Selected Cardiovascular Risk Markers in Vegetarians and Subjects of General Population.

OBJECTIVE: Besides genetic factors there are environmental effects including nutritional habits which can influence the risk of age-related diseases. The aim of the study was to assess the age dependence of selected cardiovascular risk markers in two groups of subjects with different nutritional pattern. METHODS: In 470 long-term vegetarians and 478 subjects of general population the following indicators were measured: total cholesterol, HDL-cholesterol, triacylglycerol, glucose, insulin concentrations, LDL-cholesterol, atherogenic index and insulin resistance IR(HOMA) were also calculated in studied subjects. Obtained data were evaluated according to age decades. RESULTS: Vegetarian vs. non-vegetarian concentrations of total cholesterol, LDL-cholesterol, insulin, and values of atherogenic index and IR(HOMA) were significantly reduced in all age decades. Vegetarian vs. non-vegetarian triacalglycerol concentrations were significantly reduced from 4th­7th decade. Vegetarian average decade values of all lipid parameters were in reference range. In non-vegetarian group, the risk average values of total cholesterol (>5.2 mmol/l) were found from 5th­7th decade, LDL-cholesterol (>3.3 mmol/l) in 7th decade and atherogenic index (>4) in 6th­7th decade. In vegetarians vs. non-vegetarians were noted the average decade values for total cholesterol ranging from 4.01­4.59 vs. 4.48­5.67 mmol/l, for triacylglycerols 1.00­1.33 vs. 1.13­1.74 mmol/l, for LDL-cholesterol 2.03­2.58 vs. 2.43­3.49 mmol/l, for atherogenic index 2.72­3.31 vs. 3.05­4.21 and for IR(HOMA) 0.99­1.15 vs. 1.15­1.84. CONCLUSION: Our data show significantly reduced mean age decade values of lipid and non-lipid cardiovascular risk markers in all adult vegetarians. Smaller changes of markers between decades compared to non-vegetarians document a protective effect of vegetarian nutrition in prevention of cardiovascular disease.

Properties of anaerobic fungi isolated from several habitats: complexity of phenotypes.

Isolates of anaerobic fungi from rumen, animal faeces and compost displayed morphological similarity with known anaerobic fungi. According to their ITS sequences, species were related to Neocallimastix and Piromyces. Rumen fungi tolerated exposure to an aerobic atmosphere for at least four days. Under anaerobic conditions, they could grow on both, defined or complex substrates. Growth in liquid media was monitored by the continuous measurement of metabolic gases (O2, CO2, H2, CO, H2S, CH4). Monitored metabolism was complex, showed that both CO2 and H2 were produced and subsequently consumed by yet unknown metabolic pathway(s). CO and H2S were evolved similarly, but not identically with the generation of CO2 and H2 suggesting their connection with energetic metabolism. Anaerobic fungi from snail faeces and compost produced concentrations of H2S, H2, CO near the lower limit of detection. The rumen isolates produced cellulases and xylanases with similar pH and temperature optima. Proteolytic enzymes were secreted as well. Activities of some enzymes of the main catabolic pathways were found in cell-free homogenates of mycelia. The results indicate the presence of the pentose cycle, the glyoxylate cycle and an incomplete citrate cycle in these fungi. Differences between isolates indicate phenotypic variability between anaerobic fungi.

Wastewater analysis: the mean of the monitoring of frequently prescribed pharmaceuticals in Slovakia.

The estimation of medication use is based on the statistical data from pharmacies and hospitals. Excessive use or misuse of some compounds, especially psychoactive medications, has not yet been monitored in Slovakia. Wastewater analysis provides useful data about the medication use and misuse in individual regions. This study is focused on the analysis of 23 substances in the wastewaters of Slovakia. The monitoring programme has included stimulants, opioid and morphine derivatives, benzodiazepines, antidepressants, drug precursors and their metabolites. Urinary markers of these compounds were analysed at WWTP influent in seven regions (Bratislava, Kosice, Zvolen, Banská Bystrica, Trencín, Presov a Piestany) using LC-MS/MS technique. The analysis was performed from March to October 2013. The pattern in use of these compounds was also monitored. Tramadol and venlafaxine were found to be the most concentrated compounds among of all studied psychoactive pharmaceuticals. The highest specific loads of tramadol were detected in Piestany (409 mg/day/1000 inhabitants) and Zvolen (366 mg/day/1000 inhabitants). There is a considerable number of spa facilities (hotels) situated in these cities and this fact contributes to a higher occurrence of these psychoactive compounds in respective wastewaters.

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.

Microplastics in the Food Chain.

Currently, microplastics represent a widespread contamination found in almost every part of the environment. The plastic industry has generated waste since the 1950s, which unfortunately now counts in the millions. The largest share of plastic consumption is used to produce packaging materials, including those applied in the food industry. The versatility of plastic materials is mainly due to their lightness, flexibility, strength, and persistence. Although plastic materials are widely used due to their beneficial properties, contamination of the environment with microplastics and nanoplastics is an emerging problem worldwide. This type of contamination is endangering animal life and thus also the food chain and public health. This review summarizes the knowledge about microplastics in the food chain. The effect of microplastics on the food chain has been particularly studied in marine organisms, and research deals less with other food commodities. Therefore, based on the studied literature, we can conclude that the issue is still not sufficiently examined, and should be paid more attention to maintain the health of the population.

Hospital Wastewater-Source of Specific Micropollutants, Antibiotic-Resistant Microorganisms, Viruses, and Their Elimination.

Municipal wastewaters can generally provide real-time information on drug consumption, the incidence of specific diseases, or establish exposure to certain agents and determine some lifestyle consequences. From this point of view, wastewater-based epidemiology represents a modern diagnostic tool for describing the health status of a certain part of the population in a specific region. Hospital wastewater is a complex mixture of pharmaceuticals, illegal drugs, and their metabolites as well as different susceptible and antibiotic-resistant microorganisms, including viruses. Many studies pointed out that wastewater from healthcare facilities (including hospital wastewater), significantly contributes to higher loads of micropollutants, including bacteria and viruses, in municipal wastewater. In addition, such a mixture can increase the selective pressure on bacteria, thus contributing to the development and dissemination of antimicrobial resistance. Because many pharmaceuticals, drugs, and microorganisms can pass through wastewater treatment plants without any significant change in their structure and toxicity and enter surface waters, treatment technologies need to be improved. This short review summarizes the recent knowledge from studies on micropollutants, pathogens, antibiotic-resistant bacteria, and viruses (including SARS-CoV-2) in wastewater from healthcare facilities. It also proposes several possibilities for improving the wastewater treatment process in terms of efficiency as well as economy.

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.

Non-antimicrobial pharmaceuticals can affect the development of antibiotic resistance in hospital wastewater.

Within this study, we were interested in the effects of wastewater on the development of antimicrobial resistance. Microorganisms can relatively promptly adapt to evolutionary pressures of the environment, including antibiotics. Therefore, we tested how the adaptability of the model microorganism Salmonella enterica is affected by wastewater full of pharmaceuticals, illicit drugs, and other micropollutants. Wastewater samples had been taken from effluent of hospitals and from wastewater treatment plant (WWTP) Petrzalka influent and effluent. In these samples, presence of 38 substances was monitored. The highest concentration was observed in case of tramadol, citalopram, venlafaxine, cotinine, atenolol, valsartan, carbamazepine, azithromycin, and ciprofloxacin. According to this data, we focused also on individual pharmaceutical compounds presented in wastewater samples in elevated concentrations. Effect on resistance development of two pain relief medications (carbamazepine, tramadol), hypotensive medications (atenolol, valsartan), and the nicotine metabolite (cotinine) was also investigated. For this study, we employed concentrations presented in wastewater as well as in urine of patients and/or users. To determine the frequency of mutations leading to ciprofloxacin resistance, we applied the modified Ames test employing the strain Salmonella Typhimurium. Resistance index increased in the case of all wastewater samples from conventional hospitals where we observed a 1.22-1.69-fold increase of mutations leading to ciprofloxacin resistance. Tested compounds caused rise of resistance index in lower concentrations found in wastewater. The most significant increase of resistance index was detected after carbamazepine treatment.

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.

Development of triclosan and antibiotic resistance in Salmonella enterica serovar Typhimurium.

The possible association between the use of triclosan and the development of antibiotic resistance was examined in triclosan-resistant mutants of Salmonella enterica serovar Typhimurium. These mutants were obtained from a sensitive parental strain and from ciprofloxacin-resistant isogenic strains using spontaneous mutagenesis or selection after one short exposure or continuous exposure to low concentrations of triclosan. The results showed that triclosan in the environment does not increase the mutation frequency but selects bacterial strains with reduced antibiotic susceptibility. This property depended on the multiple antibiotic resistance (Mar) phenotype of bacterial strains and on the triclosan concentration.

Annual changes in the occurrence of antibiotic-resistant coliform bacteria and enterococci in municipal wastewater.

Wastewater contains subinhibitory concentrations of different micropollutants such as antibiotics that create selective pressure on bacteria. This phenomenon is also caused by insufficient wastewater treatment technology leading to the development and spread of antibiotic-resistant bacteria and resistance genes into the environment. Therefore, this work focused on monitoring of antibiotic-resistant coliform bacteria and enterococci in influent and effluent wastewaters taken from the second biggest wastewater treatment plant (Petrzalka) in the capital of Slovakia during 1 year. Antibiotic-resistant strains were isolated, identified, and characterized in terms of susceptibility and biofilm production. All of 27 antibiotic-resistant isolates were identified mainly as Morganella morganii, Citrobacter spp., and E. coli. Multidrug-resistance was detected in 58% of isolated strains. All tested isolates could form biofilm; two strains were very strong producers, and 74% formed biofilm by strong intensity. The flow rate of the influent wastewater had a more significant impact on the number of studied bacteria than the temperature. Graphical abstract.

Hospital wastewaters treatment: Fenton reaction vs. BDDE vs. ferrate(VI).

Various types of micropollutants, e.g., pharmaceuticals and their metabolites and resistant strains of pathogenic microorganisms, are usually found in hospital wastewaters. The aim of this paper was to study the presence of 74 frequently used pharmaceuticals, legal and illegal drugs, and antibiotic-resistant bacteria in 5 hospital wastewaters in Slovakia and Czechia and to compare the efficiency of several advanced oxidations processes (AOPs) for sanitation and treatment of such highly polluted wastewaters. The occurrence of micropollutants and antibiotic-resistant bacteria was investigated by in-line SPE-LC-MS/MS technique and cultivation on antibiotic and antibiotic-free selective diagnostic media, respectively. The highest maximum concentrations were found for cotinine (6700 ng/L), bisoprolol (5200 ng/L), metoprolol (2600 ng/L), tramadol (2400 ng/L), sulfamethoxazole (1500 ng/L), and ranitidine (1400 ng/L). In the second part of the study, different advanced oxidation processes, modified Fenton reaction, ferrate(VI), and oxidation by boron-doped diamond electrode were tested in order to eliminate the abovementioned pollutants. Obtained results indicate that the modified Fenton reaction and application of boron-doped diamond electrode were able to eliminate almost the whole spectrum of selected micropollutants with efficiency higher than 90%. All studied methods achieved complete removal of the antibiotic-resistant bacteria present in hospital wastewaters.

Prevalence of antibiotic-resistant coliform bacteria, Enterococcus spp. and Staphylococcus spp. in wastewater sewerage biofilm.

OBJECTIVES: Urban wastewater contains various micropollutants and a high number of different micro-organisms. Some bacteria in wastewater can attach to surfaces and form biofilm, which gives bacteria an advantage in the fight against environmental stresses. This work focused on analysis of bacterial communities in biofilms isolated from influent and effluent sewerage of a wastewater treatment plant (WWTP) in Bratislava, Slovakia. METHODS: Detection of biofilm microbiota was performed by culture-independent and -dependent approaches. The composition of bacterial strains was detected by denaturing gradient gel electrophoresis fingerprinting coupled with construction of 16S rRNA clone libraries. Analysis of the concentration of antibiotics and the prevalence of antibiotic-resistant coliforms, Enterococcus spp. and Staphylococcus spp. in sewerage was also studied. RESULTS: Biofilm collected at the inlet point was characterised primarily by the presence of Pseudomonas spp., Acinetobacter spp. and Janthinobacterium spp. clones, whilst members of the genus Pseudomonas were largely detected in biofilm isolated in outflow of the WWTP. Predominant antibiotics such as azithromycin, clarithromycin and ciprofloxacin were found in influent wastewater. The removal efficiency of these antibiotics, notably azithromycin and clarithromycin, was 30% in most cases. CONCLUSION: The highest number of antibiotic-resistant bacteria, with a predominance of coliforms, was detected in samples of effluent biofilm. Multidrug-resistant strains in effluent biofilm showed very good biofilm-forming ability.

Phenolic acids from plant foods can increase or decrease the mutation frequency to antibiotic resistance.

Phenolic compounds are ubiquitous in plant foods, and they have been reported to have multiple biological effects. This study investigates the ability of derivatives of hydroxybenzoic and hydroxycinnamic acids to influence the development of ciprofloxacin resistance in the antibiotic-sensitive strain Salmonella enterica subsp. enterica serovar Typhimurium. We have found that cinnamic acid and its hydroxy derivatives increase the frequency of mutations leading to ciprofloxacin resistance in comparison with spontaneous mutagenesis. Derivatives of hydroxybenzoic acid showed no or very little effect. Interestingly, gallic acid caused only phenotype resistance but not arising of mutant strains. Vanillic acid decreased spontaneous mutation frequency, leading to resistance not only to ciprofloxacin but also to tetracycline and gentamicin, and also decreased mutation frequency induced by 3-(5-nitro-2-furyl)acrylic acid, sodium azide, and hydrogen peroxide leading to ciprofloxacin resistance.

Zerovalent iron and iron(VI): Effective means for the removal of psychoactive pharmaceuticals and illicit drugs from wastewaters.

Herein we report the analysis of 27 selected psychoactive compounds found in the wastewater of the largest suburb in the eastern part of Central Europe Bratislava­Petrzalka, Slovakia. Thirteen of them (MDMA, methamphetamine, amphetamine, THC-COOH, benzoylecgonine, codeine, tramadol, venlafaxine, oxazepam, citalopram, methadone, EDDP, cocaine) were found in concentrations above 30 ng/L. These compoundswere selected for further monitoring. The possibility of complete degradation of these 13 substances by zerovalent iron and iron(VI) was studied in thewastewater from the Petrzalka treatment plant. During the week the concentration of themajority of the studied compounds inwastewaterwas stable. Concentrations of MDMA, cocaine, tramadol, and oxazepam reached significantly higher levels during the weekend.Only about 10% removal efficiency for tramadol, venlafaxine, oxazepam, MDMA, citalopram, methadone, and EDDP was observed at the treatment plant. In contrast, methamphetamine, amphetamine, and codeine were removed with 68%, 83%, and 53% efficiency, respectively. The degradation of synthetic drugs (methamphetamine, cocaine, MDMA) in wastewater is limited, while cannabis (of natural biological origin) is degradedwith efficiency greater than 90%. After utilization of the Fenton reaction, its modification, and use of ferrate(VI), a high efficiency of eliminating all of these substances to values below the limit of detection was achieved.

Development of ciprofloxacin resistance due to chromosomal mutations induced by 2-nitrofluorene.

In this paper we investigated the ability of 2-nitrofluorene to induce mutations leading to antibiotic resistance in quinolone-sensitive strain Salmonella typhimurium. After preincubation of bacteria with 2-nitrofluorene, the frequency of mutation to ciprofloxacin resistance was 57 fold higher than in the case of spontaneous mutability. Some of resultant resistant colonies showed a great increase of ciprofloxacin MIC.

The effect of phenolic and polyphenolic compounds on the development of drug resistance.

The effect of two phenolic compounds vanillin (4-hydroxy-3-methoxybenzaldehyde) and lignin on the development of drug/antibiotic resistance in Salmonella typhimurium was studied. Using the modified Ames test we have shown that vanillin alone has negligible effect on spontaneous mutability to ciprofloxacin and gentamicin resistance. At the tested concentrations vanillin reduces the toxicity of 4-nitroquinoline-N-oxide (4NQO) and reduces the ability of this compound to induce mutations leading to ciprofloxacin but not to gentamicin resistance. Lignin at higher concentrations increases mutagenicity to ciprofloxacin resistance and possess considerable inhibition effect on the spontaneous and 4NQO induced mutability to gentamicin resistance.