Occurrence of Antibiotic-Resistant Bacteria in Fish and Seafood from Slovak Market.

The consumption of sushi or poke has grown globally. However, this type of dish often contains raw fish or seafood; therefore, it can pose a microbial risk for consumers. This study deals with the occurrence of total and antibiotic-resistant coliform bacteria and enterococci in fish and seafood as well as sushi and poke bought from Slovak retail (restaurants and fast food). Total coliforms have ranged in sushi, poke samples and samples of fish and seafood from cooling counters from 0.6 to 5.1 log CFU/g. Ampicillin resistance has been predominantly observed in all types of samples. Tetracycline resistance was detected in 16% of all tested samples and gentamicin resistance in 13%. Total enterococci has been detected in 74% of sushi samples, 100% of poke samples and 62% of samples obtained from supermarkets. The majority of enterococci were resistant to ampicillin. Vancomycin resistance was observed in five samples. Forty-eight resistant coliforms were identified mainly as Enterobacter spp. and Klebsiella spp. Antibiotic-resistant isolates were predominantly resistant to gentamicin, chloramphenicol and tetracycline. In 13% of resistant isolates was detected efflux pumps overproduction, and in four isolates was detected the tetA resistance gene. Our results point to poor hygiene in some establishments. The prevention of the antibiotic-resistant bacteria spread would be in better stewardship and improved monitoring of sanitation.

Occurrence of Antibiotic Resistant Bacteria in Flours and Different Plant Powders Used in Cuisine.

In recent years, several alimentary diseases have been connected with the consumption or tasting of raw flour and dough. Microbiological quality concern is also raising due to increased consumer demand for plant powders, while some of them can be consumed without prior thermal processing. In this study, we have focused on the occurrence of antibiotic-resistant coliform bacteria and enterococci in flour, plant powder and dough from Slovak retail. Our results indicated the presence of both total and antibiotic-resistant coliform bacteria and enterococci in the flour and powder samples. Lower numbers of the total, as well as resistant bacteria, were detected in flours compared to plant powders. Coliform bacteria isolates were predominantly identified as Klebsiella spp. and Enterobacter spp. Ampicillin resistance appeared in 97% of isolates followed by chloramphenicol resistance (22%) and tetracycline resistance (17%). The presence of the blaSHV gene was confirmed in 13% of isolates. The tetA and tetE genes were present in 25% of isolates of coliform bacteria. The presence of enterococci was detected only in plant powders. Antibiotic-resistant strains were identified as the following: Enterococcus casseliflavus, E. gallinarium and E. faecium. Despite the isolates showing resistance to vancomycin, the presence of the vanA gene was not detected. The majority of antibiotic-resistant isolates belonged to the group of medium biofilm producers. None of these isolates showed efflux pump overproduction. Antibiotic-resistant coliform bacteria and enterococci were not detected in the processed doughs.

Characterization of Antibiotic Resistant Coliform Bacteria and Resistance Genes Isolated from Samples of Smoothie Drinks and Raw Milk.

Raw foodstuffs have been marked as a healthier alternative in the context of nutrient content and are becoming more popular with consumers. Thermally untreated foods may represent a microbiological risk connected with the possible presence of antimicrobial resistance. The aim of this study was to prove that popular raw food beverages such as smoothies and raw milk may be a source of antibiotic-resistant coliform bacteria and resistant genes. The majority of antibiotic-resistant isolates (110) were identified as Enterobacter spp., Escherichia coli, and species of Klebsiella spp., predominantly ß-lactam and chloramphenicol resistant. Multidrug resistance has been registered in one-third of resistants. Overproduction of efflux pumps was clarified in 8 different bacteria. The majority of resistant isolates were strong biofilm producers. Antibiotic resistance gene blaOXA was detected in 25% of isolates, especially in E. coli. Resistance genes blaTEM and blaSHV were detected in 19% and 14%, respectively. This is the first study to point out that popular raw drinks such as smoothies or raw milk, besides their nutrient benefits, could represent a reservoir of antibiotic-resistant bacteria as well as antibiotic resistance genes. According to this, raw drinks could contribute to the dissemination of antibiotic resistance in the human gastrointestinal tract and environment.

Microbial Safety of Smoothie Drinks from Fresh Bars Collected in Slovakia.

Among the many consumers in Slovakia, smoothies are nowadays gaining popularity. Smoothie drinks are prepared from raw fruits and vegetables. Therefore, their microbiological safety depends on hygiene standards. The aim of this work was to monitor and quantify selected sensitive and antibiotic-resistant microorganisms present in collected smoothies. Twenty analyzed smoothie samples were collected from six food service establishments (fresh bars) in the capital city of Slovakia, Bratislava. Antibiotic-resistant bacteria were found in at least one of each fresh bar. Antibiotic-resistant coliform bacteria prevailed, especially in green smoothies or juices containing more vegetable ingredients. Resistance to ampicillin, ciprofloxacin, tetracycline, chloramphenicol, and gentamicin was observed in the case of coliform bacteria. More than half of the smoothie drink samples did not contain resistant enterococci. On the other hand, vancomycin-resistant enterococci were detected in 20% of samples. The most frequently isolated antibiotic-resistant strains belonged to the Enterobacter spp. or Klebsiella spp. genus. In the last part of the work, the pretreatment effect of smoothie components on the selected microorganisms' counts in the final product was investigated. Washing ingredients with an aqueous solution of a biocide agent containing silver and hydrogen peroxide proved to be the most effective way to decrease bacterial counts.

Effervescent ferrate(VI)-based tablets as an effective means for removal SARS-CoV-2 RNA, pharmaceuticals and resistant bacteria from wastewater.

Waterborne pathogens including viruses, bacteria and micropollutants secreted from population can spread through the sewerage system. In this study, the efficiency of unique effervescent ferrate-based tablets was evaluated for total RNA and DNA removal, disinfection and degradation of micropollutants in hospital wastewater. For the purpose of testing, proposed tablets (based on citric acid or sodium dihydrogen phosphate) were used for various types of hospital wastewater with specific biological and chemical contamination. Total RNA destruction efficiency using tablets was 70-100% depending on the type of acidic component. DNA destruction efficiency was lower on the level 51-94% depending on the type of acidic component. In addition, our study confirms that effervescent ferrate-based tablets are able to efficiently remove of SARS-CoV-2 RNA from wastewater. Degradation of often detected micropollutants (antiepileptic, antidepressant, antihistamine, hypertensive and their metabolites) was dependent on the type of detected pharmaceuticals and on the acidic component used. Sodium dihydrogen phosphate based tablet appeared to be more effective than citric acid based tablet and removed some pharmaceuticals with efficiency higher than 97%. Last but not least, the disinfection ability was also verified. Tableted ferrates were confirmed to be an effective disinfectant and no resistant microorganisms were observed after treatment. Total and antibiotic resistant bacteria (coliforms and enterococci) were determined by cultivation on diagnostic selective agar growth media.

Hospital Wastewater-Important Source of Multidrug Resistant Coliform Bacteria with ESBL-Production.

This work compares the prevalence of antibiotic resistant coliform bacteria in hospital wastewater effluents in Slovak (SR) and Czech Republic (CR). It also describes selected antibiotic resistant isolates in view of resistance mechanism and virulence factor. The highest number of multidrug resistant bacteria was detected in samples from the hospital in Valasské Mezirící (CR). More than half of resistant isolates showed multidrug resistance phenotype as well as strong ability to form biofilm. In 42% of isolates efflux pump overproduction was detected together with tetA and tetE genes. The production of extended-spectrum ß-lactamases in coliform isolates was encoded mainly by blaTEM, blaCTX-M-2 and blaCTX-M-8/25 genes. About 62% of resistants contained a combination of two or more extended spectrum beta-lactamases (ESBL) genes. Our results strengthen the fact that hospital effluents are a source of multidrug resistant bacteria which can spread their resistance genes to other bacteria in wastewater treatment plants (WWTPs). Accordingly, hospital wastewater should be better treated before it enters urban sewerage.