Wastewater surveillance of antibiotic resistance and class 1 integron-integrase genes: Potential impact of wastewater characteristics on genes profile.

Antibiotic resistance (AR) is a major global health concern, but current surveillance efforts primarily focus on healthcare settings, leaving a lack of understanding about AR across all sectors of the One Health approach. To bridge this gap, wastewater surveillance provides a cost-effective and efficient method for monitoring AR within a population. In this study, we implemented a surveillance program by monitoring the wastewater effluent from two large-scale municipal treatment plants situated in Isfahan, a central region of Iran. These treatment plants covered distinct catchment regions and served a combined population about two million of residents. Furthermore, the effect of physicochemical and microbial characteristics of wastewater effluent including biological oxygen demand (BOD), chemical oxygen demand (COD), total suspended solids (TSS), temperature, total coliforms and Escherichia coli concentration on the abundance of ARGs (blaCTX-M, tetW, sul1, cmlA, and ermB) and class 1 integron-integrase gene (intI1) were investigated. Sul1 and blaCTX-M were the most and least abundant ARGs in the two WWTPs, respectively. Principal Component Analysis showed that in both of the WWTPs all ARGs and intI1 gene abundance were positively correlated with effluent temperature, but all other effluent characteristics (BOD, COD, TSS, total coliforms and E. coli) showed no significant correlation with ARGs abundance. Temperature could affect the performance of conventional activated sludge process, which in turn could affect the abundance of ARGs. The results of this study suggest that other factors than BOD, COD and TSS may affect the ARGs abundance. The predicted AR could lead to development of effective interventions and policies to combat AR in the clinical settings. However, further research is needed to determine the relationship between the AR in wastewater and clinical settings as well as the effect of other influential factors on ARGs abundance.

The bacterial biofilm resistome in drinking water distribution systems: A systematic review.

Antibiotic resistance in drinking water systems poses human health risks. Earlier studies, including reviews on antibiotic resistance in drinking water systems are limited to the occurrence, behaviour and fate in bulk raw water and drinking water treatment systems. By comparison, reviews on the bacterial biofilm resistome in drinking water distribution systems are still limited. Therefore, the present systematic review investigates the occurrence, behaviour and fate and, detection methods of bacterial biofilm resistome in the drinking water distribution systems. A total of 12 original articles drawn from 10 countries were retrieved and analyzed. Antibiotic resistant bacteria and antibiotic resistance genes detected in biofilms include those for sulfonamides, tetracycline, and beta-lactamase. The genera detected in biofilms include Staphylococcus, Enterococcus, Pseudomonas, Ralstonia, Mycobacteria, as well as Enterobacteriaceae family and other gram-negative bacteria. The presence of Enterococcus faecium, Staphylococcusaureus, Klebsiella pneumoniae, Acinetobacterbaumannii, Pseudomonas aeruginosa, and Enterobacter species (ESKAPE bacteria) among the detected bacteria points to potential human exposure and health risks especially for susceptible individuals via the consumption of drinking water. Besides, the effects of water quality parameter and residual chlorine, the physico-chemical factors controlling the emergence, persistence and fate of the biofilm resistome are still poorly understood. Culture-based methods, and molecular methods, and their advantages and limitations are discussed. The limited data on the bacterial biofilm resistome in drinking water distribution system points to the need for further research. To this end, future research directions are discussed including understanding the formation, behaviour, and fate of the resistome and the controlling factors.

Microbial characteristics of municipal solid waste compost: Occupational and public health risks from surface applied compost.

Land application of MSW compost promotes the soil fertility and productivity, but there is concern about its chemical and microbial quality. Although, current microbial indicators of MSW compost are generally considered to be protective of human health, the use of these indicators doesn't adequately represent the presence of pathogens that might be more resistant to inactivation during composting process. This work aims evaluation of the microbial characteristics of MSW compost and estimation of the health risks associated with exposure to pathogens of potential concern in MSW compost. Although the recommended standards for fecal coliforms and Salmonella were met in a relatively high percentage of samples, there were detectable levels of microbial pathogens. The highest daily infection risk was estimated for Cryptosporidium (1.25 × 10-3 per person) followed by Salmonella and Campylobacter, while the lowest related to adenovirus. Infection risks were low for both farmers and children in one-time exposure and all pathogen risks were decreased with increasing waiting time to near insignificant levels. However, the median annual risk of cryptosporidiosis or gastroenteritis was above the recommended value of 10-3 per person per year. Because of potential presence of pathogens in MSW compost, some level of pathogen monitoring beyond conventional indicators may be required to estimate potential risks. The findings of this study could provide information to governments and local authorities in making decisions on measures to reduce risk from application of MSW compost. Further research is needed to obtain the reliable information about the inactivation of microorganisms in environment.

The air-borne antibiotic resistome: Occurrence, health risks, and future directions.

Antibiotic resistance comprising of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) is an emerging problem causing global human health risks. Several reviews exist on antibiotic resistance in various environmental compartments excluding the air-borne resistome. An increasing body of recent evidence exists on the air-borne resistome comprising of antibiotic resistance in air-borne bioaerosols from various environmental compartments. However, a comprehensive review on the sources, dissemination, behavior, fate, and human exposure and health risks of the air-borne resistome is still lacking. Therefore, the current review uses the source-pathway-receptor-impact-mitigation framework to investigate the air-borne resistome. The nature and sources of antibiotic resistance in the air-borne resistome are discussed. The dissemination pathways, and environmental and anthropogenic drivers accounting for the transfer of antibiotic resistance from sources to the receptors are highlighted. The human exposure and health risks posed by air-borne resistome are presented. A health risk assessment and mitigation strategy is discussed. Finally, future research directions including key knowledge gaps are summarized.

Tracking antibiotic resistance genes and class 1 integrons in Escherichia coli isolates from wastewater and agricultural fields.

Considering high concentrations of multidrug-resistant bacteria and antibiotic resistance genes (ARGs) in wastewater, agricultural reuse of treated wastewater may be a public health threat due to ARG dissemination in different environmental compartments, including soil and edible parts of crops. We investigated the presence of antibiotic-resistant Escherichia coli as an indicator bacterium from secondary treated wastewater (STWW), water- or wastewater-irrigated soil and crop samples. ARGs including blaCTX-m-32, blaOXA-23, tet-W, sul1, cml-A, erm-B, along with intI1 gene in E. coli isolates were detected via molecular methods. The most prevalent ARGs in 78 E. coli isolates were sul1 (42%), followed by blaCTX-m-32 (19%), and erm-B (17%). IntI1 as a class 1 integrons gene was detected in 46% of the isolates. Cml-A was detected in STWW isolates but no E. coli isolate from wastewater-irrigated soil and crop samples contained this gene. The results also showed no detection of E. coli in water-irrigated soil and crop samples. Statistical analysis showed a correlation between sul1 and cml-A with intI1. The results suggest that agricultural reuse of wastewater may contribute to the transmission of antibiotic-resistant bacteria to soil and crop. Further research is needed to determine the potential risk of ARB associated with the consumption of wastewater-irrigated crops.

Antibiotic resistance and class 1 integron genes distribution in irrigation water-soil-crop continuum as a function of irrigation water sources.

The increasing demand for fresh water coupled with the need to recycle water and nutrients has witnessed a global increase in wastewater irrigation. However, the development of antibiotic resistance hotspots in different environmental compartments, as a result of wastewater reuse is becoming a global health concern. The effect of irrigation water sources (wastewater, surface water, fresh water) on the presence and abundance of antibiotic resistance genes (ARGs) (blaCTX-m-32, tet-W, sul1, cml-A, and erm-B) and class 1 integrons (intI1) were investigated in the irrigation water-soil-crop continuum using quantitative real-time PCR (qPCR). Sul1 and blaCTX-m-32 were the most and least abundant ARGs in three environments, respectively. The abundance of ARGs and intI1 significantly decreased from wastewater to surface water and then fresh water. However, irrigation water sources had no significant effect on the abundance of ARGs and intI1 in soil and crop samples. Principal component analysis (PCA) showed that UV index and air temperature attenuate the abundance of ARGs and intI1 in crop samples whereas the air humidity and soil electrical conductivity (EC) promotes the ARGs and intI1. So that the climate condition of semi-arid regions significantly affects the abundance of ARGs and intI1 in crop samples. The results suggest that treated wastewater might be safely reused in agricultural practice in semi-arid regions without a significant increase of potential health risks associated with ARGs transfer to the food chain. However, further research is needed for understanding and managing ARGs transfer from the agricultural ecosystem to humans through the food chain.

COVID-19 infection risk from exposure to aerosols of wastewater treatment plants.

Fecal shedding of SARS-CoV-2 from COVID-19 patients and presence of the viral RNA in wastewater have extensively been reported. Some wastewater treatment plant (WWTP) processes generate aerosols which have the potential to transmit pathogenic microorganisms and present a health risk for exposed individuals. We analyzed the presence of viral RNA of SARS-CoV-2 in raw wastewater and air samples of WWTPs. The risk that may arise from exposure to virus-contaminated aerosols of wastewater was estimated by developing a quantitative microbial risk analysis (QMRA) method. SARS-CoV-2 was detected in 9 of 24 (37.5%) wastewater samples with a concentration about 104 genomic copies L-1. The viral RNA was also detected in 40% (6/15) of air samples. QMRA analysis showed a relatively high risk of SARS-CoV-2 infection for wastewater workers via exposure to the viral aerosols. The estimated annual infection risk ranged from 1.1 × 10-2 to 2.3 × 10-2 per person per year (PPPY) for wastewater workers which was higher than the reference level recommended by WHO (10-3 pppy). However, due to the lack of data on survival of SARS-CoV-2 in wastewater and its fate in aerosolized state, more research is needed to determine the importance of wastewater in transmission of COVID-19.

Detection of antibiotic resistant Acinetobacter baumannii in various hospital environments: potential sources for transmission of Acinetobacter infections.

BACKGROUND: Antibiotic resistant Acinetobacter baumannii has emerged as one of the most problematic hospital acquired pathogens around the world. This study was designed to investigate the presence of antibiotic resistant A. baumannii in various hospital environments. METHODS: Air, water and inanimate surface samples were taken in different wards of four hospitals and analyzed for the presence of A. baumannii. Confirmed A. baumannii isolates were analyzed for antimicrobial susceptibility and also screened for the presence of three most common OXA- type carbapenemase-encoding genes. RESULTS: A. baumannii was detected in 11% (7/64) of air samples with the highest recovery in intensive care units (ICUs). A. baumannii was also detected in 17% (7/42) and 2% (1/42) of surface and water samples, respectively. A total of 40 A. baumannii isolates were recovered and analysis of antimicrobial susceptibility showed the highest resistance towards ceftazidime (92.5%, 37/40). 85% (34/40) and 80% (32/40) of the isolates were also resistant to imipenem and gentamicin, respectively. Resistance genes analysis showed that 77.5% (31/40) strains contained OXA-23 and 5% (2/40) strains contained OXA-24, but OXA-58 was not detected in any of the strains. CONCLUSION: Detection of antibiotic resistant A. baumannii in various samples revealed that hospital environments could act as a potential source for transmission of A. baumannii infections especially in ICUs. These results emphasize the importance of early detection and implementation of control measures to prevent the spread of A. baumannii in hospital environments.

Hospital air: A potential route for transmission of infections caused by ß-lactam-resistant bacteria.

BACKGROUND: The emergence of bacterial resistance to ß-lactam antibiotics seriously challenges the treatment of various nosocomial infections. This study was designed to investigate the presence of ß-lactam-resistant bacteria (BLRB) in hospital air. METHODS: A total of 64 air samples were collected in 4 hospital wards. Detection of airborne bacteria was carried out using culture plates with and without ß-lactams. BLRB isolates were screened for the presence of 5 common ß-lactamase-encoding genes. Sequence analysis of predominant BLRB was also performed. RESULTS: The prevalence of BLRB ranged between 3% and 34%. Oxacillin-resistant bacteria had the highest prevalence, followed by ceftazidime- and cefazolin-resistant bacteria. The frequency of ß-lactamase-encoding genes in isolated BLRB ranged between 0% and 47%, with the highest and lowest detection for OXA-23 and CTX-m-32, respectively. MecA had a relatively high frequency in surgery wards and operating theaters, whereas the frequency of blaTEM was higher in intensive care units and internal medicine wards. OXA-51 was detected in 4 wards. Acinetobacter spp, Acinetobacter baumannii, and Staphylococcus spp were the most predominant BLRB. CONCLUSIONS: The results revealed that hospital air is a potential route of transmission of BLRB, such as Acinetobacter and Staphylococcus, 2 important causative agents of nosocomial infections. Therefore, improvement of control measures against the spreading of airborne bacteria in hospital environments is warranted.