Distribution and Relationships of Antimicrobial Resistance Determinants among Extended-Spectrum-Cephalosporin-Resistant or Carbapenem-Resistant Escherichia coli Isolates from Rivers and Sewage Treatment Plants in India.

To determine the distribution and relationship of antimicrobial resistance determinants among extended-spectrum-cephalosporin (ESC)-resistant or carbapenem-resistant Escherichia coli isolates from the aquatic environment in India, water samples were collected from rivers or sewage treatment plants in five Indian states. A total of 446 E. coli isolates were randomly obtained. Resistance to ESC and/or carbapenem was observed in 169 (37.9%) E. coli isolates, which were further analyzed. These isolates showed resistance to numerous antimicrobials; more than half of the isolates exhibited resistance to eight or more antimicrobials. The blaNDM gene was detected in 14/21 carbapenem-resistant E. coli isolates: blaNDM-1 in 2 isolates, blaNDM-5 in 7 isolates, and blaNDM-7 in 5 isolates. The blaCTX-M gene was detected in 112 isolates (66.3%): blaCTX-M-15 in 108 isolates and blaCTX-M-55 in 4 isolates. We extracted 49 plasmids from selected isolates, and their whole-genome sequences were determined. Fifty resistance genes were detected, and 11 different combinations of replicon types were observed among the 49 plasmids. The network analysis results suggested that the plasmids sharing replicon types tended to form a community, which is based on the predicted gene similarity among the plasmids. Four communities each containing from 4 to 17 plasmids were observed. Three of the four communities contained plasmids detected in different Indian states, suggesting that the interstate dissemination of ancestor plasmids has already occurred. Comparison of the DNA sequences of the blaNDM-positive plasmids detected in this study with known sequences of related plasmids suggested that various mutation events facilitated the evolution of the plasmids and that plasmids with similar genetic backgrounds have widely disseminated in India.

Microminipigs as a new experimental animal model for toxicological studies: comparative pharmacokinetics of perfluoroalkyl acids.

In this study, we evaluated the efficacy of a novel minipig strain, the Microminipig (MMPig), as an animal model for studying the pharmacokinetics of a mixture of 10 perfluoroalkyl acids (PFAAs). After a single oral dose was given, we found that the blood depuration of PFAAs (blood t1/2), which we calculated using first-order elimination curves, ranged from 1.6 to 86.6 days. Among the five body compartments analyzed, the liver was the greatest site of accumulation of perfluorooctanesulfonate and longer chain perfluorinated carboxylates such as perfluorodecanoic acid, perfluoroundecanoic acid and perfluorododecanoic acid. We observed an increasing accumulation trend of perfluorinated carboxylates in the organs associated with the fluorinated carbon chain length. The perfluorononanoic acid burden was the highest among the treated compounds 21 days after a single exposure, as 29% of the given perfluorononanoic acid dose was accumulated in the tissues. The persistence of PFAAs in edible pig tissues even after 21 days post-exposure raises concerns about the safety of swine products. This was the first study to use MMPigs to elucidate the pharmacokinetics of a group of environmental pollutants. We found that MMPigs could be excellent experimental animals for toxicological studies due to their easy handling, cost efficacy for target compounds and ease of waste treatment.

Spatio-temporal distribution, source, and genotoxic potential of polycyclic aromatic hydrocarbons in estuarine and riverine sediments from southern India.

In order to categorize the distribution, source, and effects of polycyclic aromatic hydrocarbons (PAHs) in aquatic systems of southern India, chemical and toxicological analyses were performed on surface and core sediments, collected from Adyar river, Cooum river, Ennore estuary, and Pulicat lake near Chennai city. The total PAH concentration in surface sediment ranged from 13 to 31,425ng/g with a mean value of 4320ng/g; the concentration was markedly higher in Cooum river compared to that at other sites. The historical PAH dissemination in core samples in the Cooum river, Ennore estuary, and Pulicat lake ranged from 30 to 31,425ng/g, from 8.6 to 910ng/g, and from 62 to 546ng/g, respectively. Surface sediments were predominantly contaminated with low molecular weight (LMW) PAHs. Historical profiles suggest that PAH contamination in the area is now greater than it had been in the past. PAH accumulation in Pulicat lake was distinct from that at other locations where high molecular weight (HMW) PAHs were predominant. DNA damage in HepG2 cells treated with sediment extracts from different locations showed a good correlation with their respective total PAH levels. Statistical analysis revealed that 3-ring and 4-ring PAHs may synergistically contribute to the genotoxic potency compared to others in sediments. The study also showed that a majority of PAHs in the study area indicated a petrogenic origin. Based on the enrichment and toxicological assessment of PAHs in sediments, Cooum river was shown to suffer the highest biological impairment among the studied water bodies.

Has irrigated water from Mahaweli River contributed to the kidney disease of uncertain etiology in the dry zone of Sri Lanka?

The Mahaweli is the largest river basin in Sri Lanka that provides water to the dry zone region through multipurpose irrigation schemes . Selenium, arsenic, cadmium, and other bioimportant trace elements in surface waters of the upper Mahaweli River were measured using ICP-MS. Trace element levels were then compared with water from two other rivers (Maha Oya, Kalu Ganga) and from six dry zone irrigation reservoirs. Results showed that the trace metal concentrations in the Mahaweli upper catchment were detected in the order of Fe > Cu > Zn > Se > Cr > Mn > As > Ni > Co > Mo. Remarkably high levels of Ca, Cr, Co, Ni, Cu, As, and Se were observed in the Mahaweli Basin compared to other study rivers. Considerably high levels of Cr, Mn, Fe, Co, Ni, Cu, Zn, As, and Se were found in upstream tributaries of the Mahaweli River. Such metals possibly originated from phosphate and organic fertilizers that are heavily applied for tea and vegetable cultivations within the drainage basin. Cadmium that is often attributed to the etiology of unknown chronic kidney diseases in certain parts of the dry zone is much lower than previously reported levels. Decrease in these metals in the lower part of the Mahaweli River could be due to adsorption of trace metals onto sediment and consequent deposition in reservoirs.

Impact of wastewater from different sources on the prevalence of antimicrobial-resistant Escherichia coli in sewage treatment plants in South India.

The sewage treatment plant (STP) is one of the most important interfaces between the human population and the aquatic environment, leading to contamination of the latter by antimicrobial-resistant bacteria. To identify factors affecting the prevalence of antimicrobial-resistant bacteria, water samples were collected from three different STPs in South India. STP1 exclusively treats sewage generated by a domestic population. STP2 predominantly treats sewage generated by a domestic population with a mix of hospital effluent. STP3 treats effluents generated exclusively by a hospital. The water samples were collected between three intermediate treatment steps including equalization, aeration, and clarification, in addition to the outlet to assess the removal rates of bacteria as the effluent passed through the treatment plant. The samples were collected in three different seasons to study the effect of seasonal variation. Escherichia coli isolated from the water samples were tested for susceptibility to 12 antimicrobials. The results of logistic regression analysis suggest that the hospital wastewater inflow significantly increased the prevalence of antimicrobial-resistant E. coli, whereas the treatment processes and sampling seasons did not affect the prevalence of these isolates. A bias in the genotype distribution of E. coli was observed among the isolates obtained from STP3. In conclusion, hospital wastewaters should be carefully treated to prevent the contamination of Indian environment with antimicrobial-resistant bacteria.

Hazardous implications of halogenated polycyclic aromatic hydrocarbons in feedstuff: Congener specificity and toxic levels in feed ingredients and feeds.

Studies have shown that halogenated polycyclic aromatic hydrocarbons (HPAHs), including chlorinated (ClPAHs) and brominated PAHs (BrPAHs), could be hazardous pollutants due to their pervasive occurrence in the environment. However, their accumulation properties and toxic potentials in animal feedstuffs remain unclear. This study investigated 75 congeners of parent PAHs, ClPAHs, and BrPAHs in animal-based feed ingredients and fish and swine feeds in Japan using a GC/Orbitrap MS system. The total parent PAHs ranged from below the method detection limit (

Plastiome: Plastisphere-enriched mobile resistome in aquatic environments.

Aquatic microplastics (MPs) act as reservoirs for microbial communities, fostering the formation of a mobile resistome encompassing diverse antibiotic (ARGs) and biocide/metal resistance genes (BMRGs), and mobile genetic elements (MGEs). This collective genetic repertoire, referred to as the "plastiome," can potentially perpetuate environmental antimicrobial resistance (AMR). Our study examining two Japanese rivers near Tokyo revealed that waterborne MPs are primarily composed of polyethylene and polypropylene fibers and sheets of diverse origin. Clinically important genera like Exiguobacterium and Eubacterium were notably enriched on MPs. Metagenomic analysis uncovered a 3.46-fold higher enrichment of ARGs on MPs than those in water, with multidrug resistance genes (MDRGs) and BMRGs prevailing, particularly within MPs. Specific ARG and BMRG subtypes linked to resistance to vancomycin, beta-lactams, biocides, arsenic, and mercury showed selective enrichment on MPs. Network analysis revealed intense associations between host genera with ARGs, BMRGs, and MGEs on MPs, emphasizing their role in coselection. In contrast, river water exhibited weaker associations. This study underscores the complex interactions shaping the mobile plastiome in aquatic environments and emphasizes the global imperative for research to comprehend and effectively control AMR within the One Health framework.

Profiles of perfluoroalkyl substances in the liver and serum of patients with liver cancer and cirrhosis in Australia.

The present cross-sectional study investigated 12 perfluoroalkyl substances (PFASs) in serum (n=79) and liver (n=66) samples from patients who had undergone liver transplantation for a range of conditions, such as hepatocellular carcinoma (HCC), cirrhosis due to chronic hepatitis C viral infection (HCV), both HCC and HCV, amyloidosis or acute liver failure. PFAS data from patients were compared to those in control serum (n=25) samples from liver donors with no known liver disease and to those in control liver (n=9) tissues collected during liver resection surgery. All samples showed detectable PFOS (serum: 0.621-126ng/mL; liver: 0.375-42.5ng/g wet wt) and PFOA (serum: 0.437-45.5ng/mL; liver: 0.101-2.25ng/g wet wt) concentrations. In general, in paired serum and liver samples, serum had higher PFOS, PFHxS, PFDA, PFNA, and PFOA concentrations than those in explanted livers from patients. These findings also suggest that pathological changes in diseased livers alter the distribution of PFASs between liver and serum. The results from control serum (2007-2008) suggested that PFOS, PFHxS, PFOA, and PFNA concentrations were lower than those previously reported from Australia for 2002-2003, and 2006-2007. The present study demonstrates, for the first time, the detection and comparison of a range of PFASs in the liver of patients with liver cancer and/or liver cirrhosis.

Perfluoroalkyl substances in the blood of wild rats and mice from 47 prefectures in Japan: use of samples from nationwide specimen bank.

Numerous studies have reported on the global distribution, persistence, fate, and toxicity of perfluoroalkyl and polyfluoroalkyl substances (PFASs). However, studies on PFASs in terrestrial mammals are scarce. Rats can be good sentinels of human exposure to toxicants because of their habitat, which is in close proximity to humans. Furthermore, exposure data measured for rats can be directly applied for risk assessment because many toxicological studies use rodent models. In this study, a nationwide survey of PFASs in the blood of wild rats as well as surface water samples collected from rats' habitats from 47 prefectures in Japan was conducted. In addition to known PFASs, combustion ion chromatography technique was used for analysis of total fluorine concentrations in the blood of rats. In total, 216 blood samples representing three species of wild rats (house rat, Norway rats, and field mice) were analyzed for 23 PFASs. Perfluorooctanesulfonate (PFOS; concentration range <0.05-148 ng/mL), perfluorooctane sulfonamide (PFOSA; <0.1-157), perfluorododecanoate (<0.05-5.8), perfluoroundecanoate (PFUnDA; <0.05-51), perfluorodecanoate (PFDA; <0.05-9.7), perfluorononanoate (PFNA; <0.05-249), and perfluorooctanoate (PFOA) (<0.05-60) were detected >80 % of the blood samples. Concentrations of several PFASs in rat blood were similar to those reported for humans. PFSAs (mainly PFOS) accounted for 45 % of total PFASs, whereas perfluoroalkyl carboxylates (PFCAs), especially PFUnDA and PFNA, accounted for 20 and 10 % of total PFASs, respectively. In water samples, PFCAs were the predominant compounds with PFOA and PFNA found in >90 % of the samples. There were strong correlations (p < 0.001 to p < 0.05) between human population density and levels of PFOS, PFNA, PFOA, and PFOSA in wild rat blood.

Occurrence and the potential ecological risk of veterinary antimicrobials in swine farm wastewaters in Japan: Seasonal changes, relation to purchased quantity and after termination of oxytetracycline usage.

This study provides the first comprehensive investigation of the residual concentrations of eight classes of antimicrobial agents (AMs, 20 compounds) in 13 swine wastewater treatment facilities in Japan. These facilities implemented the aerobic activated sludge (AS) or its alternative methods. The maximum concentrations before treatment were found at the level of 7100, 6900, 6000, 3600, 3400, and 1400 µg/L for tilmicosin, oxytetracycline (OTC), chlortetracycline, lincomycin , sulfamethoxazole, and trimethoprim, respectively. The highest detection rate (96.3%) in influents was noted for the morantel, which was a feed additive. The seasonal difference in residual concentration was much greater for tetracyclines (TCs) and macrolides (MLs) when their residual concentrations were high, especially in the cold season. There was a positive correlation between the purchased quantity of TCs and fluoroquinolones (FQs) and their residue levels detected in the effluents (p < 0.01). The estimated removal rate of AMs was greater than 80%. In contrast, on a few occasions, it was diminished due to failing operating conditions, such as water temperature and AS rate in the aeration tank. The estimated ecological risks of AMs in effluents based on risk quotients (RQs) considered to enhance the selection pressure for drug resistance (RQs-AMR) were high for TCs and FQs, whereas ecotoxicological effects (RQs-ENV) to aquatic organisms were higher for sulfonamides and MLs. When OTC usage ceased, its concentration in wastewater decreased rapidly; however, it remained longer period in the effluents, probably due to OTC desorption from the AS. The concentrations (and respective RQs) of TCs were decreased by >99.8% and >92% in the influents and effluents, respectively. This data suggested that it is essential to reduce the amount used and introduce more efficient methods and operating conditions to constantly remove AMs during the treatment to reduce the risk of AM discharge from swine farms.

Characteristics of antimicrobial residues in manure composts from swine farms: Residual patterns, removal efficiencies, and relation to purchased quantities and composting methods in Japan.

Present study provides first comprehensive results on the residual levels of 19 antimicrobial (AM) residues in 12 Japanese swine manure composting facilities that use open or enclosed types of treatment methods. Tilmicosin (14000 µg/kg d.w.) and tiamulin (15000 µg/kg d.w.) were present in the highest concentrations in manure composts. Morantel (MRT) had the highest detection frequency (100%) in compost, suggesting its ubiquitous usage and resistance to degradation during composting. Sulfamethoxazole had low detection frequencies and concentrations, likely due to limited partitioning to the solid phase. A positive correlation (p < 0.05) between purchasing quantities and residue levels in manure composts was detected for fluoroquinolones (FQs). The removal efficiencies of AMs in enclosed-type facilities were lower and more inconsistent than those in open-type facilities. Tetracyclines (TCs), lincomycin, and trimethoprim were easily removed from open-type facilities, whereas FQs and MRT persisted in both facilities. After discontinuing the usage of oxytetracycline (OTC), TCs concentrations reduced drastically in input materials, remained pseudo-persistent in composts for up to 4 months, suggesting a time lag for composting and were not detected (<10 µg/kg) after 4 months of OTC withdrawal. This study emphasizes on the effectiveness of manure composting methods in reducing AM residues in swine waste.

Voyaging of halogenated polycyclic aromatic hydrocarbons, an emerging group of pollutants, on micro-mesoplastics in the marine environment.

The limited existing research on the accumulation of hazardous chlorinated and brominated polycyclic aromatic hydrocarbons (ClPAHs and BrPAHs) in micro-mesoplastics (mMPs) motivated this investigation. We collected mMPs from the coastal environments of Sri Lanka and Japan. Out of 75 target compounds analyzed, 61 were detected, with total parent PAH concentrations reaching 16,300 and 1770 ng/g plastic in Sri Lanka and Japan, respectively. The total parent PAH concentrations in mMPs from the southern Sri Lankan coastline were relatively higher than those from the eastern coastline. Phenanthrene and naphthalene were the dominant parent PAH congeners in most mMP samples. Chlorinated pyrenes and brominated naphthalene were predominant among halogenated PAHs. The estimated toxic equivalency quotient (TEQ) ranged from 0.67 to 1057 ng-TEQ/g plastic, with the highest levels observed in polystyrene (PS) particles from the southern Sri Lankan coast. Benzo[a]pyrene and dibenzo[a,h]anthracene exhibited elevated TEQ for parent PAHs, whereas dichloropyrene, and dibromopyrene represented the highest TEQs for ClPAHs and BrPAHs, respectively. The data evidenced that several HPAH congeners can increase the PAH-like toxicity (∼86%) in mMPs. This study provides insights into the accumulation of parent and halogenated PAHs in mMPs, highlighting their potential combined implications in marine and terrestrial ecosystems.

Presence of Halogenated Polycyclic Aromatic Hydrocarbons in Milk Powder and the Consequence to Human Health.

Recent reports of the presence of halogenated derivatives of polycyclic aromatic hydrocarbons (PAHs) in human foods of animal origin, such as chlorinated (ClPAHs) and brominated (BrPAHs) PAHs, suggest that their contamination in dairy products may also pose a human health risk. This study used GC/Orbitrap-MS to analyze 75 congeners of halogenated PAHs and parent PAHs in milk and creaming powder samples commonly found in grocery stores in Sri Lanka and Japan. Our investigation revealed a total of 31 halogenated PAHs (HPAHs) in the samples. The concentrations of total parent PAHs in the samples from Sri Lanka and Japan ranged from not detected (n.d.)−0.13 and <0.001−16 ng/g dry weight (d.w.). Total ClPAHs and BrPAHs in the samples ranged from 0.01−3.35 and 1.20−5.15 ng/g (d.w.) for Sri Lanka, and 0.04−2.54 and n.d.−2.03 ng/g d.w. for Japan, respectively. The ClPAHs were dominated by chlorinated-pyrene, -fluoranthene, and -benzo[a]pyrene congeners, whereas the BrPAHs were dominated by brominated-naphthalene and -pyrene congeners. The toxic assessment estimated based on the intake of toxic equivalency quotients (TEQs) for target compounds in milk powders revealed that HPAHs might contribute additively to the PAHs-associated health risk to humans, indicating that more research is needed.

Occurrence of Pharmaceutically Active Compounds and Potential Ecological Risks in Wastewater from Hospitals and Receiving Waters in Sri Lanka.

The presence of pharmaceutically active compounds (PACs) in the environment and their associated hazards is a major global health concern; however, data on these compounds are scarce in developing nations. In the present study, the existence of 39 non-antimicrobial PACs and six of their metabolites in wastewater from hospitals and adjacent surface waters in Sri Lanka was investigated from 2016 to 2018. The highest amounts of the measured chemicals, including the highest concentrations of atorvastatin (14,620 ng/L) and two metabolites, mefenamic acid (12,120 ng/L) and o-desmethyl tramadol (8700 ng/L), were detected in wastewater from the largest facility. Mefenamic acid, gemfibrozil, losartan, cetirizine, carbamazepine, and phenytoin were detected in all the samples. The removal rates in wastewater treatment were 100% for zolpidem, norsertaline, quetiapine, chlorpromazine, and alprazolam. There was substantial variation in removal rates of PACs among facilities, and the overall data suggest that treatment processes in facilities were ineffective and that some PAC concentrations in the effluents were increased. The estimated risk quotients revealed that 14 PACs detected in water samples could pose low to high ecological risk to various aquatic organisms. Compounds such as ibuprofen, tramadol, and chlorpromazine detected in untreated and treated wastewater at these facilities pose a high risk to several aquatic organisms. Our study provides novel monitoring data for non-antimicrobial PAC abundance and the associated potential ecological risk related to hospitals and urban surface waters in Sri Lanka and further offers valuable information on pre-COVID-19 era PAC distribution in the country. Environ Toxicol Chem 2022;41:298-311. © 2021 SETAC.

Comparative Study on the Distribution of Essential, Non-Essential Toxic, and Other Elements across Trophic Levels in Various Edible Aquatic Organisms in Sri Lanka and Dietary Human Risk Assessment.

Thirty-six elements are categorized as essential but toxic in excess amount (EBTEs), non-essential toxic (NETs), and Other in 29 different edible aquatic species dwelling in offshore pelagic, and coastal and estuarine (CE) ecosystems were investigated in Sri Lanka. Elements were analyzed using an energy-dispersive X-ray fluorescence (EDXRF) spectrometer, and an NIC MA-3000 Mercury Analyzer. EBTEs showed a negative relationship, whereas NETs showed a positive relationship between the concentration (mg/kg wet weight) and trophic levels in both ecosystems. EBTEs showed trophic dilution, whereas NETs showed trophic magnification. Some elements in a few organisms exceeded the maximum allowable limit which is safe for human consumption. There was a positive relationship (R2 = 0.85) between the concentration of mercury and body weight of yellowfin tuna (YFT). For the widely consumed YFT, the calculated hazard index (HI) for the non-carcinogenic health and exposure daily intake of NETs for adults were 0.27 and 9.38 × 10-5 mg/kg bw/day, respectively. The estimated provisional tolerable weekly intake (PTWI) (µg/kg bw/w) was 0.47 for arsenic and 0.05 for antimony, cadmium, mercury, and lead. The HI and PTWI values were below the recommended limits; thus, consumption of YFT does not pose any health risk for Sri Lankan adults.

Occurrence and exposure assessment of perchlorate, iodide and nitrate ions from dairy milk and water in Japan and Sri Lanka.

Perchlorate is known to competitively interfere with iodide uptake by the thyroid gland and thereby human exposure to perchlorate is a public health concern. Prevalence of perchlorate in dairy milk is documented; nevertheless, co-occurrence of perchlorate with other thyroid-binding monovalent ions such as iodide and nitrate is not well understood. In this study, we analyzed perchlorate, iodide, and nitrate-N in dairy milk, water and other dairy-related samples collected from Japan and Sri Lanka. Concentrations of perchlorate in Japanese dairy milk samples ranged from 1.03 to 14.1 ng ml(-1); the corresponding concentrations in dairy milk and powdered milk from Sri Lanka were 1.14-38.5 ng ml(-1). Perchlorate concentrations in commercial milk were significantly higher in Japan than in Sri Lanka, while iodide and nitrate levels in milk between the two countries were comparable. All three ions were ubiquitously found in water samples from Japan and Sri Lanka. Analysis of colostrum and raw milk collected from cows fed with the same feed for over 30 days showed no significant temporal variations in perchlorate, iodide and nitrate-N concentrations. A significant positive correlation was found between the concentrations of perchlorate and iodide in Japanese commercial milk. The concentrations of perchlorate and nitrate-N in water samples analyzed from both countries also showed a significant positive correlation. The exposure estimation revealed that dairy milk provides a greater source for perchlorate and iodide, while water predominantly contributes nitrate-N intake for all age groups in both counties. Infants and children demonstrated the highest estimated perchlorate, iodide and nitrate-N intake on a body weight basis in comparison to other age groups. Therefore, further studies of risk associated with perchlorate may need to reconsider co-existence of iodine and other iodide transport inhibitors in food.

Halogenated polycyclic aromatic hydrocarbons in edible aquatic species of two Asian countries: Congener profiles, biomagnification, and human risk assessment.

Seventy-five contaminants including chlorinated/brominated/parent polycyclic aromatic hydrocarbons (Cl/Br/PAHs) were investigated in 29 edible aquatic species from the Indian Ocean near Sri Lanka and 10 species from the Pacific Ocean near Japan. Concentrations of total ClPAHs and BrPAHs in the samples were 2.6-57 and 0.30-9.5 ng/g-dry weight from the Indian Ocean, and 0.35-18 and 0.03-3.3 ng/g-dry weight from the Pacific Ocean, respectively. Comparing the profiles of Cl/BrPAHs among the samples, congeners of chlorinated and brominated pyrene were predominant components and enhanced the potential for biomagnification in the sample from the off-shore pelagic environment in the Indian Ocean. The incremental lifetime cancer risks estimated by intake of the targets in consuming aquatic organisms showed that approximately one-third of studied organisms exceeded the acceptable risk level for Sri Lankans.

The association between antimicrobials and the antimicrobial-resistant phenotypes and resistance genes of Escherichia coli isolated from hospital wastewaters and adjacent surface waters in Sri Lanka.

The presence of antimicrobials, antimicrobial-resistant bacteria (ARB), and the associated antimicrobial resistance genes (ARGs) in the environment is a global health concern. In this study, the concentrations of 25 antimicrobials, the resistance of Escherichia coli (E. coli) strains in response to the selection pressure imposed by 15 antimicrobials, and enrichment of 20 ARGs in E. coli isolated from hospital wastewaters and surface waters were investigated from 2016 to 2018. In hospital wastewaters, clarithromycin was detected at the highest concentration followed by sulfamethoxazole and sulfapyridine. Approximately 80% of the E. coli isolates were resistant, while 14% of the isolates exhibited intermediate resistance against the tested antimicrobial agents. Approximately 61% of the examined isolates were categorized as multidrug-resistant bacteria. The overall abundance of phenotypes that were resistant toward drugs was in the following order: ß-lactams, tetracycline, quinolones, sulfamethoxazole/trimethoprim, aminoglycosides, and chloramphenicol. The data showed that the E. coli isolates frequently harbored blaTEM, blaCTX-M, tetA, qnrS, and sul2. These results indicated that personal care products were significantly associated with the presence of several resistant phenotypes and resistance genes, implying their role in co-association with multidrug resistance. Statistical analysis also indicated a disparity specific to the site, treatment, and year in the data describing the prevalence of ARB and ARGs and their release into downstream waters. This study provides novel insights into the abundance of antimicrobial, ARB and ARGs in Sri Lanka, and could further offer invaluable information that can be integrated into global antimicrobial resistance databases.

Synergistic induction of drug-metabolizing enzymes in co-cultures of bovine hepatocytic and sinusoidal cell lines.

Hepatocyte-derived cell lines provide useful experimental systems for studying liver metabolism. Unlike human and rodents, few hepatocyte-derived cell lines have been generated from cattle. Here, we established two immortalized bovine hepatocyte-derived cell lines (BH4 and BH5) via transfection with a SV40 large T-antigen construct. Morphological and immunocytochemical analyses revealed that BH4 and BH5 originated from hepatocytes and biliary-epithelial cells, respectively. A potent carcinogen, 3-methylcholanthrene (3-MC), upregulated gene expression of cytochrome P450 (CYP)1A1, CYP1A2, and CYP1B1 in BH4 and BH5, but only to levels less than one-fifteenth of those in primary cultured bovine hepatocytes. Phenobarbital (PB) also increased expression levels of CYP2B6, CYP2C18, and CYP3A4 in BH4 and BH, but at a lower level than 3-MC. By contrast, when BH4 or BH5 was co-cultured with previously established bovine liver sinusoidal cell lines and treated with 3-MC, the gene expression levels of CYP1A1, CYP1A2, and CYP1B1 increased by 38~290%, compared with those in BH4 or BH5 cells cultured alone. PB-treated co-cultures of BH4 or BH5 cells and liver sinusoidal cell lines also showed synergistic increases in CYP2B6 and CYP2C18 expression. Together, the results suggest that these co-cultures could provide an in vitro model for investigations into pharmacological and toxicological properties of drugs in cattle liver.

Accumulation of halogenated polycyclic aromatic hydrocarbons by different tuna species, determined by high-resolution gas chromatography Orbitrap mass spectrometry.

Halogenated polycyclic aromatic hydrocarbon (HPAH) concentrations in tissues from three tuna species Thunnus albacares (yellowfin tuna), Katsuwonus pelamis (skipjack tuna), and Auxis thazard (frigate tuna) were determined by high-resolution gas chromatography Orbitrap mass spectrometry. The tuna samples were collected from the Indian Ocean. The instrument conditions gave high mass accuracy at 0.9 m/z isolation width of the mass filter and a mass error of <±1.0 ppm for many HPAHs. A total of 29 of the 30 targets chlorinated PAHs (ClPAHs) and 20 of the 21 targets brominated PAHs (BrPAHs) were detected in the tuna muscle samples. The mean total ClPAH, BrPAH and PAH concentrations for tuna were 127.2, 156.6 and 682.8 ng/g lipid weight, respectively. The mean total ClPAH and BrPAH concentrations (ng/g lipid weight) in the tuna were considerably lower than that of PAH concentrations. The mean total ClPAH, BrPAH and PAH concentrations in T. albacares respectively were 185.8, 249.2 and 784.1 ng/g lipid weight, irrespective of the body sizes. The mean total ClPAH, BrPAH and PAH concentrations in K. pelamis respectively were 45.1, 24.8 and 555.6 ng/g lipid weight. The mean total ClPAH, BrPAH and PAH concentrations in A. thazard respectively were 34.09, 4.73 and 433.24 ng/g lipid weight. The total ClPAH concentrations and body weights significantly positively correlated for T. albacares. The mean total ClPAH concentration in white muscles was significantly higher (p < 0.05) for large than for small T. albacares. This suggests ClPAHs could bioaccumulate in T. albacares, possibly because they are poorly metabolized. The chlorinated phenanthrene and pyrene concentrations indicated tuna accumulate these compounds increasingly effectively as the tuna grow. This was the first time large numbers of HPAHs were found in biological samples. HPAHs may adversely affect the health of humans consuming tuna.