Antibiotic Resistant Coliforms Research Articles

Environmental drivers and spatial patterns of antibiotic resistant enteric coliforms across a forest-urban riverscape

Environmental drivers and spatial patterns of antibiotic resistant enteric coliforms across a forest-urban riverscape

On composite sampling for monitoring generic and antibiotic-resistant coliforms in irrigation ponds

On composite sampling for monitoring generic and antibiotic-resistant coliforms in irrigation ponds

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.

Spring of Warta River as a Source of Antibiotic-Resistant Coliform Bacteria

The aim of the study was to evaluate the occurrence of integron-carrying coliform bacteria in the spring water of Warta river, a major river in Poland. We isolated 18 strains (12 Escherichia coli, three Pantoea sp., 2 Citrobacter freundii, and 1 Klebsiella oxytoca) harbouring class 1 and/or class 2 integrons. Class 1 integrons contained aadA1, dfrA7, dfrA17, dfrA1-aadA1, and dfrA17-aadA5 gene cassette arrays, whereas the genetic content of class 2 integrons was stable and consisted of sat2-aad1 gene cassette array. The strains carrying integrons were resistant to 4–11 antimicrobials, most frequently to sulfamethoxazole, ampicillin, piperacillin, trimethoprim, and trimethoprim/sulfamethoxazole.

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 β-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.

Biochar – An efficient sorption material for the removal of pharmaceutically active compounds, DNA and RNA fragments from wastewater

Wastewaters are considered a remarkable source of micropollutants capable of influencing the environment both directly and indirectly. Here we tested porous ecological carbon (Biochar), an effective sorbent material for removing pharmaceuticals, drugs, and their metabolites found in wastewaters. The tested Biochar type was first characterised and used for adsorption experiments of selected micropollutants from a municipal WWTP (wastewater treatment plant) effluent sample. The sorption efficiency was studied on selected pharmaceuticals due to their common presence in aquatic ecosystems. The results show that the studied Biochar type removed the pharmaceuticals with high efficiency (above 90%), so this material can potentially be applied in wastewater treatment. We achieved greater than 99% efficiency in total RNA removal from wastewater. Wastewater might contain infectious RNA fragments of the SARS-CoV-2 virus. However, Biochar can be used as a sorbent in wastewater treatment to remove antibiotic resistance genes. We have also observed a total DNA removal ability of Biochar. On the other hand, the total number and antibiotic-resistant coliform bacteria and enterococci were not changed after Biochar wastewater treatment.

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.

Cross-comparison of methods for quantifying antibiotic resistance in agricultural soils amended with dairy manure and compost

Agricultural soils are often amended with livestock manure, making them a key reservoir of antibiotic resistance genes (ARGs). Given that soils are among the most microbially-diverse environments on the planet; effective characterization and quantification of the effects of manure-derived amendments on soil resistomes is a major challenge. This study examined the effects of dairy manure-derived amendments on agricultural soils via two strategies: quantification of anthropogenic ARG markers via qPCR and shotgun metagenomic resistome profiling; and these strategies were compared to a previously published antibiotic resistant fecal coliform dataset. Soil samples were collected throughout a 120 day complete block field experiment to compare the effects of amendment type on antibiotic resistance. Results of all three measurements were consistent with the hypothesis that the application of composted manure reduced antibiotic resistance in soil relative to the application of raw manure, although some differences were noted in comparing the patterns of the three measurements with time. Raw dairy manure-amended soils yielded high sul1 and tet(W) relative abundances on Day 0 (following amendment application), but significantly decreased to background levels by Day 67 (harvest) and Day 120 (study completion). Shotgun metagenomics similarly detected a decrease in the relative abundances of sulfonamide and tetracycline-associated ARGs over time in the raw manure- and compost-amended soils; however, these levels were significantly lower than those estimated by qPCR. Interestingly, although patterns of sulfonamide and tetracycline resistance among culturable fecal coliforms echoed those observed via qPCR and metagenomics; erythromycin resistant coliforms were directly recovered by culture in amended soils, but corresponding ARGs were not detected by qPCR or metagenomics. This study supports both composting and time restrictions as means of reducing the potential for antibiotic resistance in manure to spread via soil application. Results suggest some differences in finer conclusions drawn depending on which antibiotic resistance monitoring target is selected.

Comparison of antibiotic resistance of coliforms and Escherichia coli strains in industrial and antimicrobial-free poultry manure

ABSTRACT This study aimed to compare the prevalence of antibiotic-resistant coliforms and Escherichia coli strains between industrial and antimicrobial-free poultry manure. The mean counts of high-level antibiotic-resistant coliforms were determined using the spread plate technique. Metal and antibiotic resistance of Escherichia coli strains in manure samples was compared using plate dilution and CLSI procedures, respectively. Several multi-drug resistant Escherichia coli strains were screened for plasmids using alkaline lysis method as well. The relative abundance of high-level antibiotic-resistant coliforms was significantly (p < 0.05 and 0.001) higher in industrial manure than in antimicrobial-free manure. Metal analysis indicated that the concentrations of copper (Cu), zinc (Zn), lead (Pb), and cadmium (Cd) in manure samples were significantly (p < 0.05) correlated with the percentage of coliforms resistant to several antibiotics. The number of multidrug-resistant Escherichia coli strains was significantly (p < 0.001) higher in industrial manure (97.4%) than in antimicrobial-free manure (80.7%). Four out of twelve screened Escherichia coli strains harbored plasmids. Considering the high number of drug-resistant isolates in both samples, reductions in co-selecting agents is essential for the effectiveness of antibiotics, along with controlling the activities which cause the spread of resistance.

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 (ČR). 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 Valašské Meziříčí (ČR). 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 β-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.

Paper currency harbours antibiotic-resistant coliform bacteria and integron integrase.

This study was designed to analyse the prevalence of class 1 and class 2 integron integrase genes among antibiotic-resistant coliform bacteria isolated from paper currency circulating in Pakistan. A total of 500 individual currency notes were collected from different food vending sites at Lahore, Pakistan. Bacterial population were identified by biochemical and PCR techniques. Antimicrobial susceptibility testing was performed by disc diffusion assay. The highest bacterial population on currency was found from street vendors and butcher shops. Escherichia coli was found to be the most prevalent coliform bacteria followed by Klebsiella sp. and Enterobacter sp. PCR amplification of antimicrobial resistance gene showed the presence of ampC, blaTEM , blaNDM-1 , qnrA, tet(A) and tet(B) genes among coliform isolates. A total of 47 integron integrase bearing strains of coliform bacteria were analysed. Sequence analysis showed the presence of dfrA1-aadA1, dfrA1, dfrA5, dfrA7, aadA1, aadA4 cassette arrays in class 1 integron and dfrA1-sat2-aadA1 in class 2 integrase genes. Circulating currency was heavily contaminated with antimicrobial-resistant coliform bacteria bearing class 1 and class 2 integron integrase genes. This study describes a potential threat of severe bacterial infections due to improper hand hygiene and community sanitation when dealing with the currency notes.

Impact of wave action and rainfall on incidence and antibiotic resistance of total coliforms in Southern California beaches.

Coliforms are important bacterial contamination indicators in recreational waters. Little is known about the antibiotic resistance of coliforms from Southern California beaches. This study examined the numbers of coliforms as well as the incidence of antibiotic-resistant coliforms in beaches with restricted and non-restricted wave action by sampling from the shores of both types of beaches following dry and wet weather. Total coliforms were selected by membrane filtration onto mEndo agar and then enumerated. Randomly selected isolates from each location were screened for resistance to nine classes of antibiotics by disk diffusion, and the multiple antibiotic resistance (MAR) index was calculated. Numbers of total coliforms were significantly higher following rain compared to dry weather. Total coliform numbers were not significantly elevated at non-restricted wave action sites. Restricted wave action sites had a 78.5% increase in MAR index following wet weather compared to dry weather. Resistance to ampicillin was observed in almost 50% of isolates and was not significantly impacted by wave action or weather. Minimum inhibitory concentration testing revealed that many isolates were highly resistant to ampicillin. This study is the first to report on the antibiotic resistance of coliforms found in Southern California beaches and highlights the prevalence of ampicillin resistance.

Assessment of antibiotic resistant coliforms from bioaerosol samples collected above a sewage-polluted river in La Paz, Bolivia.

Antimicrobial resistance is a global health risk, and the presence of resistant bacteria in the environment may be an indicative of fecal pollution. The objective of this study has been to assess the antibiotic resistance of airborne coliforms near a highly impacted urban river that may contain high levels of fecal waste. The pilot study has been located within an Andean river basin, the Choqueyapu River basin, which flows through La Paz city in Bolivia. Bioaerosol samples have been collected using liquid impingement and plated on mTEC agar. Coliforms have been detected within 80% of the air samples. The resistance profiles of coliforms present in 20 air samples have been determined by using a modified Kirby-Bauer disk diffusion test against amoxicillin-clavulanic acid, ciprofloxacin, gentamicin, meropenem, sulfamethoxazole-trimethoprim, and tetracycline, antibiotics commonly used to treat gram-negative infection. Broad patterns of antibiotic resistance have been observed throughout the study, with coliforms from at least one sample exhibiting resistance to each of the tested antibiotics. Resistance to sulfamethoxazole and amoxicillin-clavulanic acid has been the most commonly observed, with coliforms in 73% and 60% of samples which helps to demonstrate resistance to these antibiotics, respectively. This study provides insight into the prevalence of airborne, antibiotic resistant coliforms near concentrated fecal waste streams and this highlights an underappreciated hazard and the potential exposure risk in areas where fecal waste may become aerosolized at any given time.

Antibiotic resistant bacteria in surface waters in Slovakia

Abstract This work deals with the occurrence of selected antibiotic resistant bacteria in Slovak surface waters. Total and antibiotic resistant coliform bacteria, enterococci and coagulase positive staphylococci were determined in 34 samples of surface water from rivers and still waters (dams, natural lakes, thermal bath) some of them used for recreational swimming. Sampling was performed during summer season 2017 (July and August). The number of total coliforms ranged from 1.74 log CFU/10 mL to 3.69 log CFU/10 mL. Coliform bacteria were registered in each tested sample. The highest number was observed in the sample from river Hron. Majority of samples contained ampicillin and tetracycline resistant bacteria. Enterococci were determined in 70 % of samples ranging from 0.78 log CFU/10 mL to 3.81 log CFU/10 mL. The highest number of enterococci was observed in river Váh. Ampicillin and vancomycin resistance prevailed over ciprofloxacin and gentamicin resistance. Coagulase positive staphylococci were present in 91 % of samples in the range of 0.70—3.03 log CFU/10 mL. Antibiotic resistant coagulase positive staphylococci were observed only in 32 % of samples, predominantly from rivers. In these samples, chloramphenicol resistance predominated. In three samples of water used for recreational purposes, the limit value for enterococci and E. coli according to Slovak legislation (Decree no. 308/2012 Coll.) for the quality of water in natural swimming pools during bathing season was exceeded.

Boron doped diamond electrode – The elimination of psychoactive drugs and resistant bacteria from wastewater

The study reports the results from applying polycrystalline boron doped diamond (BDD) thin films in eliminating water micropollutants and resistant bacteria. The results documented that BDD anodes were able to effectively decrease the concentration of most of the analyzed pharmaceuticals and drugs by more than 51%. In the case of coliforms and staphylococci the efficiency level reached almost 100%. The study investigates the occurrence of 29 selected psychoactive drugs and metabolites in 18 wastewater treatment plants (WWTPs) in the Slovak and Czech Republic. It should be mentioned that the under investigation geographic area has a population of more than 1.5 million inhabitants. The analyses of influents and effluents were performed using in-line SPE-LC-MS/MS. It was found that the most concentrated substances were cotinine, tramadol, methamphetamine and venlafaxine, and compounds such as tramadol, venlafaxine, oxazepam and citalopram and had limited degradation efficiency at all monitored WWTPs with a maximum of 30%. Antibiotic resistant coliforms and staphylococci were also found to be present in the Slovak wastewaters; whereas in Czech effluents, no chloramphenicol and tetracycline resistant E. coli and S. aureus were detected.

Spatial relationships between small-holder farms coupled with livestock management practices are correlated with the distribution of antibiotic resistant bacteria in northern Tanzania

We examined the spatial distribution of antibiotic-resistant coliform bacteria amongst livestock from three distinct cultural groups, where group-level differences in practices (e.g., antibiotic use) may influence the magnitude of antibiotic resistance, while livestock interactions (e.g., mixing herds, shared markets) between these locations may reduce heterogeneity in the distribution of antibiotic resistant bacteria. Data was collected as part of a larger study of antibiotic-resistance in northern Tanzania. Simple regression and generalized linear regression were used to assess livestock management and care practices in relation to the prevalence of multidrug-resistant (MDR) coliform bacteria. Simple and multivariable logistic regression were then used to identify how different management practices affected the odds of households being found within MDR “hotspots.” Households that had a higher median neighbourhood value within a 3000 m radius showed a significant positive correlation with livestock MDR prevalence (β = 4.33, 95% CI: 2.41–6.32). Households were more likely to be found within hotspots if they had taken measures to avoid disease (Adjusted Odds Ratio (AOR) 1.53, CI: 1.08—2.18), and if they reported traveling less than a day to reach the market (AOR 2.66, CI: 1.18—6.01). Hotspot membership was less likely when a greater number of livestock were kept at home (AOR 0.81, CI: 0.69–0.95), if livestock were vaccinated (AOR 0.32, CI: 0.21—0.51), or if distance to nearest village was greater (AOR 0.46, CI: 0.36–0.59). The probability of MDR increases when herds are mixed, consistent with evidence for passive transmission of resistant bacteria between animals. Reduced MDR with vaccination is consistent with many studies showing reduced antibiotic use with less disease burden. The neighbourhood effect has implications for design of intervention studies.

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 (Petržalka) in the capital of Slovakia during 1year. 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.

Occurence of antibiotic resistant bacteria in raw cow milk from vending machines

Abstract The prevalence of antibiotic resistant coliform bacteria and enterococci in fresh raw cow milk obtained from four vending machines in Slovakia was assessed. Also, differences in the number of antibiotic-resistant bacteria in milk according to season (winter and summer) were compared. Number of total coliform bacteria ranged from 2.45 to 4.18 log CFU/mL. Majority of them were resistant to ampicillin. This was expected due to their intrinsic resistance apart from Escherichia coli which is sensitive to ampicillin. In addition, we observed also tetracycline and gentamicin resistance. Each of our samples contained E.coli. Number of total enterococci ranged from 1.95 to 3.78 log CFU/mL. We have observed predominantly vancomycin resistance in all tested samples. In samples taken during winter we have found higher number of total and resistant coliforms as well as total enterococci compared to those taken during summer.

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

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. 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. 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. 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.