Sewage input and the relationship between chemical markers (linear alkylbenzenes and coprostanol) and fecal indicator bacteria (FIB, Escherichia coli and enterococci), were evaluated in order to establish thresholds values for chemical markers in suspended particulate matter (SPM) as indicators of sewage contamination in two subtropical estuaries in South Atlantic Brazil. Both chemical markers presented no linear relationship with FIB due to high spatial microbiological variability, however, microbiological water quality was related to coprostanol values when analyzed by logistic regression, indicating that linear models may not be the best representation of the relationship between both classes of indicators. Logistic regression was performed with all data and separately for two sampling seasons, using 800 and 100 MPN 100 mL⁻¹ of E. coli and enterococci, respectively, as the microbiological limits of sewage contamination. Threshold values of coprostanol varied depending on the FIB and season, ranging between 1.00 and 2.23 μg g⁻¹ SPM. The range of threshold values of coprostanol for SPM are relatively higher and more variable than those suggested in literature for sediments (0.10–0.50 μg g⁻¹), probably due to higher concentration of coprostanol in SPM than in sediment. Temperature may affect the relationship between microbiological indicators and coprostanol, since the threshold value of coprostanol found here was similar to tropical areas, but lower than those found during winter in temperate areas, reinforcing the idea that threshold values should be calibrated for different climatic conditions.

In respect to direct and indirect water reuse, the microbiological quality of treated wastewater is highly important. Conventional wastewater treatment plants are normally not equipped with advanced technologies for the elimination of bacteria. Molecular biology analyses were combined with live-dead discrimination analysis of wastewater population using Propidium monoazide (PMA) to study population shifts during ozonation (1 g ozone/g DOC) at a municipal wastewater treatment plant. Escherichia coli, enterococci, and Pseudomonas aeruginosa were quantified by polymerase chain reaction (qPCR) and the whole wastewater population was analyzed by metagenomic sequencing. The PMA-qPCR experiments showed that the abundances of P. aeruginosa didn't change by ozone treatment, whereas a reduction was observed for E. coli and enterococci. Results comparing conventional cultivation experiments with PMA-qPCR underlined the presence of viable but not culturable cells (VBNC) and their regrowth potential after ozone treatment. Illumina HiSeq sequencing results with and without PMA treatment demonstrated high population similarities in water samples originating from ozone inflow sampling sides. Upon using PMA treatment after ozonation, population shifts became visible and also underlined the importance of PMA treatment for the evaluation of elimination and selection processes during ozonation at WWTPs. Amongst a number of 14 most abundant genera identified in the inflow samples, 9 genera were found to be reduced, whereas 4 genera increased in relative abundance and 1 genus almost remained constant. The strongest increase in relative abundance after ozonation was detected for Oscillatoria spp., Microcoleus spp. and Nitrospira spp. Beside this, a continuous release of Pseudomonas spp. (including P. aeruginosa) to the downstream receiving body was confirmed. Regrowth experiments demonstrated a high prevalence of P. aeruginosa as part of the surviving bacterial population. Summing up, molecular biology analyses in combination with live-dead discrimination are comprehensive methods to evaluate the elimination processes targeting specific species and/or whole microbial populations.

Ballast water is used to safely stabilize and operate shipping vessels worldwide, in a multitude of aquatic settings, including inland, coastal and open oceans. However, ballast water may pose ecological, public health, and/or economic problems as it may serve as vehicles of transmission of microorganisms. Current ballast water regulations include limits of Escherichia coli, Enterococcus spp. and toxigenic Vibrio cholerae. Several United States Environmental Protection Agency approved standard operating protocols (SOPs) exist for detection of E. coli and Enterococci, yet none exists for V. cholerae. Current V. cholerae detection methods include colony blot hybridization, direct fluorescent antibody test (DFA), and/or polymerase chain reaction (PCR), which can be time consuming and difficult to perform. This study utilizes Cholera SMART II to determine its potential use in detection of V. cholerae. Validation of this method would help provide quick and accurate analysis for V. cholerae in ballast discharge waters in the field.

Hong Kong's beach water quality classification scheme, used effectively for >25 years in protecting public health, was first established in local epidemiology studies during the late 1980s where Escherichia coli (E. coli) was identified as the most suitable faecal indicator bacteria. To review and further substantiate the scheme's robustness, a performance check was carried out to classify water quality of 37 major local beaches in Hong Kong during four bathing seasons (March–October) from 2010 to 2013. Given the enterococci and E. coli data collected, beach classification by the local scheme was found to be in line with the prominent international benchmarks recommended by the World Health Organization and the European Union. Local bacteriological studies over the last 15 years further confirmed that E. coli is the more suitable faecal indicator bacteria than enterococci in the local context.

Bathing water quality plays a key role for public health, is highly important for recreational tourism and therefore monitored in the EU-Directive 2006/7/EC. To identify pollution hot spots, sources and impacts of the directive-change in 2006, including a change of indicator organisms, we evaluated monitoring data of the past 15 years, collected own data, determined survival rates of indicator organisms and applied hydrodynamic modelling in a micro-tidal-system.Due to higher survival rates under turbid conditions and restricted water exchange, shallow, eutrophic bays and lagoons are hot spots of microbial pollution. Rain events cause high microbial emission and distribution. Based on different decay rates, the ratio of E. coli to Enterococci can hint towards a pollution source. Including rain predictions, currents and winds, hydrodynamic models can then assess the daily risk of microbial pollution at each bathing site. They are an important tool to modify beach management and event-based monitoring.

Water quality in Lake Pontchartrain was deteriorating and recreational activities along the beach were restricted by the end of the 20th Century. A microbial source tracking (MST) study was conducted to determine the fecal contamination sources at public beach of the lake, so that effective pollution control strategies can be developed. Water samples were collected over an eight-month period at ten locations along the lake in 2016 and 2017. E. coli and Enterococcus were detected in 90.6% (culture) and 97.5% (qPCR), 95.8% (culture) and 91.8% (qPCR) of water samples from all sampling sites, respectively. Significant positive relationship between E. coli and Enterococcus results was observed for both qPCR and culture methods. HF183 marker was detected in 94.3% water samples (149 of 158), with concentrations ranging from 29.0 to 6073.5GC/100ml and from 129.8 to 38,465.6GC/100ml in summer and winter, respectively. The results also indicate that significant rainfall events have the potential to supply considerable loads of fecal bacteria to lake waters. Further research is needed to determine the contribution of other animals to fecal contamination in the region.

The research goal was to determine if onsite wastewater system (OWS) density had an influence on the concentrations and watershed exports of Escherichia coli and enterococci in urbanizing watersheds. Eight watersheds with OWS densities ranging from < 0.1 to 1.88 systems ha⁻¹ plus a watershed served by sewer (Sewer) and a mostly forested, natural watershed (Natural) in the Piedmont of North Carolina served as the study locations. Stream samples were collected approximately monthly during baseflow conditions between January 2015 and December 2016 (n = 21). Median concentrations of E. coli (2014 most probable number (MPN) 100 mL⁻¹) and enterococci (168 MPN 100 mL⁻¹) were elevated in streams draining watersheds with a high density of OWS (> 0.77 system ha⁻¹) relative to watersheds with a low (< 0.77 system ha⁻¹) density (E. coli: 204 MPN 100 mL⁻¹ and enterococci: 88 MPN 100 mL⁻¹) and control watersheds (Natural: E. coli: 355 MPN 100 mL⁻¹ and enterococci: 62 MPN 100 mL⁻¹; Sewer: 177 MPN 100 mL⁻¹ and 130 MPN 100 mL⁻¹). Samples collected from watersheds with a high density of OWS had E. coli and enterococci concentrations that exceeded recommended thresholds 88 and 57% of times sampled, respectively. Results show that stream E. coli and enterococci concentrations and exports are influenced by the density of OWS in urbanizing watersheds. Cost share programs to help finance OWS repairs and maintenance are suggested to help improve water quality in watersheds with OWS.

The relationship between land use and microbial community structure at seven sites along the Lower Mekong River, between Thailand and the Loa People’s Democratic Republic, was investigated using Illumina next-generation sequencing of the V5–V6 hypervariable regions of the 16S rRNA gene. In total, 14,470 operational taxonomic units (OTUs) were observed. Community composition was significantly different between sampling years. Moraxellaceae and Comamonadaceae were the predominant bacterial families in upstream sites, which included agricultural and urban areas in the Loei and Nong Khai provinces of Thailand. Members of the family Comamonadaceae were prevalent in agricultural and urban sites in Bueng Kan Province, while Moraxellaceae and Burkholderiaceae were the major families in a site downstream of an urban area in the Nakhon Phanom Province of Thailand. The bacterial community observed from a forested area of Patam National Park in Thailand showed greatest diversity, and several major bacterial families including Comamonadaceae, Moraxellaceae, and Pseudomonadaceae were more dominant than other sites. The diversity of fecal indicator bacteria, determined by ERIC-PCR DNA fingerprinting, indicated the presence of 29 strains of Escherichia coli and 21 strains of Enterococcus, while TP-RAPD patterns represented six species of Enterococcus. Results of this study indicated that although the difference in the distribution of bacterial phyla and families was found among sampling sites, the bacterial community composition, based on the presence of OTUs, continuously retained its signature across approximately 758 km along the Lower Mekong River, regardless of the type of land use. Water parameters, including temperature, turbidity, DO, and air temperature, also differentially affected the abundance of bacterial families along the Mekong River.

Minimizing the risks associated with manure-borne pathogenic microorganisms requires an understanding of microbial survival under realistic field conditions. The objective of this 3-year study was to assess the fate of Salmonella (SALM) and fecal indicator bacteria (FIB), E. coli (EC) and enterococci (ENT), in glacial till-derived soils, after application of poultry manure (PM) to cornfields under chisel-plowed (CP) or no-till (NT) management. From 2010 to 2012, soil samples were obtained each spring at 0–15- and 15–30-cm depths, to determine whether over-wintering of target bacteria had occurred. Sampling was followed by application of PM at low (PM1) and high (PM2) rates, based on nitrogen application goals. In 2012, soil samples were collected 21, 42, and 158 days after manure application (DAM), to assess the effects of time, application rates, and tillage on frequency of detection and concentrations of target bacteria. Despite dry conditions, all three target organisms were detected 158 DAM in 2012, and detection of these organisms in spring soil samples from manured plots in 2011 and 2012, nearly a full year after PM application, suggests that these organisms can persist in the soil environment long after application. The highest SALM concentration (790 cfu/g dry weight) and detection rate (25%) was found in PM2 plots 42 DAM. SALM were detected more frequently in CP plots (20%) compared to NT plots (5%). In contrast, tillage practices had no apparent effect on EC or ENT survival, as indicated by both soil, and decay rates estimated from tile-water bacteria concentrations. Decay rate constants (μ) ranged from 0.044 to 0.065 day⁻¹ for EC and 0.010 to 0.054 day⁻¹ for ENT.

Copper nanoparticle Cu (d = 55 ± 15 nm) and CuO nanoparticles (d = 90 ± 10 nm) were used in the studies (OOO Platina, Russia). Using the method of pure cultures, we extracted Lactobacillus, Enterococcus, and Enterobacterium from the intestines of broilers. Additionally, strains of Bacillus subtilis 10641 and Bifidobacterium were involved in probiotic strains. The data obtained in the course of the study testify to the insignificant biotoxicity of copper nanoparticles with respect to representatives of the genera Lactobacillus (30 to 15 μg/ml) and Bifidobacterium (30 μg/ml), with the most sensitive bacteria being the genus Lactobacillus, for which a concentration of 7.5 μg/ml was subinhibitory. The second stage was the study using method of agar wells. In the course of the experiment, we obtained results confirming the data of the research by the serial dilution method. In this case, as in the first case, the data indicate the insignificant biotoxicity of copper nanoparticles in relation to representatives of the genera Lactobacillus and Bifidobacterium. We have studied the bioaccumulating ability of microorganisms of the studied metals. In all the studies carried out, as in the first series of experiments, representatives of the genera Lactobacillus and Bifidobacterium with the lowest bioaccumulative ability were the most sensitive to copper nanoparticles and were 3.1 and 8.2%, respectively. The use of nanoparticles as a component of the fodder additive in small concentrations does not adversely affect not only the probiotic strains, but also the main representatives of the normoflora (Lactobacillus, Enterococcus, and Enterobacterium) of the poultry, the positive effect of the copper nanoparticles being directly related to low level of dissociation of nanoparticles, since biologically active ions will be released much more slowly, thereby creating a prolonged effect of exposure.