemical characteristics of wastewater, soil, and air in the abattoir environment. Microbes associated with wastewater, soil impacted by abattoir wastewater, and air from the abattoir environment were found to include pathogenic species of bacteria and fungi. Proper waste management should be encouraged among abattoir operators and should include treatment of wastewater.

Plasmid-Borne Mobile Colistin Resistant Gene (Mcr-1) Detection and Multidrug Resistant Bacteria Isolated from Some Abattoir Environments in Benin City, Edo State, Nigeria

Antibiotics resistance is an increasing public health challenge globally, and very recently, global attention has focused on colistin, which is termed “last resort antibiotics”. This study was aimed at investigating the prevalence of plasmid-borne mobile colistin resistant and multidrug resistant bacteria from 6 major abattoirs located in Benin City, Edo State, Nigeria. Two hundred and eighty-eight samples from fresh water, wastewater, utensils, and handlers were obtained over a 6-months period. Mean mesophilic aerobic bacteria (MAB) and thermotolerant coliform bacteria (TCB) were determined by pour plate method, while the indoor air of the abattoirs was sampled using passive sedimentation technique. Bacterial isolates were identified by morphological, biochemical and 16S rRNA analysis. Phenotypic detection of colistin-resistance as well as multi-drug resistant profile of all isolates was done by the modified Kirky Bauer method. The presence and/or absence of colistin-resistance gene (mcr-1 to mcr-8) were investigated by polymerase chain reaction. The MAB ranged from 0.3 ± 0.0 cfu/m3 in indoor air from both Holy Ghost B and Bob Izua abattoir to 2.6 ± 0.3 cfu/ml in wash water from Holy Ghost A abattoir, while the TCB ranged from 0.0 ± 0.0 cfu/ml in wastewater from Lawal and Sons abattoir to 0.6 ± 0.1 cfu/ml in wash water from Osazee abattoir. A total of 149 bacterial isolates, belonging to 6 different species (Pseudomonas aeruginosa PA01, Enterobacter ludwigii EN-119, Providencia stuartii PRV00010, Klebsiella quasipneumoniae strain KqPF26, Enterococcus saccharolyticus ATCC 43076 and Provincia rettgeri strain AR_0082) were obtained with the majority (>90%) being multidrug resistant. Seven (4.7%) of the isolates were phenotypically resistant to colistin, while only 3 harbored the mcr-1 gene. This result shows that plasmid-borne colistinresistant and multidrug resistant bacteria are prevalent in abattoir environment located in Benin City, Edo State, Nigeria. This is an indication that abattoir facilities could be a source of human exposure to colistin resistant bacteria, and efforts must be made at reducing the high dependence of antibiotics in farm animals.

Biofilm Forming Capacity and Presence of Biofilm-Associated Virulence Genes of Enterococcus faecalis isolates from Slaughterhouse Environments

Bu çalışmada, Kayseri ilinde mevcut olan birinci sınıf bir sığır mezbahasında kesim hattı boyunca alınan örneklerde biyofilm oluşturma yeteneğine sahip E. faecalis varlığı ve izolatlar da biyofilm ile ilişkili virülans faktör genlerinin araştırılması amaçlandı. Çalışmada, kesimhaneden alınan toplam 300 adet örnekte (180 karkas, 102 ekipman ve yüzey ve 18 mezbaha atık suyu) etkenin varlığı konvansiyonel yöntem ve PZR ile analiz edildi. İzolatların biyofilm oluşturma yeteneklerini belirlemede Kongo kırmızısı agar ve mikroplaka testi ve biyofilm ile ilişkili virülans genlerinin (gelE ve esp) tespitinde PZR kullanıldı. Analiz edilen 300 örneğin 40’ından (%13.3) E. faecalis izole edildi ve bu izolatların 35 (%87.5) biyofilm oluşturma yeteneğinde idi. Biyofilm pozitif olan izolatların 33’ünde (%82.5) gelE geni belirlendi, ayrıca bu izolatların bir tanesini (% 2.5) esp genini de içeriyordu. Sonuç olarak, Kayseri ilindeki kesimhaneden alınan tüm örnek türlerinde mezbahanın hijyenik kalitesini yansıtan biyofilm oluşturma yeteneğine sahip E. faecalis’in tespit edilmesi ve izolatların biyofilm oluşumunun virülans faktör genler ile ilişkisi tespit edilmesi gıda güvenliği açısından önemlidir.

Transmission of linezolid-resistant Enterococcus isolates carrying optrA and poxtA genes in slaughterhouses

IntroductionThe presence of linezolid-resistant enterococci found in animal-derived food has attracted attention for possible transmission to human-derived enterococci through the food chain. Linezolid-resistant enterococci in farms have been widely reported, but enterococci carrying antimicrobial resistance (AMR) genes poxtA, optrA, or cfr(D) in slaughterhouse environments have not been well addressed.MethodsEnterococcus was isolated from the samples collected from two slaughterhouses in Hangzhou, and the Enterococcus carrying linezolidin-resistant genes was identified by PCR. The minimum inhibitory concentration (MIC) of the Enterococcus carrying linezolidin-resistant genes was determined by microbroth dilution method. Finally, the whole genome of strains carrying two or more linezoline resistance genes was sequenced using the Oxford Nanopore TechnologyResultsHere, 291 enterococci strains were isolated from 309 samples (94.17%). A total of 4 poxtA-positive enterococci and 42 optrA-positive enterococci were identified based on PCR. The antimicrobial susceptibility test showed that the highest rate of florfenicol resistance was 97.82% and the rate of multidrug resistance (MDR) was 95.65%. Two strains carried multiple linezolid resistance genes, among which Enterococcus casseliflavus CQFYY22-063 cocarrying optrA, poxtA, and cfr(D) was isolated from the duck cecum, and Enterococcus faecium CQFYH22-006 cocarrying optrA and poxtA was isolated from slaughterhouse sewage for the first time. Furthermore, Oxford Nanopore Technology revealed that the optrA gene of strain CQFYY22-063 was located on the Inc18-type plasmid pFYY063-optrA-70K, and the poxtA and cfr(D) genes were located on the Inc18-type plasmid pFYY063- poxtA-12K. Meanwhile, the poxtA gene of strain CQFYH22-006 was located on the Rep3-type plasmid pFYH006-poxtA-25K, and the optrA gene was located on the chromosome.DiscussionTogether, linezolid resistance in slaughterhouses deserves extensive attention, indicating the need to strengthen the monitoring of different links in the food production chain within the One Health concept.

Evaluation of Pollutant Discharge from Slaughterhouses on Receiving Water Bodies

Aim: The aim of this study is to appraisal the consequences of slaughterhouse wastewater discharge into surface water of selected slaughterhouse environment.
 Study Design: Three (3) main stream/surface water discharged slaughterhouses was analyzed. Water samples were collected at the Middle Course (MC) of the receiving water body. Sampling was be carried out between the hours of 10.00 am and 11.00 am when the discharge of effluent into the rivers/streams always occur.
 Methodology: Wastewater samples were collected from the different slaughterhouses in order to determine the quality of the effluent before it was discharged into nearby stream/river. The MC samples will be collected from about 40 m from the point of entry of the slaughterhouse waste. Waste samples were collected from each sampling site with a clean polyethylene bottles. The containers was washed with diluted HCl and then rinsed with the wastewater from the slaughterhouse so as to neutralize or reduce the effect of external contaminants. The samples were appropriately labeled and transported to the laboratory, stored in the refrigerator at 4⁰C prior to being analyzed for the physical, chemical and biological parameters present.
 Results: The parameters assessed include, Biochemical Oxygen Demand (BOD), Chemical Oxygen Demand (COD), Total Solids (TS), Phosphate, pH, Magnesium, Zinc, Iron, Manganese, Cadmium and Lead. High level of BOD, COD, and TS was revealed on receiving streams with minimum values of 1001mg/l, 1120mg/l, and 990mg/l.
 Conclusion: The physico-chemical parameters showed the negative impact of the abattoir effluent on the stream thus rendering the water of worthless value to humans. Furthermore, the abattoir urgently needs an effluent treatment facility to be installed to reduce the health hazard its effluent poses on the abattoir users and users of the streams receiving the effluent.

Analysis of bacterial diversity associated with commercial broiler chicken in Lahore, Pakistan

Chicken (Gallus gallus domesticus) is the most widely domesticated animal worldwide. Food biosafety is a great concern for consumers, producers, and public health organizations around the world. The poultry sector contributes 3.1% to the GDP of Pakistan. Infections caused by bacteria and viruses in poultry pose a serious threat to consumer health and economic burden in Pakistan while accurate figures are still unknown due to several factors, including inadequate surveillance, management, monitoring, and infection control. The chicken samples were procured from Shadman, Moon, Karim Block, Tollinten, Jallo and Wapda Town meat market of Lahore, Pakistan. Overall, 63 bacterial strains were isolated from complex and selective media and further processed for characterization. The 16S rRNA gene sequencing of bacterial strains were exhibiting genetic similarity with Proteus, Shigella, Providencia, Escherichia, Citrobacter, Klebsiella, Myroides, Enterococcus, Serratia, Staphylococcus, Bacillus, Macrococcus and Kluyvera. Bacterial strains were evaluated for antibiotic susceptibility testing, swarming motility, and biochemical tests. Antibiotic susceptibility was tested by using discs of tetracycline, gentamycin, streptomycin and chloramphenicol. Bacterial strains were recorded as resistant to all antibiotics except chloramphenicol. It was confirmed that chicken meat was infested with potential opportunistic human pathogens. To overcome the foodborne illnesses and to control the microbial load, food safety measures should be taken by following the standard protocols. In Pakistan, slaughterhouse environment, their sanitary conditions, storage, and transportation may be responsible in spreading the pathogens to food products.

Meat processing and handling facilities in slaughterhouses of selected areas of Bangladesh

The study was conducted to investigate the present status of meat processing facilities in different slaughterhouse of Bangladesh through field survey. The data was collected through interview schedule selecting 30 respondents (Butcher, Seller and Consumer) in Khulna, Rajshahi and Rangpur Division in Bangladesh. They were asked about knowledge of hygienic meat production, slaughterhouse environment, existing facilities, sanitation and waste management, opinion of the butcher, seller and consumers. It was obtained from the study indicating that approximately 75.63% of the animals were slaughtered at the abattoir. Only 5.8% of butchers (n=30) knew about the production of hygienic meat. Almost 100% of the abattoirs lacked hygiene and sanitation. Only 26.66% slaughtered carcasses were checked by veterinary surgeon where another 16.7% were checked by unskilled meat inspectors from the municipality. Only 23.33% of slaughterhouses were situated in environmentally friendly locations. Consumer perception revealed that only 3.33% of consumers have knowledge of meat hygiene. Meat is a great source of animal protein in our country but hygienic condition and facilities available in slaughterhouse were not satisfactory. Slaughterhouses both in rural or municipal areas were contaminated frequently and therefore meat and meat byproducts coming from such conditions often deteriorated through bacterial infection or contamination might be the cause of food poisoning or diseases in consumers.
 Bangladesh Journal of Animal Science 51 (3): 115-121, 2022

Distribution of psychrophilic microorganisms in a beef slaughterhouse in Japan after cleaning.

The purpose of this study was to investigate the abundance and distribution of psychrophilic microorganisms associated with spoilage in beef slaughterhouse environments after cleaning. The processing lines and equipment used in slaughtering and boning were swabbed, and the microbial count was determined using a TSA and MRS medium and Chromocult® Coliform agar incubated at 15ºC and 37ºC, respectively. As a result, the brisket saw (handle side) and trolley hook were the most heavily contaminated with microorganisms, with each having a microbial adhesion rate of 66.7%. The microbial adhesion rates of the apron and milling cutter (edge side) were 50%, respectively, and those of the foot cutter (edge and handle side), splitting saw (edge side), and knife (handle side) were 33.3%, respectively. Next, four colonies were randomly isolated from the petri dish used for the bacterial count measurement to identify the predominant microbial species of the microorganisms attached to each equipment. As a result of Sanger sequencing analysis, yeasts such as Candida zeylanoides and Rhodotorula sp. and bacteria including Pseudomonas sp. and Rhodococcus sp. were identified from the equipment used in the slaughtering line, and it was assumed that these microorganisms were of environmental origin. In contrast, only Pseudomonas sp. and Candida zeylanoides were isolated from the boning line. Despite the use of cleaning operations, this study identified some equipment was contaminated with microorganisms. Since this equipment frequently comes into direct contact with the carcass, it is critical to thoroughly remove the microorganisms through accurate cleaning to prevent the spread of microbial contamination on the carcasses.

Prevalence of Livestock-Associated MRSA ST398 in a Swine Slaughterhouse in Guangzhou, China.

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is an important zoonotic microorganism that is increasingly causing public health concern worldwide. The objective of this study was to determine the transmission and occurrence of MRSA in a slaughterhouse environment and evaluate its antimicrobial resistance and genetic characterization. In this study, we conducted a comprehensive epidemiological survey of S. aureus by spa typing and whole-genome sequencing (WGS) of samples obtained from the pork production chain, the environment, and community residents. To clarify the evolutionary relationships of MRSA sequence type (ST) 398 in this study and global isolates, 197 published whole-genome sequences data of MRSA ST398 strains were downloaded from the GenBank database and included in the phylogenetic analysis. A total of 585 porcine samples (snout and carcass swabs), 78 human nasal samples, and 136 environmental samples were collected. The MRSA isolates were detected at higher frequencies in samples from swine (15.0%) than carcasses (10.0%), slaughterhouse workers (8.0%), community residents (0%), and environment samples (5.9%). The spa typing results showed that t571 accounted for a higher proportion than other spa types. Closely related isolates from the samples of swine, slaughterhouse workers, carcasses, carrier vehicle, and surrounding fishpond water indicate that MRSA ST398 strains may spread among swine, humans, and the environment. MRSA ST398-t571 isolates were genetically different from global strains, except for two Korean isolates, which showed genetic closeness with it. In addition, a MRSA ST398 isolate recovered from an infected patient in Europe differed by only 31 SNPs from the airborne dust-associated strain isolated in this study, thereby suggesting potential transmission among different countries. Antimicrobial susceptibility testing results demonstrated that 99.0% (96/97) of MRSA and 95.1% (231/243) of methicillin-sensitive S. aureus (MSSA) showed multidrug-resistant (MDR) phenotypes. According to WGS analysis, the poxtA-carrying segment (IS431mec-optrA-IS1216-fexB-IS431mec) was reported in MRSA ST398 isolates for the first time. The coexistence of cfr and optrA in a plasmid was first detected in MRSA ST398. The potential transmission of MRSA among humans, animals, and the environment is a cause for concern. The emergence and transmission of LA-MRSA ST398 with high levels of resistance profiles highlight the urgent need for LA-MRSA surveillance.

Isolation, discrimination, and molecular detection of Listeria species from slaughtered cattle in Namwala District, Zambia

BackgroundThe food industry is increasingly becoming more scrutinized, given the frequency and intensity with which zoonotic diseases are being reported. Pathogen tracking has become more applicable with regards food safety. It is in this regard that the present study was formulated to track Listeria species. in freshly slaughtered cattle carcasses by utilizing standard and molecular biological techniques.MethodsA cross-sectional study design was conducted from March to December 2020 with 200 samples being equally collected in the rainy and dry seasons. A total of 180 and 20 swabs were aseptically collected from carcasses and the environment respectively. Samples were first subjected to pre-enrichment in half-strength Fraser broth followed by enrichment in full strength Fraser broth and subsequent plating on Listeria agar. Listeria growth characteristics were identified up to species level based on their morphological and biochemical characteristics. Further, molecular detection and phylogenetic analysis was conducted. Quantitative proportionate survey data were analyzed using Stata Version 15 software to estimate crude prevalence taking into account complex design at abattoir level. Factors associated with contamination were characterized using logistic regression. Sequences were analyzed using, Genetyyx version 12 and phylogenetic Mega.ResultsOf the 200 samples, 19 were positive for Listeria species identified as L.innocua 14/19 (73.7%) and L. monocytogenes 5/19 (26.3%). All isolates were from freshly slaughtered carcasses, and none from environment. Siginificant differences in contamination levels were observed based on season: rainy season yielded 14 (73.6%) whilst the dry season 5 (26.3%). The L. monocytogenes strains showed a high degree of homogeneity on phylogenetic analysis and clustered based on abattoir. Seasonality was identified as a major determinant influencing contamination based on the final logistic regression model.ConclusionThis study found evidence of L. monocytogenes contamination on traditionally raised beef carcasses across various abattoirs surveyed. The failure to find Listeria contamination on the abattoir environment may to a greater extent intimate cattle carccases as primary sources of contamination. However, a more comprerehnsive study incorporating different geographical regions is needed to conclusively ascertain these present findings.

Research Note: Persistent Salmonella problems in slaughterhouses related to clones linked to poultry companies

Salmonellosis remains one of the main foodborne zoonoses in Europe, with poultry products as the main source of human infections. The slaughterhouse has been identified as a potential source for Salmonella contamination of poultry meat. Despite the mandatory programme of the EU, there are companies with persistent Salmonella that are unable to remove the bacteria from their processing environment, compromising the entire production line. In this context, an intensive sampling study was conducted to investigate a slaughterhouse with persistent Salmonella problems, establishing the genetic relationship among Salmonella strains isolated during the slaughter process. A total of 36 broiler flocks were sampled during processing at the slaughterhouse. Salmonella was identified based on ISO 6579-1:2017 (Annex D), serotyped by Kauffman-White-Le-Minor technique, and the genetic relationship was assessed with ERIC-PCR followed by PFGE. The outcomes showed that 69.4% of the batches sampled carried Salmonella upon arrival at the slaughterhouse and that 46.3% of the different samples from carcasses were contaminated with Salmonella. The two serovars isolated at the different steps in the slaughterhouse were Enteritidis (98.2%) and Kentucky (1.8%). Pulsed-field gel electrophoresis analysis revealed a low genetic diversity, with all S. Enteritidis isolates showing a nearly identical pulsotype (similarity >85%) and S. Kentucky strains showed the same XbaI PFGE profile (95.0% genetic similarity). The results of this study showed a high genetic relationship among isolates recovered from carcasses and environmental samples in the slaughterhouse from both Salmonella-positive and Salmonella-free flocks. Salmonella strains re-circulated across to poultry flocks and re-entered the slaughterhouse to survive on the processing line. Thus, it is necessary to implement molecular diagnosis methods in time at the field level to determine the Salmonella epidemiology of the flock, to make rapid decisions for the control of Salmonella and prevent entry into the slaughterhouse environment.

Distribution, antimicrobial resistance and genomic characterization of Salmonella along the pork production chain in Jiangsu, China

Salmonella is an important foodborne pathogen that is widely distributed in foods of animal origin. Salmonella strains (n = 246) were identified from 1178 pork samples collected in Jiangsu, China. The most predominant Salmonella serovars in pork and slaughterhouse environment were S. London (49.13%, 85/173) and S. Rissen (53.42%, 39/73), respectively. A total of 13 STs were detected with ST469 dominant (33.33%, 82/246) in these isolates. Most strains were resistant to multiple drugs such as aminoglycosides (88.21%, 217/246), tetracycline (90.24%, 222/246) and florfenicol (91.87%, 226/246). Multiple antimicrobial resistance genes were identified, including that conferring resistance to aminoglycosides (strA, 64/246; strB, 60/246), β-lactam (blaTEM-1, 177/246), phenicols (floR, 115/246) and tetracyclines [tet(A), 216/246; tet(B), 13/246; tet(M), 51/246]. Notably, a novel IncFII(pCoo) blaNDM-1-positive plasmid was characterized. It is worth mentioning that this blaNDM-1-positive Salmonella could be successfully transferred into other Enterobacteriaceae. In addition, phylogenetic analysis indicated a high similarity between blaNDM-1-positive YT2-4-4 and blaNDM-1-positive Salmonella from human sources. To the best of our knowledge, blaNDM-1-carrying Salmonella of pig slaughterhouse origin was first detected. In the present study, we elaborated the distribution, drug resistance, as well as genomic characteristics of Salmonella in the pork production chain, providing valuable baseline data for food safety risk assessment.

Extended-Spectrum ß-Lactamase-Producing Escherichia coli Among Humans, Beef Cattle, and Abattoir Environments in Nigeria.

IntroductionBeef cattle, one of the food-producing animals, are linked to humans through a shared environment and the food chain as a major source of animal protein. Antimicrobial drugs are readily accessible for use in food animal production in Nigeria. Beef cattle and abattoir environments harbor pathogenic bacteria such as Escherichia coli (E. coli) which have developed resistance to antimicrobial agents used for prophylaxis or treatment. This study investigated the zoonotic transmission of extended-spectrum beta-lactamase-producing E. coli (ESBL-EC) among humans, beef cattle, and abattoir environments in Abuja and Lagos, Nigeria.Materials and MethodsWe conducted a cross-sectional study among abattoir workers, beef cattle, and abattoir environments in Abuja and Lagos. Stool, cecal, and environmental samples were collected from apparently healthy workers, slaughtered cattle, and abattoir environments from May to December 2020. Data were collected electronically using open data kit app installed on a mobile phone. Antimicrobial susceptibility patterns were determined using the Kirby–Bauer disk diffusion method against a panel of 16 antimicrobial agents. Phenotypic and genotypic characterizations of the isolates were conducted. Data were analyzed with descriptive statistics.ResultsFrom 21.7% (n = 97) of 448 samples, ESBL-EC were isolated and further characterized. Prevalence of ESBL-EC was highest in cattle (45.4%; n = 44), abattoir workers (41.2%; n = 40), and abattoir environment (13.4%; n = 13). Whole-genome sequencing of ESBL-EC showed dissemination of blaCTX-M-15 (90.7%; n = 88); blaCTX-M-14 (5.2%; n = 5); and blaCTX-M-55 (2.1%; n = 2) genes. The blaCTX-M-15 coexisted with blaCTX-M-14 and blaTEM-1 genes in 2.1% (n = 2) and 39.2% (n = 38) of the isolates, respectively. The presence of blaCTX-M-14 and blaCTX-M-15 genes was significantly associated with isolates originating from abattoir workers when compared with beef cattle isolates (p = 0.05; p < 0.01). The most prevalent sequence types (ST) were ST10 (n = 11), ST215 (n = 7), ST4684 (n = 7), and ST2178 (n = 6). ESBL-EC strain (ST205/B1) harbored mcr-1.1 and blaCTX-M15 and was isolated from a worker at Lagos abattoir. In 91 ESBL-EC isolates, 219 mobile genetic elements (MGEs) harbored resistance genes out of which β-lactam genes were carried on 64 different MGEs. Isolates showed equal distribution of insertion sequences and miniature inverted repeats although only a few composite transposons were detected (humans n = 12; cattle n = 9; environment n = 4). Two isolates of human and cattle origin (ST46/A) harboring ESBL genes and carried by MGEs were clonally related.ConclusionsThis is the first report of blaCTX-M-55 gene in humans and cattle in Nigeria. This study demonstrates the horizontal transfer of ESBL genes possibly by MGEs and buttresses the importance of genomic surveillance. Healthcare workers should be sensitized that people working closely with cattle or in abattoir environments are a high-risk group for fecal carriage of ESBL-EC when compared with the general population.

Escherichia coli serogroups in slaughterhouses: Antibiotic susceptibility and molecular typing of isolates

This study aimed to investigate the contamination of carcasses and slaughterhouse environment with Escherichia coli O157:H7 and non-O157 serogroups (O45:H2, O103:H2, O121:H19, O145:H28, O26:H11, O111:H8). For this purpose, a total of 150 samples (30 carcasses, 30 shredding units, 30 knives, 30 slaughterhouse waste water and 30 wall surfaces) were collected from 5 different slaughterhouses in Kayseri, Turkey. The conventional and molecular methods were performed in order to detect Escherichia coli and its serogroups. Of the 150 samples, 55 (36%) were found to be contaminated with E. coli. Among isolates, E. coli serogroup (O157:H7) were detected in 2 (11%) carcass and 2 (11%) wastewater samples. None of the E. coli isolates harbored tested genes (stx1, stx2, eaeA, and hylA). Effective infection control measures and antibiotic stewardship programs should be adopted to limit the spread of multidrug-resistant bacteria. It was also deduced that these isolates resistance to different antibiotics could be hazardous for public health.

Whole-genome sequencing and gene sharing network analysis powered by machine learning identifies antibiotic resistance sharing between animals, humans and environment in livestock farming.

Anthropogenic environments such as those created by intensive farming of livestock, have been proposed to provide ideal selection pressure for the emergence of antimicrobial-resistant Escherichia coli bacteria and antimicrobial resistance genes (ARGs) and spread to humans. Here, we performed a longitudinal study in a large-scale commercial poultry farm in China, collecting E. coli isolates from both farm and slaughterhouse; targeting animals, carcasses, workers and their households and environment. By using whole-genome phylogenetic analysis and network analysis based on single nucleotide polymorphisms (SNPs), we found highly interrelated non-pathogenic and pathogenic E. coli strains with phylogenetic intermixing, and a high prevalence of shared multidrug resistance profiles amongst livestock, human and environment. Through an original data processing pipeline which combines omics, machine learning, gene sharing network and mobile genetic elements analysis, we investigated the resistance to 26 different antimicrobials and identified 361 genes associated to antimicrobial resistance (AMR) phenotypes; 58 of these were known AMR-associated genes and 35 were associated to multidrug resistance. We uncovered an extensive network of genes, correlated to AMR phenotypes, shared among livestock, humans, farm and slaughterhouse environments. We also found several human, livestock and environmental isolates sharing closely related mobile genetic elements carrying ARGs across host species and environments. In a scenario where no consensus exists on how antibiotic use in the livestock may affect antibiotic resistance in the human population, our findings provide novel insights into the broader epidemiology of antimicrobial resistance in livestock farming. Moreover, our original data analysis method has the potential to uncover AMR transmission pathways when applied to the study of other pathogens active in other anthropogenic environments characterised by complex interconnections between host species.

Distribution and Characteristics of Listeria spp. in Pigs and Pork Production Chains in Germany.

Listeria (L.) monocytogenes is a foodborne pathogen that can cause disease, mainly in elderly, pregnant or immunocompromised persons through consumption of contaminated food, including pork products. It is widespread in the environment and can also be found in asymptomatic carrier animals, for example, in different tissues of pigs. To learn more about their nature, 16 Listeria spp. isolates found in tonsils and intestinal content of pigs and 13 isolates from the slaughterhouse environment were characterized using next-generation sequencing (NGS). A wide distribution of clonal complexes was observed in pigs, as well as in the pork production chain, suggesting multiple sources of entry. Hypervirulent clones were found in pig tonsils, showing the potential risk of pigs as source of isolates causing human disease. The presence of closely related isolates along the production chain suggests a cross-contamination in the slaughterhouse or recontamination from the same source, strengthening the importance of efficient cleaning and disinfection procedures. The phenotypical antimicrobial resistance status of L. monocytogenes isolates was examined via broth microdilution and revealed a low resistance level. Nevertheless, genotypical resistance data suggested multiple resistances in some non-pathogenic L. innocua isolates from pig samples, which might pose a risk of spreading resistances to pathogenic species.

Assessment of Hygienic Status of Some Local Abattoirs and its Impact on Meat Quality

The current study was conducted to evaluate the hygienic status of some local abattoirs and its impact on meat quality through determination of aerobic plate, Enterobacteriaceae, coliforms, Staphylococcus aureus, Enterococci, molds, and yeasts counts. This study was conducted in 5 municipal slaughterhouses located in different provinces, Egypt. A total of 300 samples were collected as following: slaughtered meat, water, and air samples beside swabs from floor, wall, and workers hand (50 samples/each). Statistical analytical results of aerobic plate count clarified that the highest mean value was recorded in the floor swabs (8.9×105 CFU/g) followed by hand swabs of workers (7.41×105 CFU/g) then wall swabs and air samples (7.2×105 CFU/g) then water samples (5.2×105 CFU/g) and lastly meat samples (1.3×105 CFU/g). The highest mean value of Enterobacteriaceae count was recorded in the floor swabs (3.2×104 CFU/g) followed by wall swabs (9.3×103 CFU/g) then air samples (6.63×103 CFU/g) then hand swabs of workers (5.33×103 CFU/g) then water samples (3.19×103 CFU/g) and lastly meat samples (9.02×102 CFU/g). The highest mean value of coliforms count was recorded in the floor swabs (1.65×103 CFU/g) followed by hand swabs of workers (7.23×102 CFU/g) then meat samples (6.10×102 CFU/g) then water samples (4.10×102 CFU/g) then wall swabs (1.72×102 CFU/g) and lastly air samples (1.32×102 CFU/g). On comparison of various microbiological counts on the investigated meat samples with the Egyptian standards, it was clear that most of samples failed to comply with standards confirming the role of contaminated abattoir environment in the obtained result. Results of microbiological examination of the collected samples reflected a clear state of contamination in abattoirs environment that would affect the microbiological quality of the produced meat from these abattoirs that in turn would be harmful for human health.

Evidence of sheep and abattoir environment as important reservoirs of multidrug resistant Salmonella and extended-spectrum beta-lactamase Escherichia coli

The increase in antimicrobial-resistant (AMR) foodborne pathogens, including E. coli and Salmonella in animals, humans, and the environment, is a growing public health concern. Among animals, cattle, pigs, and chicken are reservoirs of these pathogens worldwide. There is a knowledge gap on the prevalence and AMR of foodborne pathogens in small ruminants (i.e., sheep and goats). This study investigates the prevalence and antimicrobial resistance of extended-spectrum beta-lactamase (ESBL) E. coli and Salmonella from sheep and their abattoir environment in North Carolina. We conducted a year-round serial cross-sectional study and collected a total of 1128 samples from sheep (n = 780) and their abattoir environment (n = 348). Sheep samples consisted of feces, cecal contents, carcass swabs, and abattoir resting area feces. Environmental samples consisted of soil samples, lairage swab, animal feed, and drinking water for animals. We used CHROMAgar EEC with 4 μg/ml of Cefotaxime for isolating ESBL E. coli, and ESBL production was confirmed by double-disk diffusion test. Salmonella was isolated and confirmed using standard methods. All of the confirmed isolates were tested against a panel of 14 antimicrobials to elucidate susceptibility profiles. The prevalence of ESBL E. coli and Salmonella was significantly higher in environmental samples (47.7% and 65.5%) compared to the sheep samples (19.5% and 17.9%), respectively (P < 0.0001). We recovered 318 ESBL E. coli and 368 Salmonella isolates from sheep and environmental samples. More than 97% (310/318) of ESBL E. coli were multidrug-resistant (MDR; resistant to ≥3 classes of antimicrobials). Most Salmonella isolates (77.2%, 284/368) were pansusceptible, and 10.1% (37/368) were MDR. We identified a total of 24 different Salmonella serotypes by whole genome sequencing (WGS). The most common serotypes were Agona (19.8%), Typhimurium (16.2%), Cannstatt (13.2%), Reading (13.2%), and Anatum (9.6%). Prevalence and percent resistance of ESBL E. coli and Salmonella isolates varied significantly by season and sample type (P < 0.0001). The co-existence of ESBL E. coli in the same sample was associated with increased percent resistance of Salmonella to Ampicillin, Chloramphenicol, Sulfisoxazole, Streptomycin, and Tetracycline. We presumed that the abattoir environment might have played a great role in the persistence and dissemination of resistant bacteria to sheep as they arrive at the abattoir. In conclusion, our study reaffirms that sheep and their abattoir environment act as important reservoirs of AMR ESBL E. coli and MDR Salmonella in the U.S. Further studies are required to determine associated public health risks.

Isolation and characterization of Staphylococcus argenteus strains from retail foods and slaughterhouses in Japan

Staphylococcus argenteus has been recently established as a novel species of Staphylococcus aureus complex. It is known to cause various human diseases, such as skin and soft-tissue infections, sepsis, and staphylococcal food poisoning, although the source of infection has not been clearly described. In food poisoning cases, the source of bacterial contamination in food is unknown. This study examined the prevalence of S. argenteus among retail fresh food and poultry slaughterhouses in Japan. Among 642 food samples examined, successful isolation of S. argenteus was achieved in 21 of 151 (13.9%) chicken samples. No isolations from pork, beef, fish, or vegetables in retail markets were confirmed. Multiple-locus sequence typing revealed that the 21 isolates were classified into four sequence types (ST) that were divided into 14 subtypes using spa-typing. All food isolates were susceptible to methicillin and did not show positivity for the Panton-Valentine leukocidin gene. When bacteria were isolated from two poultry slaughterhouses in the same region, 14 S. argenteus strains were successfully isolated from only one slaughterhouse. Thirteen of 14 strains were isolated from a poultry carcass and slaughterhouse environments during a certain sampling period and were all classified as ST5961 with identical spa-type. Also, the number of single nucleotide variants (SNVs) detected on the core genomes of the same 13 strains were between 0 and 17, suggesting a long-term inhabitation of an S. argenteus strain inside the facility. Furthermore, one isolate from chicken meat was also genetically linked with the same lineage of slaughterhouse isolates, with ≤15 SNVs being detected. Additionally, one slaughterhouse isolate from chiller water and three chicken isolates were classified into the same cluster by phylogenetic analysis, although the number of pairwise SNVs ranged from 62 to 128. To the best of the authors' knowledge, this is the first study that demonstrated S. argenteus in a food processing facility and the possible bacterial contamination on food during food processing.