First study in the frequency of isolation and phenotypic antimicrobial resistance profiles of pig and cattle origin Campylobacter strains in Romania.

Campylobacter spp. is recognized as one of the most common pathogens involved in the development of gastrointestinal infections in humans. The current study aimed to enhance the knowledge on the occurrence and molecular characterization of Campylobacter spp. in pigs and cattle origin caecum samples (n = 56) collected in one year, from nine Romanian slaughterhouses, and to determine the antimicrobial resistance profile of the isolated strains. All Campylobacter spp. strains (n = 41) isolated from swine and cattle caecum samples were analyzed in terms of antimicrobial resistance, in accordance with the EURL protocol and with the Commission Implementing Decision No. 2020/1729. The prevalence rate for C. coli. in pig caecum samples was 92.3% (36/39), and the prevalence of C. jejuni, in cattle origin samples was 29.4% (5/17). C. coli strains isolated from pigs proved resistant to tetracycline 75% (27/36), ciprofloxacin 69.4% (25/36), erythromycin 8.3% (3/36), ertapenem 2.8% (1/36) and gentamicin 2.8% (1/36), but no resistance was observed towards chloramphenicol. C. jejuni strains originating from cattle expressed resistance to ciprofloxacin 60.0% (3/5) and tetracycline 20.0% (1/5), but they were susceptible to chloramphenicol, erythromycin, ertapenem and gentamicin. In the present study, 19.5% (7/36) C. coli strains isolated from pigs were identified as multidrug-resistant (MDR) bacteria. The obtained results demonstrated that pigs especially, but cattle to, can be considered important natural reservoirs for zoonotic multidrug-resistant Campylobacter strains, having a stimulating effect for further studies aiming at the molecular screening of the genotypic antimicrobial resistance processing of a higher number of samples.

Causes of Post-Mortem Carcass and Organ Condemnations and Economic Loss Assessment in a Cattle Slaughterhouse.

The study was undertaken to investigate the main causes of carcass and organ condemnations, as well as to estimate the financial losses suffered by a cattle slaughterhouse. In this regard, an active abattoir survey, based on standard post-mortem inspection procedures for meat, was conducted on 151,741 cattle, from January 2021 to December 2022. Overall, 13.27% (n = 20,125) of the carcasses expressed lesions or pathological conditions and, out of them, 1.15% (n = 1738) were totally confiscated, while another 12.12% (n = 18,387) were partially admitted for human consumption. In the case of organs, the general inspection data reveal that 12.28% (n = 18,630), 7.56% (n = 11,477), 1.89% (n = 2862), and 0.27% (n = 412) of the examined liver, lung, heart, and kidney specimens presented one or more types of abnormalities. In addition, regarding the types of specific pathological findings, dystrophies/anomalies (69.8%), circulatory disorders (40.6%), fecal contamination (60.9%), and suspected bacterial/viral infections showed a dominant occurrence in the liver, lung, heart, and kidneys, respectively. Consequently, the total direct financial losses resulting from edible part condemnation over the two years was estimated at EUR 4,021,717.3, which represents 1.17% of the total achievable net revenue without carcass and organ condemnation. Of this, EUR 3,661,400.4 (1.07%) and EUR 360,316.9 (8.73%) was related to carcass and organ condemnation, respectively. The study results demonstrate that the post-mortem inspection of meat at the slaughterhouse level plays a crucial role in identifying pathological lesions, in addition to some other issues, such as fecal contamination or non-compliant laboratory results, relevant to both public health and economic factors.

Evaluation of Antimicrobial Resistance Profiles of Bacteria Isolated from Biofilm in Meat Processing Units.

The aim of this study was to assess the hygiene of pork, beef, and poultry carcasses and to determine the phenotypic antibiotic susceptibility of the bacteria embedded in the biofilm formed on the carcasses kept in cooling chambers for at least three days. The level of hygiene was assessed by determining the total aerobic colony count (TACC) and the Enterobacteriaceae level in different sampling points of the carcasses, along with the detection of E. coli and Pseudomonas spp. embedded in the biofilm. Furthermore, the E. coli and Pseudomonas spp. isolates were tested for antimicrobial resistance profiles. A total of 130 samples collected from pork, beef, and poultry from processing units were analyzed to determine the total aerobic colony count as well as to measure the level of Enterobacteriaceae found on the carcasses. The antimicrobial susceptibility of 44 Escherichia coli and eight Pseudomonas spp. strains isolated from the carcasses were assessed using the Vitek 2 system using two different cards. Overall, the regulatory limits for the TACC were exceeded in 7.6% of the samples, and 65% of the samples exceeded the regulatory limits for Enterobacteriaceae levels. The antimicrobial susceptibility tests of the E. coli isolates analyzed with the AST-GN27 card revealed the highest resistance to be that towards ampicillin (76.1%), followed by cefazolin (71.4%), amoxicillin/clavulanic acid (61.9%), nitrofurantoin (52.3%), cefoxitin (47.6%), tetracycline (38.1%), piperacillin, norfloxacin (19%), trimethoprim-sulfamethoxazole (11.9%), cefotaxime (9.5%), ceftazidime, cefazolin, amikacin, gentamicin, and ciprofloxacin (4.7%). However, all of the isolates were sensitive to piperacillin-tazobactam and imipenem. Thirty-two (61.5%; 95% CI 47.9-73.5) out of fifty-two isolates exhibited multidrug resistance, resulting in the expression of 10 resistance profiles. The findings of this study highlight serious hygienic and sanitary deficiencies within the meat processing units and demonstrate that the resulting meat can harbor Multidrug-resistant Escherichia coli and Pseudomonas spp., both of which pose a serious public health risk. However, further research with a larger number of samples is required to reach thorough results.

Occurrence of Campylobacter spp. and Phenotypic Antimicrobial Resistance Profiles of Campylobacter jejuni in Slaughtered Broiler Chickens in North-Western Romania

Campylobacteriosis is recognized as one of the most common food-borne zoonoses, with worldwide distribution, having undercooked poultry meat and other cross-contaminated foodstuffs as the main sources of human infections. The current study aimed to provide data on the occurrence of the thermophilic Campylobacter spp. in seven broiler chicken flocks, from three north-western Transylvanian counties of Romania, as well as to determine the antimicrobial resistance profile of the isolated C. jejuni strains. A total of 324 fresh cecal samples were collected during the slaughtering process, and screened for the presence of Campylobacter spp., using routine microbiological and molecular diagnostic tools. Overall, 85.2% (276/324; 95% CI 80.9-88.6) of the tested samples expressed positive results for Campylobacter spp., with dominant occurrence of C. coli towards C. jejuni (63.4% vs. 36.6%). From the six tested antimicrobials, the 101 isolated C. jejuni strains were resistant against ciprofloxacin (79.2%), nalidixic acid (78.2%), tetracycline (49.5%), and streptomycin (7.9%), but total susceptibility was noticed against erythromycin and gentamicin. Seven (6.9%) isolates exhibited multidrug resistance. The study results emphasize the role of broiler chicken as reservoir of Campylobacter infections for humans, as well as strengthen the necessity of the prudent using of antimicrobials in the poultry industry.

Occurrence, Pathogenic Potential and Antimicrobial Resistance of Escherichia coli Isolated from Raw Milk Cheese Commercialized in Banat Region, Romania.

The aim of the present study was to investigate the presence, pathogenic potential and antimicrobial susceptibility profile of Escherichia coli isolated from raw milk cheese, traditionally produced by farmers and marketed directly to the consumer in Banat region, Romania. A total of 81.1% (43/53) of the processed samples expressed positive results for E. coli, with a distribution of 83.8% (31/37), and 75.0% (12/16) in the cow- and sheep-milk-origin assortments, respectively. Overall, 69.8% (30/43) of the specimens had a contamination level ≤10 CFU/g. Molecular tests showed that, from the total number of E. coli isolates, 9.3% (4/43) harbored the stx2, and 2.3% (1/43), the stx1 virulence genes. The E. coli O157 (including H7) biovariety was identified in 7% (3/43) of the samples by the Vidas equipment. From the 27 antimicrobials tested with the Vitek2 automated system, the E. coli isolates displayed resistance to enrofloxacin (100%, 15 out of 15 tested isolates), ampicillin (39.5%, 17/43), norfloxacin (28.6%, 8/28), fosfomycin (25%, 7/28), amoxicillin/clavulanic acid (23.3%, 10/43), cefalexin (20%, 3/15), cefalotin (13.3%, 2/15), tetracycline (13.3%, 2/15), trimethoprim-sulfamethoxazole (9.3%, 4/43), piperacillin-tazobactam (7.1%, 2/28), cefotaxime (7.1%, 2/28), cefepime (7.1%, 2/28), ticarcillin/clavulanic acid (6.7%, 1/15), florfenicol (6.7%, 1/15), ceftazidime (3.6%, 1/28), and ertapenem (3.6%, 1/28). Ten (23.3%) strains were multidrug-resistant. The obtained preliminary results indicated hygienic-sanitary deficiencies throughout the cheese production process, and demonstrated that these products can harbor virulent and multidrug-resistant E. coli strains, which constitute a public health risk. However, future investigations, processing a higher number of samples, are still necessary to draw comprehensive conclusions.

Diversity and Antibiotic Resistance Profiles of Listeria monocytogenes Serogroups in Different Food Products from the Transylvania Region of Central Romania.

ABSTRACT: This study used molecular serogrouping to assess the presence and antimicrobial susceptibility profile of Listeria monocytogenes isolates from food products of different animal origin, collected from a county situated in the historical region of Transylvania in central Romania. Seventeen (7.7%) of 221 screened samples were positive for L. monocytogenes; these included 8 (6.2%) of 130 ready-to-eat products (i.e., sausages, ham, and smoked specialties), 6 (12.8%) of 47 raw meat samples (i.e., minced pork, pork organs, and snails), and 3 (6.8%) of 44 dairy samples (i.e., assortment of cheeses). The identified L. monocytogenes serogroups were 1/2a-3a (47.1%), 4b-4d-4e (29.4%), 1/2c-3c (11.8%), and 4a-4c (11.8%). All isolates were resistant to benzylpenicillin and fusidic acid. Resistance was also detected toward oxacillin (88.2%), fosfomycin (82.4%), clindamycin (76.5%), imipenem (52.9%), ciprofloxacin (41.2%), rifampin (41.2%), trimethoprim-sulfamethoxazole (29.4%), and tetracycline (29.4%). On the other hand, all isolates proved susceptible to gentamicin, moxifloxacin, teicoplanin, vancomycin, tigecycline, erythromycin, and linezolid. All tested strains exhibited multidrug resistance, resulting in the expression of a total of 12 resistance profiles. These findings extend the understanding of the spread of an important pathogen in Romanian food products, highlighting a substantial public health issue and medical concern, especially for consumers with a compromised health status.

Multidrug Resistant Coagulase-Positive Staphylococcus aureus and Their Enterotoxins Detection in Traditional Cheeses Marketed in Banat Region, Romania.

The main objectives of the present study were to determine the occurrence of coagulase positive staphylococci (CPS) and to assess the presence and antimicrobial susceptibility profile of Staphylococcus aureus isolates in different raw milk origin (cow and sheep) traditional cheeses marketed in Banat region, Romania. Additionally, the presence of mecA gene in S. aureus isolates and the staphylococcal enterotoxins (SEs) in cheese samples were evaluated. A total of 81.6% (138/169) of the screened samples were positive for CPS. Furthermore, 35.5% (49/138) of the investigated CPS positive cheese samples were contaminated with S. aureus, with an isolation frequency of 46.6% (14/30) in caș, 33.3% (32/96) in telemea, 25% (2/8) in burduf, and 25% (1/4) in urda assortments, respectively. From the total number of S. aureus isolates, 6.1% (3/49) harbored the mecA gene. Detectable levels of SEs were identified in 4.3% (4/94) of cheese samples with a CPS contamination level higher than 105 log CFU g-1. The expressed antimicrobial susceptibility profile of the tested cheese-origin S. aureus isolates, with the automated Vitek 2 equipment, showed resistance towards amikacin (90.1%, 10 out from 11 tested), enrofloxacin (86.2%, 25/29), ceftiofur (72.7%, 8/11), neomycin (63.6%, 7/11), benzylpenicillin (53.1%, 26/49), kanamycin (41.4%, 12/29), rifampicin (39.5%, 15/38), tetracycline (38.8%, 19/49), tilmicosin (36.4%, 4/11), clindamycin (30.6%, 15/49), ciprofloxacin (30%, 6/20), erythromycin (22.4%, 11/49), tylosin (18.2%, 2/11), oxacillin (16.3%, 8/49), linezolid (15%, 3/20), teicoplanin (15%, 3/20), fusidic acid (13.1%), imipenem (10.5%, 4/38), vancomycin (7.9%, 3/38), ampicillin (5.5%, 1/18), mupirocin (5.5%, 1/18), fosfomycin (5%, 1/20), and gentamicin (4.1%, 2/49). Twenty-four (49%) S. aureus isolates exhibited multidrug resistance. The investigation highlighted a common occurrence of multidrug-resistant S. aureus strains in the monitored cheese assortments, which can constitute a potential risk for consumers' health.