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Studies on the conditions and pollution routes of antibiotic-resistant bacteria (ARB) in rivers can help provide countermeasures against the spread of ARB. This study focused on the pristine uppermost stream of a river, where Escherichia coli (E. coli) and enterococci were detected, although the stream flows through a pristine forest catchment. Antibiotic resistance of E. coli and enterococci isolated from the river water, riverbed sediment, and feces of waterside animals, such as birds and Mustelidae, were investigated throughout the year in the pristine uppermost sites. Antibiotic resistance was present in 1.4% (7/494) of the E. coli strains and 3.0% (24/812) of the enterococcal strains, and was low throughout the year. Although antibiotic resistance of bacteria isolated from feces was not detected in this watershed, the prevalence of multidrug-resistant E. coli was 0.4% (1/246) and 0.6% (1/172) in river water and riverbed sediment samples, respectively were observed. The presence of extended-spectrum β-lactamase (ESBL)-producing E. coli was confirmed in river water samples, and genomic analysis revealed that the samples possessed the CTX-M-15 group. Multidrug-resistant strains and ESBL-producing strains were classified as phylogroups B1 and A, respectively, which are E. coli phenotypes isolated from wild animals. Pulsed-field gel electrophoresis revealed analysis targeting enterococci that strains isolated from river water and bird feces were in the same cluster with 100% similarity. Therefore, bird feces are a source of enterococci in the uppermost stream of the river. Because multidrug-resistant bacteria and ESBL-producing bacteria were present in the pristine uppermost stream of the pristine river, urgent elucidation of the spreading routes of ARB is important.

In the ballasted flocculation, high-speed sedimentation of suspensions is achieved using a microsand as a ballast material and a polymer flocculant combined with microflocs made of polyaluminum chloride (PAC) as an inorganic coagulant. In this study, three turbid water samples containing kaolin clay (kaolin concentration: 20, 200, and 500 mg/L) were treated by coagulation–sedimentation and ballasted flocculation. The effects of pH and PAC dosage, which are the controlling parameters for coagulation, and the microsand (silica sand) and polymer dosages, which are the controlling parameters for ballasted treatment, on the treatment efficiency and floc settling velocity were examined. The floc settling velocity under the optimum conditions was 17 times higher than that of the conventional coagulation–sedimentation process using PAC. The turbidity was 0.54 turbidity unit (TU) (TU as the kaolin standard), and its removal efficiency was 99.7%. Furthermore, turbid water samples with different kaolin concentrations (20 and 500 mg/L) were treated via the ballasted flocculation. In this study, fundamental information on the optimization of each dosage condition of coagulant, ballast, and polymer and pH condition in ballasted flocculation was obtained, and the removal mechanisms under optimal, underoptimistic and overoptimistic conditions were proposed. HIGHLIGHTS Conditions for ultrahigh-speed ballast flocculation were investigated.; Floc settling velocity increased and showed a peak at 10 g/L of microsand.; Excess addition of microsand significantly reduced the floc settling velocity.; Hugh floc settling velocity was achieved in the treatment of high-turbidity water.;

Enterococcus spp. are common bacteria present in the intestinal tracts of animals and are used as fecal indicators in aquatic environments. On the other hand, enterococci are also known as opportunistic pathogens. Elucidating their composition in the intestinal tracts of domestic animals can assist in estimating the sources of fecal contamination in aquatic environments. However, information on the species and composition of enterococci in animal hosts (except humans) is still lacking. In this study, enterococci were isolated from the feces of cattle, pigs, birds, and humans using selective media. Enterococcal species were identified using mass spectrometry technology, and each host was characterized by diversity and cluster analysis. The most dominant species were E. hirae in cattle, E. faecium in birds, and E. faecalis in pigs and humans. Cattle had the highest alpha diversity, with high interindividual and livestock farm diversity. The dominant enterococcal species in pigs and humans were identical, and cluster analysis showed that the majority of the two hosts’ species clustered together.

High counts of bacteria are present in beach sand, and human health threats attributable to contact with sand have been reported. In this study, we investigated fecal indicator bacteria in the top surface sand of coastal beaches. Monitoring investigations were performed during a monsoon when rainfall occurs randomly, and the composition of the coliforms was analyzed. The coliform count in the top surface sand (depth < 1 cm) increased by approximately 100 fold (26–2.23 × 103 CFU/100 g) with increasing water content because of precipitation. The composition of the coliforms in the top surface sand changed within 24 h of rainfall, with Enterobacter comprising more than 40% of the coliforms. Estimation of factors that changed the bacterial counts and composition revealed that coliform counts tended to increase with increasing water content in the top surface sand. However, the abundance of Enterobacter was independent of the sand surface temperature and water content. Coliform counts in the top surface sand rapidly increased and the composition showed remarkable variations because of the supply of water to the beach following rainfall. Among them, some bacteria with suspected pathogenicity were present. Controlling bacteria in coastal beaches is important for improving public health for beachgoers.

One of the major challenges in detecting waterborne pathogens is the low concentration of the target bacteria in water. In this study, we applied the coagulation and foam concentration method to obtain DNA from water samples collected from upstream, near an estuary. The DNA samples were analyzed using 16S rRNA gene sequencing to clarify the microbial community shifts and to identify potentially pathogenic bacteria. Bacterial communities changed as the river flowed downstream, most likely influenced by land use and human activities such as the discharge of wastewater-treatment plant effluent. Based on the 16S rRNA gene amplicon sequencing, potentially pathogenic bacteria were detected with greater than 0.1% of their relative abundances. Among these, Yersinia ruckeri and Pseudomonas alcaligenes were widely detected in the river water. In addition, digital PCR (dPCR) was used to quantify major waterborne pathogens. Shiga toxin-producing Escherichia coli (STEC), Shigella spp., and Campylobacter jejuni were all below the limit of detection. In contrast, general E. coli, which has the beta-D-glucuronidase gene (uidA) were detected by dPCR (copies/100 mL) at similar levels to those measured using the culture-based method (as colony forming units/100 mL). These results suggest that the coagulation and foam concentration method is useful for concentrating microbes and obtaining DNA from river water samples for environmental monitoring.