Sediment samples from three coastal sites - two beach resorts (Beach 1 and Beach 2 sites) and an area lying between an oil refinery and a river estuary (Estuarine site) - were analyzed for antibiotic- and heavy metal (HM)-resistant enterococci.A total of 123 enterococci, 36 E. faecium, 34 E. casseliflavus, 33 E. hirae, 5 E. faecalis, 3 E. durans, 3 E. gallinarum, and 9 Enterococcus spp, were recovered. Strains resistant to erythromycin, tetracycline and quinupristin/dalfopristin (Q/D) were recovered from all sites, whereas multidrug-resistant isolates were recovered only from “Beach 2” (14%) and “Estuarine” (3.7%). As regards HM resistance, the strains showed a high frequency (68%) of cadmium and/or copper resistance and uniform susceptibility to mercury. The prevalence of cadmium-resistant strains was significantly higher among erythromycin-resistant than among erythromycin-susceptible strains. A significant association between cadmium or copper resistance and Q/D resistance was also observed at “Estuarine” site. The levels of the two HMs in sediment from all sites were fairly low, ranging from 0.070 to 0.126 μg/g, for cadmium and from 1.00 to 7.64 μg/g for copper. Mercury was always undetectable. These findings are consistent with reports that low HM concentrations may contribute to co-selection of antibiotic-resistant bacterial strains, including enterococci.

The determination of values of abundance of antibiotic resistance genes (ARGs) per mass of soil is extremely useful to assess the potential impacts of relevant sources of antibiotic resistance, such as irrigation with treated wastewater or manure application. Culture-independent methods and, in particular, quantitative PCR (qPCR), have been regarded as suitable approaches for such a purpose. However, it is arguable if these methods are sensitive enough to measure ARGs abundance at levels that may represent a risk for environmental and human health. This study aimed at demonstrating the range of values of ARGs quantification that can be expected based on currently used procedures of DNA extraction and qPCR analyses. The demonstration was based on the use of soil samples spiked with known amounts of wastewater antibiotic resistant bacteria (ARB) (Enterococcus faecalis, Escherichia coli, Acinetobacter johnsonii, or Pseudomonas aeruginosa), harbouring known ARGs, and also on the calculation of expected values determined based on qPCR.The limits of quantification (LOQ) of the ARGs (vanA, qnrS, blaTEM, blaOXA, blaIMP, blaVIM) were observed to be approximately 4 log-units per gram of soil dry weight, irrespective of the type of soil tested. These values were close to the theoretical LOQ values calculated based on currently used DNA extraction methods and qPCR procedures. The observed LOQ values can be considered extremely high to perform an accurate assessment of the impacts of ARGs discharges in soils. A key message is that ARGs accumulation will be noticeable only at very high doses. The assessment of the impacts of ARGs discharges in soils, of associated risks of propagation and potential transmission to humans, must take into consideration this type of evidence, and avoid the simplistic assumption that no detection corresponds to risk absence.

In developing countries, many urban rivers are suffering from heavy contamination by untreated sewage, which implies great microbial risks. However, information regarding the bacterial pathogen diversity and distribution in urban rivers is highly limited. In this study, 41 water samples of fifteen rivers and eight samples from two sewage treatment plants in Changzhou City of Yangtze River Delta were sampled. Next-generation sequencing and a self-built reference pathogen database were used to investigate the diversity of enteric and environmental pathogens. The results indicated that the studied urban rivers were harboring diverse potential pathogen species, which primarily included enteric pathogens in Arcobacter and Bacteroides, and environmental pathogens in Acinetobacter, Aeromonas and Pseudomonas. Quantification of twelve pathogens/indicators of interest by qPCR showed that Escherichia coli, Enterococcus faecalis, Campylobacter jejuni, Arcobacter cryaerophilus, Acinetobacter johnsonii, Acinetobacter lwoffii and Aeromonas spp. were abundant, with median values ranging from 3.30 to 5.85 log10 copies/100 mL, while Salmonella, Legionella pheumophila, Mycobacterium avium, Pseudomonas aeruginosa and Staphylococcus aureus were infrequently quantified. The pollution of nutrients and human intestinal microorganisms indicated by specific markers were found to be prevalent but with different levels in the rivers. The correlation analyses revealed that the diversity (p < 0.01) and concentrations (p < 0.05) of the enteric pathogens highly correlated to the human fecal marker abundances, which indicated that the enteric pathogens in the urban rivers were likely to have originated from domestic sewage. The environmental pathogens, which are different from the enteric ones, showed various distribution patterns, and some of them were more abundant in the rivers of rich nutrient. Our findings provide a comprehensive understanding of the bacterial pathogen distribution and influencing factors in urban rivers that are impacted by domestic sewage, thereby establishing the foundation for urban water management.

Bats are poorly understood as a reservoir of multidrug-resistant strains; therefore, the aim of this study was to determine molecular characterization of multidrug-resistant Enterococcus strains isolated from bat species from Poland. A multi-stage analysis based on targeted isolation of drug-resistant strains (selective media with tetracycline, chloramphenicol, gentamicin, streptomycin, and vancomycin), determination of the phenotypic profile of drug-susceptibility using the disc diffusion method, and amplification of DNA fragments surrounding rare restriction sites (ADSRRS-fingerprinting) was used for the isolation and differentiation of strains. The applied strategy finally allowed identification of E. faecalis resistant to at least one antimicrobial in 47.2% of the single-animal group and in 46.9% of the pooled samples of bat’s guano. Out of the 36 distinct isolates, 69% met the criteria of multi-drug resistance, with a dominant combination of resistance to tetracycline, erythromycin, and rifampicin. Simultaneously, 41.6% of the strains were high-level aminoglycoside resistant (HLAR). In most strains, phenotypic resistance was reflected in the presence of at least one gene encoding resistance to a given drug. Moreover, our research results show that some genes were detected simultaneously in the same strain statistically significantly more frequently. This may confirm that the spread of some genes (tetM and ermB or aph (3′)-IIIa as well as gelE and aac (6′)-Ie-aph (2″)-Ia or ant (6)-Ia) is associated with their common occurrence on the same mobile genetic element.To our knowledge, this is the first analysis of multidrug-resistance among E. faecalis isolated from bats. Our research demonstrates that the One Health concept is not associated exclusively with food-producing animals and humans, but other species of wildlife animals should be covered by monitoring programs as well.We confirmed for the first time that bats are an important reservoir of multi-resistant E. faecalis strains and could have a great impact on environmental resistance.

The existence of micropollutants (MPs) including pathogens in waters poses great risks to ecological safety and human health. Sulfate radical (SO4•−)‒based advanced oxidation processes (AOPs) have attracted considerable attention in water treatment for both disinfection and removal of emerging MPs. Here, we investigated the SO4•−‒mediated kinetic and mechanistic aspects of simultaneous inactivation of Enterococcus faecalis (E. faecalis) and degradation of carbamazepine (CBZ), a typical MP with high occurrence in wastewater. In the absence of CBZ, (73.8 ± 2.3) % E. faecalis were inactivated after 12 min of treatment, while in the presence of CBZ, (68 ± 1.6) % of E. faecalis were inactivated, exhibiting similar inactivation efficiency with or without MP. The second‒order rate constant (k) of E. faecalis reacting with SO4•− was determined to be (5.42 ± 0.64) × 10⁹ M⁻¹ s⁻¹. In addition, two distinct types of disinfection models, one based on the quenching ratio (Q model) and the other on steady‒state concentration of SO4•− (R model), were developed to predict the inactivation kinetics of E. faecalis. Both models exhibited good performance for describing the disinfection of E. faecalis with RMSE of 0.065 and 0.048, respectively. Our kinetic experimental and modelling results on bacterial disinfection and degradation of CBZ were projected to offer valuable insight into future developments for typical wastewater scenarios where microorganisms and MPs coexist.

To examine the influence of anthropogenic activities on the marine ecosystem around the Andaman and Nicobar Islands, a study was conducted to investigate the abundance of heterotrophic, indicator and pathogenic bacteria during the monsoon season. We noticed the higher abundance of heterotrophic, indicator (Escherichia coli and Enterococcus faecalis) and pathogenic bacterial counts (Aeromonas hydrophila, Enterobacter aerogens, Escherichia coli strain O157:H7, Shigella dysenteriae and Vibrio parahaemolyticus) near the 10ᵒ channel, which is the principal route to reach Andaman mainland. Most of the stations are offshore (chosen to cover shipping tracks) at a distance ranging from 41 to 266 km from the coast. The nearest station to the coast was at 21 km away. Apart from those mentioned above, several other pathogenic bacteria were also detected such as Klebsiella pneumoniae, Salmonella paratyphi, Vibrio cholera and Vibrio vulnificus but they are sparsely detected at few stations only.

This study was performed to evaluate the abundance and diversity of Enterococcus sp. and the distribution of biomarker genes in Enterococcus faecalis in Port Blair Bay, Andaman and Nicobar Islands. The Enterococcus sp. densities at the seven sampling stations were highly influenced by tidal fluctuations and season. The distributions and diversities of species varied in the inner and outer regions of Port Blair Bay. Among the 1816 total isolates, the occurrence of fecal Enterococcus was high (1.78×104CFU/100mL) in Phoenix Bay. Moreover, 67.76% of the isolates were identified as Enterococcus, and the most frequently identified species were E. hirae, E. avium and E. faecalis. Assessments of antibiotic resistance and biomarker genes revealed the maximum occurrence in the Aberdeen Bay isolates. The most prevalent biomarker genes observed in the E. faecalis isolates were gelE and asa1, whereas cyl was not found among the isolates. In silico sequence analysis of biomarker genes of E. faecalis also revealed that they are evolutionarily well conserved with those of earlier reports. Further, multivariate analysis distinguished the JB, PB and OS stations from the other stations according to distinctive microbial densities and compositions. In addition, the Shannon-Wiener diversity indices and box-whisker plots further facilitated and supported the multivariate results.

Sewage is one of the biggest contributors to coastal pollution. The study was aimed to assess the impact of sewage on coastal water quality of Kanyakumari, the southernmost part of India. A bacteriological survey was made on distribution and abundance of fecal indicators and human pathogenic bacteria and seasonal influence on the bacterial load and antibiotic resistance of the isolates. Samples were collected from sewage discharge point along the eastern shore of Kanyakumari Coast from February 2019 to January 2020. Nine pollution indicator bacteria and pathogenic species such as Escherichia coli, Klebsiella spp., Enterococcus faecalis, Aeromonas spp., Proteus mirabilis, Salmonella typhi, Vibrio cholerae, Shigella spp. and Flavobacterium spp. were isolated from the samples. These isolates were tested against 10 antibiotics, using Kirby Bauer method. All the isolates were resistant to at least two antibiotics. The presence of antibiotic resistant bacteria has been used as bio-indicators of pollution. Hence it is clear that the domestic sewage entering the coast is untreated which might lead a serious impact on human and marine wildlife along coastlines.

Microbiological sea water quality is a public health problem that has serious repercussions in the tourism and economy of Colombia. This study determines the concentrations of Escherichia coli, Enterococcus faecalis and Clostridium perfringens at eleven beach water points and seven streams along the coast of the Department of Atlántico, Colombia. In seawater, total E. coli, E. faecalis and C. perfringens concentrations were found between 16 and 572 cfu/100 mL, 7–450 cfu/100 ml and 2–125 cfu/100 ml, respectively. The highest counts were observed mainly on urbanised beaches and in correspondence with streams whose waters had a high concentration of faecal origin microorganisms, which represent a serious health risk factor for bathers. Relevant efforts have to be addressed to improve the microbiological quality of these beaches by the establishment of efficient wastewater management programs aimed at enhancing the efficiency of the local treatment plant and the control of illegal sewage pouring onto the coast.

In order to examine the health of the coastal waters off Visakhapatnam in terms of prevalence and abundance of heterotrophic (H), indicator and pathogenic (P) bacterial counts (BC) and influence of physical processes on them, time-series observations were conducted during January (winter), March (spring), July (summer) and October (post-monsoon). We noticed the impact of physical forces on substantial variations in abundance and distribution of the HBC, total coliforms, Enterococcus faecalis and Pseudomonas aeruginosa in the study region. Based on our results Escherichia coli and other PBC were not much influenced by the physical conditions. It has been noticed that the perennial existence of the high abundance of IBC and PBC above the standard limits during the entire study period leading to an alarming situation in the coastal waters off Visakhapatnam.