Impact of heavy precipitation events on pathogen occurrence in estuarine areas of the Puzi River in Taiwan.

Pathogen populations in estuarine areas are dynamic, as they are subject to multiple natural and anthropogenic challenges. Heavy rainfall events bring instability to the aquatic environment in estuaries, causing changes in pathogen populations and increased environmental sanitation and public health concerns. In this study, we investigated the effects of heavy precipitation on the occurrence of pathogens in the Puzi River estuary, which is adjacent to the largest inshore oyster farming area in Taiwan. Our results indicated that Vibrio parahaemolyticus and adenovirus were the most frequently detected pathogens in the area. There was a significant difference (Mann-Whitney U test, p < 0.01) in water quality parameters, including total coliform, Escherichia coli, water temperature, turbidity, salinity, and dissolved oxygen, between groups with and without V. parahaemolyticus. In addition, the detection rate was negatively correlated with the average daily rainfall (r2 > 0.8). There was no significant difference between water quality parameters and the presence/absence of adenovirus, but a positive correlation was observed between the average daily rainfall and the detection rate of adenovirus (r2 ≥ 0.75). We conclude that heavy precipitation changes estuarine water quality, causing variations in microbial composition, including pathogens. As extreme weather events become more frequent due to climate change, the potential impacts of severe weather events on estuarine environments require further investigation.

Effect of temperature on opportunistic pathogen gene markers and microbial communities in long-term stored roof-harvested rainwater.

Roof-harvested rainwater (RHRW) has received increasing attention in recent years as an alternative water source for domestic use, yet its biological stability during storage is not fully understood. This study investigated the effects of temperature (4 °C, 20 °C and 30 °C) on the microbiological characteristics of RHRW over a storage period of 60 days by targeting different microbial groups including total bacteria and fecal indictor Escherichia coli, bacterial opportunistic pathogen genera and species (Legionella spp, Legionella pneumophila, Mycobacterium spp, Mycobacterium avium, Pseudomonas aeruginosa), and two amoebas (Acanthamoeba and Vermamoeba vermiformis). The rainwater chemistry demonstrated no obvious change during storage. The highest biomass was observed in RHRW stored at 30 °C, as measured by heterotrophic bacterial counts, adenosine triphosphate, and 16S rRNA gene numbers. Gene markers of E. coli, Legionella spp., P. aeruginosa, and V. vermiformis were detected in fresh RHRW and can persist during RHRW storage; whereas P. aeruginosa was the only species demonstrated significant regrowth at higher storage temperatures (P < 0.05). Acanthamoeba spp. was only detected in RHRW after 50 days of storage at three investigated temperatures, highlighting increased health risks in long-term stored RHRW. Bacterial community compositions were significantly different in RHRW stored at different temperatures, with increased variations among triplicate storage bottles noted at higher temperatures along with storage time. The results provide insights into RHRW storage practices in terms of mitigating microbial contamination risks.

Diversidad de especies de micobacterias no tuberculosas aisladas en ambientes acuáticos de la ciudad de General Pico, La Pampa, Argentina / Species diversity of non-tuberculous mycobacteria isolated from aquatic environments of General Pico city, Province of La Pampa (Argentina)

Las micobacterias no tuberculosas (MNT) no solo se estudian por su importancia como patógenos oportunistas, sino también por sus aplicaciones en biotecnología y biorremediación. Nuestro objetivo fue determinar la presencia de micobacterias en los distintos hábitats acuáticos de la ciudad de General Pico (provincia de La Pampa), así como su diversidad. Los porcentajes de muestras positivas a micobacterias fueron los siguientes: 37,5% en el sistema de distribución de agua de red, 32,6% en el acuífero que abastece dicho sistema, 36,8% en el agua proveniente de las precipitaciones, 53,1% en los humedales del área de influencia, 80% en los natatorios cubiertos y 33,3% en las fuentes decorativas ubicadas en plazas públicas. De los 90 aislamientos de MNT obtenidos el 8,9% no logró ser identificado a nivel de especie con los métodos utilizados, que incluyeron pruebas fenotípicas y métodos moleculares. Las especies más frecuentemente aisladas fueron Mycobacterium fortuitum y Mycobacterium gordonae. Algunas especies identificadas han sido reportadas en casos de micobacteriosis en nuestro país, entre ellas M. fortuitum, M. gordonae, M. intracellulare, M. vaccae, M. lentiflavum y M. nonchromogenicum. No se aislaron MNT en muestras de agua de red con concentraciones de cloro activo residual mayores de 0,8mg/l, mientras que en los natatorios la presencia de hasta 1,5mg/l de cloro activo residual no fue una limitante para la proliferación de estos microorganismos. Se puede considerar que la incidencia de micobacterias en los ambientes acuáticos de General Pico es cercana al 35%, y que la presencia de estos microorganismos y su diversidad se ve afectada por el contacto con el hombre y sus actividades, como así también por la existencia de vida animal.

Non-tuberculous mycobacteria (NTM) are studied not only for their importance as emerging opportunistic pathogens but also for their applications in biotechnology and bioremediation. Our aim was to determine the occurrence and diversity of mycobacteria in different aquatic habitats of General Pico city, Province of La Pampa. The percentage of samples with positive cultures for mycobacteria were the following: 37.5% recovered from the water supply distribution system; 32.6% from the aquifer that supplies water to the distribution system; 36.8% from rain water; 53.1% from the two wetlands in the area of influence; 80% from indoor swimming pools; and 33.3% from water fountains in downtown public squares. Of the 90 NTM isolates, 8.9% could not be identified at the species level with any of the used methods, phenotypic tests and molecular methods. Mycobacterium fortuitum and Mycobacterium gordonae were the most frequently isolated species. Some of the identified species such as, M. fortuitum, M. gordonae, M. intracellulare, M. vaccae, M. lentiflavum and M. nonchromogenicum, have been reported in cases of mycobacteriosis in Argentina. Mycobacteria with values higher than 0.8mg/ml of residual active chlorine were not recovered from the drinking water supply network, whereas in the swimming pools the presence of up to 1.5 mg/l was not a constraint. Based on our results, the presence of mycobacteria in aquatic environments is close to 35% and their occurrence and diversity is affected both by contact with man and his activities as well as by the existence of animal life.

Evaluation of harvested rainwater quality at primary schools of southwest coastal Bangladesh.

Rainwater is a typical source of drinking water in the coastal areas of Bangladesh given the acute scarcity of drinking water. This study assessed potability of harvested rainwater of primary schools in southwest coastal Bangladesh. Water samples collected from 23 primary schools of Mongla sub-district under Bagerhat district were evaluated for indicator bacteria (total coliform (TC) and E. coli) and physico-chemical parameters (pH, electrical conductivity, turbidity, total dissolved solid, Fe, Zn, Pb, and Cd). Median concentrations of TC and E. coli in the harvested rainwater samples were respectively 3000 cfu/100 ml and 6 cfu/100 ml. However, concentrations of these indicator bacteria were lower at the consumption points which received a prior treatment. Concentration of Pb exceeded the maximum allowable limit for drinking water indicated by WHO and Bangladesh drinking water guideline value in 92% and 61% of the samples respectively, and the mean concentration was 0.08 mg/l (8 times higher than the WHO guideline value). The Pb contamination possibly occurred from the painting on roof railing and roof stair room. Therefore, consumption of harvested rainwater at primary schools may cause substantial health risk for the school-going children.

Relative proportions of E. coli and Enterococcus spp. may be a good indicator of potential health risks associated with the use of roof harvested rainwater stored in tanks.

A total of 285 water samples were collected from 71 roof harvested rainwater tanks from four villages in different provinces over a two-year (2013-2014) period during the early (October to December) and late (January to March) rainy season. Water quality was evaluated based on Escherichia coli, faecal coliforms and Enterococcus spp. prevalence using the IDEXX Quanti-Tray quantification system. Real-Time PCR was used to analyse a subset of 168 samples for the presence of Shigella spp., Salmonella spp. and E. coli virulence genes (stx1, stx2 and eaeA). Escherichia coli were detected in 44.1% of the samples, Enterococcus spp. in 57.9% and faecal coliforms in 95.7%. The most prevalent E. coli concentrations in harvested rainwater were observed in 29.1% of samples and 22.5% for Enterococcus spp. and, were within 1-10 cfu/100 ml and 10-100 cfu/100 ml, respectively, whereas those for faecal coliforms (36.6%) were within 100-1000 cfu/100 ml. On average 16.8% of the samples had neither E. coli nor Enterococcus spp. detected, while 33.9% had only Enterococcus spp. and 23.7% had only E. coli. E. coli and Enterococcus spp. were detected together in 25.5% of the samples. Evaluation of samples for potential pathogenic bacteria showed all tested samples to be negative for the Shigella spp. ipaH gene, while five tested positive for Salmonella ipaB gene. None of the samples tested positive for the stx1 and stx2 genes, and only two tested positive for the eaeA gene. These findings are potentially useful in the development of a simplified risk assessment strategy based on the concentrations of indicator bacteria.

Muddy puddles - the microbiology of puddles located outside tertiary university teaching hospitals.

In the British Isles, the frequency of rain results in the formation of puddles on footpaths and roads in/around hospitals. No data are available demonstrating the microbiological composition of such puddles and therefore a study was undertaken to examine the microbiology of puddles in the grounds of two tertiary university-teaching hospitals (18 sites) and compared with control puddles from non-hospital rural environments (eight sites), estimating (i) total viable count; (ii) identification of organisms in puddles; (iii) enumeration of Escherichia coli: (iv) detection of Extended Spectrum ß-Lactamase producing organisms and (v) direct antimicrobial susceptibility testing. A mean count of 2·3 × 103  CFU per ml and 1·0 × 109  CFU per ml was obtained for hospital and non-hospital puddles respectively. Isolates (n = 77; 54 hospital and 23 non-hospital) were isolated comprising of 23 species among 17 genera (hospital sites), where the majority (10/16; 62·5%) of genera identified were Gram-negative approximately, a fifth (20·6%) were shared by hospital and non-hospital rural samples. Escherichia coli was detected in half of the hospital puddles and under-half (37·5%) of the rural puddles extended spectrum ß-lactamase organisms were not detected in any samples examined. Rainwater puddles from the hospital and non-hospital environments contain a diverse range of bacteria, which are capable of causing infections. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the presence of a wide diversity of bacterial taxa associated with rainwater puddles around hospitals, many of which are capable of causing human disease. Of clinical significance is the presence of Pseudomonas aeruginosa isolated from a hospital puddle, particularly for patients with cystic fibrosis. The presence of potentially disease-causing bacteria in puddles in and around hospitals identifies a new potential environmental reservoir of bacteria. Furthermore work is now needed to define their potential of entering or exiting hospital wards by contaminated footwear.

Impact of storm runoff on Salmonella and Escherichia coli prevalence in irrigation ponds of fresh produce farms in southern Georgia.

AIMS: To examine Salmonella and Escherichia coli in storm runoff and irrigation ponds used by fresh produce growers, and compare Salmonella serovars with those found in cases of human salmonellosis. METHODS AND RESULTS: We collected water before and after rain events at two irrigation ponds on farms in southern Georgia, USA, and collected storm runoff/storm flow within the contributing watershed of each pond. Salmonella and E. coli concentrations were higher in ponds after rain events by an average of 0·46 (P < 0·01) and 0·61 (P < 0·05) log10 most probable number (MPN) per 100 ml respectively. Salmonella concentrations in storm runoff from fields and forests were not significantly higher than in ponds before rain events, but concentrations in storm flow from streams and ditches were higher by an average of 1·22 log10 MPN per 100 ml (P < 0·001). Eighteen Salmonella serovars were identified from 155 serotyped isolates, and eight serovars were shared between storm runoff/storm flow and ponds. Seven of the serovars, including five of the shared serovars, were present in cases of human illness in the study region in the same year. However, several serovars most commonly associated with human illness in the study region (e.g. Javiana, Enteritidis, and Montevideo) were not found in any water samples. CONCLUSIONS: Salmonella and E. coli concentrations in irrigation ponds were higher, on average, after rain events, but concentrations of Salmonella were low, and the ponds met FDA water quality standards based on E. coli. Some similarities and notable differences were found between Salmonella serovars in water samples and in cases of human illness. SIGNIFICANCE AND IMPACT OF THE STUDY: This study directly examined storm runoff/storm flow into irrigation ponds and quantified increases in Salmonella and E. coli following rain events, with potential implications for irrigation pond management as well as human health.

Abundance of Naegleria fowleri in roof-harvested rainwater tank samples from two continents.

Roof-harvested rainwater (RHRW) has been used as an alternative source of water in water scarce regions of many countries. The microbiological and chemical quality of RHRW has been questioned due to the presence of bacterial and protozoan pathogens. However, information on the occurrence of pathogenic amoeba in RHRW tank samples is needed due to their health risk potential and known associations with opportunistic pathogens. Therefore, this study aims to determine the quantitative occurrence of Naegleria fowleri in RHRW tank samples from Southeast Queensland (SEQ), Australia (AU), and the Kleinmond Housing Scheme located in Kleinmond, South Africa (SA). In all, 134 and 80 RHRW tank samples were collected from SEQ, and the Kleinmond Housing Scheme, Western Cape, SA, respectively. Quantitative PCR (qPCR) assays were used to measure the concentrations of N. fowleri, and culture-based methods were used to measure fecal indicator bacteria (FIB) Escherichia coli (E. coli) and Enterococcus spp. Of the 134 tank water samples tested from AU, 69 and 62.7% were positive for E. coli, and Enterococcus spp., respectively. For the SA tank water samples, FIB analysis was conducted for samples SA-T41 to SA-T80 (n = 40). Of the 40 samples analyzed from SA, 95 and 35% were positive for E. coli and Enterococcus spp., respectively. Of the 134 water samples tested in AU, 15 (11.2%) water samples were positive for N. fowleri, and the concentrations ranged from 1.7 × 102 to 3.6 × 104 gene copies per 100 mL of water. Of the 80 SA tank water samples screened for N. fowleri, 15 (18.8%) tank water samples were positive for N. fowleri and the concentrations ranged from 2.1 × 101 to 7.8 × 104 gene copies per 100 mL of tank water. The prevalence of N. fowleri in RHRW tank samples from AU and SA thus warrants further development of dose-response models for N. fowleri and a quantitative microbial risk assessment (QMRA) to inform and prioritize strategies for reducing associated public health risks.

Presence of microbial and chemical source tracking markers in roof-harvested rainwater and catchment systems for the detection of fecal contamination.

Microbial source tracking (MST) and chemical source tracking (CST) markers were utilized to identify fecal contamination in harvested rainwater and gutter debris samples. Throughout the sampling period, Bacteroides HF183 was detected in 57.5 % of the tank water samples and 95 % of the gutter debris samples, while adenovirus was detected in 42.5 and 52.5 % of the tank water and gutter debris samples, respectively. Human adenovirus was then detected at levels ranging from below the detection limit to 316 and 1253 genome copies/µL in the tank water and debris samples, respectively. Results for the CST markers showed that salicylic acid (average 4.62 µg/L) was the most prevalent marker (100 %) in the gutter debris samples, caffeine (average 18.0 µg/L) was the most prevalent in the tank water samples (100 %) and acetaminophen was detected sporadically throughout the study period. Bacteroides HF183 and salicylic acid (95 %) and Bacteroides HF183 and caffeine (80 %) yielded high concurrence frequencies in the gutter debris samples. In addition, the highest concurrence frequency in the tank water samples was observed for Bacteroides HF183 and caffeine (60 %). The current study thus indicates that Bacteroides HF183, salicylic acid and caffeine may potentially be applied as source tracking markers in rainwater catchment systems in order to supplement fecal indicator analyses.

The life history of the plant pathogen Pseudomonas syringae is linked to the water cycle.

Pseudomonas syringae is a plant pathogen well known for its capacity to grow epiphytically on diverse plants and for its ice-nucleation activity. The ensemble of its known biology and ecology led us to postulate that this bacterium is also present in non-agricultural habitats, particularly those associated with water. Here, we report the abundance of P. syringae in rain, snow, alpine streams and lakes and in wild plants, in addition to the previously reported abundance in epilithic biofilms. Each of these substrates harbored strains that corresponded to P. syringae in terms of biochemical traits, pathogenicity and pathogenicity-related factors and that were ice-nucleation active. Phylogenetic comparisons of sequences of four housekeeping genes of the non-agricultural strains with strains of P. syringae from disease epidemics confirmed their identity as P. syringae. Moreover, strains belonging to the same clonal lineage were isolated from snow, irrigation water and a diseased crop plant. Our data suggest that the different substrates harboring P. syringae modify the structure of the associated populations. Here, we propose a comprehensive life cycle for P. syringae--in agricultural and non-agricultural habitats--driven by the environmental cycle of water. This cycle opens the opportunity to evaluate the importance of non-agricultural habitats in the evolution of a plant pathogen and the emergence of virulence. The ice-nucleation activity of all strains from snow, unlike from other substrates, strongly suggests that P. syringae plays an active role in the water cycle as an ice nucleus in clouds.

Confirmation of putative stormwater impact on water quality at a Florida beach by microbial source tracking methods and structure of indicator organism populations.

The effect of a stormwater conveyance system on indicator bacteria levels at a Florida beach was assessed using microbial source tracking methods, and by investigating indicator bacteria population structure in water and sediments. During a rain event, regulatory standards for both fecal coliforms and Enterococcus spp. were exceeded, contrasting with significantly lower levels under dry conditions. Indicator bacteria levels were high in sediments under all conditions. The involvement of human sewage in the contamination was investigated using polymerase chain reaction (PCR) assays for the esp gene of Enterococcus faecium and for the conserved T antigen of human polyomaviruses, all of which were negative. BOX-PCR subtyping of Escherichia coli and Enterococcus showed higher population diversity during the rain event; and higher population similarity during dry conditions, suggesting that without fresh inputs, only a subset of the population survives the selective pressure of the secondary habitat. These data indicate that high indicator bacteria levels were attributable to a stormwater system that acted as a reservoir and conduit, flushing high levels of indicator bacteria to the beach during a rain event. Such environmental reservoirs of indicator bacteria further complicate the already questionable relationship between indicator organisms and human pathogens, and call for a better understanding of the ecology, fate and persistence of indicator bacteria.

Resistência a antibióticos e a metais pesados de Escherichia coli isoladas de agua do mar e galerias pluviais / Resistance to antibiotics and heavy metals from Escherichia coli isolated from sea water and pluvial galeries

OBJECTIVE: Test the resistance to antibiotics and heavy metals of E. coli strains isolated from storm sewer water and adjacent seawater samples from three beaches (Meio, Area Preta and Ponta Negra) in the city of Natal/RN/Brazil, and determine the association among those characteristics. METHODS: A total of 98 strains of E. coli, 50 from storm sewers and 48 from the seawater were analyzed resistance to several antimicrobials by disk diffusion and agar dilution and to heavy metals by dilution in plates with aqueous solutions of CuSO4 incorporated to Mueller Hinton agar in concentrations of 100, 150, 200 and 250 ig/mL and HgCl2 in concentrations of 5, 10, 20, 30, and 50 ig/mL. Standard strains were used as control. RESULTS: Among the twelve antimicrobials tested, 28 (28.5%) of E. coli strains showed resistance to different antimicrobials drugs to seven. The greatest resistance rate was to tetracycline (46.4%), ampicillin (39.3%) and cephalothin (32.1%), with the remainder (nitrophurantoine, nalidixic acid, sulfatomexazol-trimethoprin and chloramphenicol) at lower percentages. Among the heavy metals, all the strains (100%) were resistant to zinc and to copper in the largest concentration (250 ig/mL), and 18.4% were resistant to HgCl the 50 ig/mL. Ten (55.5%) of the E. coli strains resistant to Hg were associated to resistance to antibiotics. CONCLUSION: These results suggest the existence of extra chromosomal genes in E. coli strains isolated from storm sewer water and adjacent seawater, which encoders of the resistance to antibiotics and heavy metals.