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The Ria Formosa, Southern Portugal (Algarve) is a mesotidal lagoon with restricted exchange with Atlantic Ocean. Due to arid Mediterranean climate and absence of large freshwater inputs, the lagoon remains hypersaline relative to adjacent ocean (≥37 psu). Generally, fecal coliforms and land-derived bacteria do not survive long in seawater due to UV, osmotic shock, oligotrophy and competition with marine bacteria. However, survival of human pathogenic viruses in natural waters remains relatively unknown. In view of 2020–2021 SARS-CoV-2 pandemic, this preliminary study aimed to estimate potential contamination by wastewater and persistence in marine environment. Five sampling campaigns (45 total samples) were completed in 2020 and 2021 at three stations in the lagoon, as well as inflow and outflow from three wastewater treatment plants (WWTPs). Despite relatively high fecal contamination at lagoon stations, SARS-CoV-2 could not be detected in surface waters using improved methodology. Untreated wastewaters were positive at Faro-Aeroporto 21/10/2020, 14/12/2020, 08/03/2021; Vilamoura 21/10/2020, 14/12/2020; Olhão 14/12/2020. All treated wastewaters were negative, as well as environmental samples. Highest viral titers were observed in Faro-Aeroporto WWTP on 8 March 2021 (1.35 x 105 genomic copies L−1) coinciding with peak SARS-CoV-2 cumulative positive cases in Algarve (2.03 x 104), yet SARS-CoV-2 was not detected in Vilamoura and Olhão raw wastewater then. Thus, the contamination potential of SARS-CoV-2 seemed non-existent in the Ria Formosa during peak pandemic surges in Algarve. However, predicted climate change and its impact on microbial populations remains a challenge to be addressed by both health and tourist authorities.

Seaweed from Greenland has potential as a food source. However, human sewage is discharged directly to the sea in the vicinity of the communities, a practice which could lead to the seaweed becoming contaminated by human pathogens. The objective of this study was to investigate the effect of wastewater discharge on the bacterial communities of wild populations of Fucus sp. sampled in the tidal zones in the vicinity of Sarfannguit, a smaller settlement (∼110 inhabitants) with limited discharge, and Sisimiut, Greenland's second biggest town (∼5500 inhabitants). Fecal indicator bacteria (coliforms, Escherichia coli and the human fecal molecular marker HF183 [Fucus sp. only]) were consistently detected on Fucus sp. and in seawater from Sisimiut. In contrast, coliforms and E. coli were only detected once in samples collected close to the waste dump site in Sarfannguit. Presence of fecal indicator bacteria in seawater and on Fucus sp. coincided, indicating the utility of surveying seawater. MALDI-TOF mass spectrometry analysis of bacterial isolates identified fish and human pathogens on seaweed from Sisimiut while >80% of isolates from Sarfannguit could not be identified using existing data bases. Amplicon sequence variants belonging to Rhodobacteraceae and Flavobacteriaceae were dominant families in all Fucus sp. samples. However, wastewater discharge effected major changes in the overall composition of the seaweed microbiota as evidenced by analysis of the beta diversity. In conclusion, the microbiota on Fucus sp. harvested in the intertidal zones in a small, relatively unimpacted community, and close to the wastewater discharge of a larger community showed marked differences and the presence of human pathogens on sewage impacted Fucus sp. from the large community. It is recommended that microbiological criteria and guidelines regarding suitable seaweed harvest and cultivation sites be established, especially considering potential sources of anthropogenic impact on the local marine environment.

In this study, the influence of the spatial arrangement of shallow wells and pit latrines on water quality was evaluated in clayey soil during dry and rainy seasons, using 123 randomly selected wells. The distance between well and the nearest latrine was measured and the location of the well in the yard was characterized. The colony forming units (CFU/100 mL) of fecal coliforms were quantified, and pH, electrical conductivity (CE) and turbidity were measured. 100% of the wells were located less than 25 m from the latrine, 74.8% were located in the middle of the yard. In the dry season, 42.4% of the samples presented up to 12 CFU/100 mL, and in the rainy season, 84.4% presented up to 139 CFU/100 mL. 56.9% had pH values between 6.5 and 8.5, and 63.4% presented EC values between 50 and 571 μS/cm. In the dry season, 40.7% of the samples had values below 5 NTU, and 59.3% up to 50 NTU. In the rainy season, 86.6% had values between 6 and 300 NTU. Pearson's correlation between all variables was weak. The wells are susceptible to high fecal contamination, although the clayey soil seems to mitigate the expected high levels of microbial contamination. HIGHLIGHTS 100% of the wells are less than 25 meters from the nearest latrine.; In 57.6% of the dry season samples, bacterial CFU/100 mL were not detected.; In the rainy season, 84.4% of the samples were positive for bacterial CFU/100 mL.; The clay soil seems to mitigate the expected high levels of bacterial contamination.

BackgroundHousehold-level sanitation interventions have had limited effects on child health or environmental contamination, potentially due to low community coverage. Higher community-level coverage with safely managed sanitation can reduce opportunities for disease transmission.MethodsWe estimated associations between community sanitation coverage, environmental fecal contamination, and child health among 360 compounds in the control arm of the WASH Benefits trial in rural Bangladesh (NCT01590095). In each compound, we enumerated E. coli in environmental samples and recorded the 7-day prevalence of caregiver-reported diarrheal disease and acute respiratory infections (ARI) in children under five. We observed indicators of latrine access and quality among all neighboring compounds within 100m of study compounds. We defined community coverage as the proportion of neighboring compounds with (1) at least one latrine, and (2) exclusively hygienic latrines (improved facility observed to safely contain feces), within both 50m and 100m of study compounds. We assessed effect modification by population density and season.ResultsAdjusted for confounders, study compounds surrounded by 100% coverage of at least one latrine per compound within 50m had slightly lower log10E. coli counts in stored water (Δlog=-0.13, 95% CI -0.26, -0.01), child hand rinses (Δlog=-0.13, 95% CI -0.24, -0.02), and caregiver hand rinses (Δlog=-0.16, 95% CI -0.29, -0.03) and marginally lower prevalence of diarrheal disease (prevalence ratio [PR]=0.82, 95% CI 0.64, 1.04) and ARI (PR=0.84, 95% CI 0.69, 1.03) compared to compounds surrounded by

Quantifying the contribution of individual exposure pathways to a child's total ingestion of fecal matter could help prioritize interventions to reduce environmental enteropathy and diarrhea. This study used data on fecal contamination of drinking water, food, soil, hands, and objects and second-by-second data on children's contacts with these environmental reservoirs in rural Bangladesh to assess the relative contribution of different pathways to children's ingestion of fecal indicator bacteria and if ingestion decreased with the water, sanitation, and hygiene interventions implemented in the WASH Benefits Trial. Our model estimated that rural Bangladeshi children

Use of antibiotics inevitably leads to antimicrobial resistance. Selection for resistance occurs primarily within the gut of humans and animals as well as in the environment through natural resistance and residual antibiotics in streams and soil. We evaluated antimicrobial resistance in Gram negative bacteria from a river system in a rural community in Bahia, Brazil. Water was collected from the Jiquiriçá and Brejões rivers and the piped water supply. Additionally, stools were collected from a random sample of residents, cows, pigs and horses near the river. The samples were screened for bacteria resistant to ciprofloxacin, cefotaxime, and meropenem and identified biochemically at the genus and species levels. Microbial source tracking demonstrated that ruminant and human fecal contamination increased as the rivers neared the village center and decreased after the last residence. Antibiotic bacteria were identified from all samples (n = 32). No bacteria were resistant to carbapenems, but the majority of the enterobacteria were resistant to ciprofloxacin, even though this class of antibiotics is not commonly used in food animals in this region. Considering these facts, together with the pattern of human fecal contamination, a human source was considered most likely for these resistant isolates. Resumo O uso de antibióticos inevitavelmente leva à resistência antimicrobiana. A seleção para resistência antimicrobiana ocorre principalmente no intestino de seres humanos e animais, bem como no meio ambiente, através da resistência natural e resíduos de antibióticos nos esgotos e no solo. Avaliamos a resistência antimicrobiana em bactérias Gram-negativas de um sistema fluvial em uma comunidade rural da Bahia, Brasil. A água foi coletada nos rios Jiquiriçá e Brejões e no abastecimento de água encanada. Além disso, foram coletadas amostras randomizadas de fezes de moradores, vacas, porcos e cavalos próximos ao rio. As amostras foram triadas para bactérias resistentes à ciprofloxacina, cefotaxima e meropenem e identificadas bioquimicamente nos níveis de gênero e espécie. O rastreamento de fontes microbianas demonstrou que a contaminação fecal de ruminantes e humanos aumentou à medida que os rios se aproximavam do centro da vila e diminuía após a última residência. Bactérias resistentes a antibióticos foram identificadas em todas as amostras (n = 32). Nenhuma bactéria demonstrou ser resistente aos carbapenêmicos testados, contudo, foi encontrado enterobactérias resistentes à ciprofloxacina, ainda que essa classe de antibióticos não seja comumente usada na medicina veterinária dos animais dessa região. Considerando esses fatos, juntamente com o padrão de contaminação fecal avaliado, a fonte de contaminação humana foi considerada a mais provável na interação desses isolados resistentes.

Wastewater treatment generates a huge amount of sewage sludge, which is a source of environmental pollution. Among the alternatives for the management of this waste, industrial composting stands out as one of the most relevant. The objective of this study was to analyze the bacterial population linked to this process and to determine its effectiveness for the reduction, and even elimination, of microorganisms and pathogens present in these organic wastes. For this purpose, the bacteriome and the fecal bacteria contamination of samples from different sewage sludge industrial composting facilities were evaluated. In addition, fecal bacteria indicators and pathogens, such as Salmonella, were isolated from samples collected at key stages of the process and characterized for antibiotic resistance to macrolide, β-lactam, quinolone, and aminoglycoside families. 16S rRNA phylogeny data revealed that the process clearly evolved toward a prevalence of Firmicutes and Actinobacteria phyla, removing the fecal load. Moreover, antibiotic-resistant microorganisms present in the raw materials were reduced, since these were isolated only in the bio-oxidative phase. Therefore, industrial composting of sewage sludge results in a bio-safe final product suitable for use in a variety of applications.

We investigated the quality of drinking water and its possible effects on human health in the Dera Ghazi Khan (D. G. Khan) district of Pakistan. Samples were collected from three tehsils of the D. G. Khan district, namely D. G. Khan, Kot Chutta, and Taunsa. A total of 50 samples (n = 50) were collected from the study area using standard procedures. The pH of the water samples ranged from 6.52–8.75, EC 0.31–9.78 dS m−1, and TDS 105–985 mg L−1. The bacterial analysis showed that 9 out of 50 samples (18%) contained pathogenic E. coli bacterial. The results showed that the pH and EC values of some sampling sites exceeded the WHO guidelines for drinking water. It was observed that the pH of only 1 sample, and the EC of 18 samples in D. G. Khan—5 in Kot Chutta and 16 in Tehsil Taunsa—exceeded the WHO guidelines. In terms of E. coli presence and related diseases (hepatitis A, B, and C), we collected data, which were screened and belonged to the sampling sites, from 1378 patients receiving treatment related to hepatitis A, B, and C. It was revealed that 530 patients belonged to the D. G. Khan site, followed by Taunsa (460), and Kot Chutta (388). Based on the results, it was concluded that the quality of drinking water samples generally was good, except for 6% of the samples, assessed using (SAR) and Kelly’s ratio (KR), and 9 sites were positive for E. coli.

Rainfall and associated urban runoff have been linked to an increased deterioration of environmental waters, carrying several pollutants including pathogenic microorganisms. Such happens because fecal matter is washed into storm drainage pipes that are afterward released into environmental waters. Stormwater has not been extensively characterized as it is, because most studies are performed either on drainage pipes that are often impacted by sewage leakage or directly in environmental waters following a rain event. In this study, stormwater collected directly from the streets, was monitored for the presence of fecal indicator bacteria (FIB) and three potential important sources of fecal contamination in urban environments (human, cats, and dogs) in three distinct basins in Lisbon, Portugal. Stormwater was collected in sterilized plastic boxes inserted in the storm drains, therefore collecting only runoff. High concentration of fecal contamination was detected with a high percentage of the samples displayed at least one source of contamination. A strong relationship was found between the number of detected sources and the precipitation levels. Although no statistical correlation was found between the locations and the presence of FIB or source markers, the results show a trend in geographical information on the type of urban use in each basin. To the best of our knowledge, this is the first study analyzing the runoff collected directly from the streets. This study suggests that, in urban areas, stormwater runoff is highly impacted by fecal matter, not only from domestic animals but also from human origin, before any cross-contamination in the drainage system and may, by itself, pose a high risk to human health and the environment, particularly if water reuse of this water without further disinfection treatment is the final goal.

INTRODUCTION: Inadequate wastewater treatment and fecal contamination have a strong environmental impact on antimicrobial resistance (AMR). This study evaluated the profile of AMR enterobacteria and fecal contamination from four surface waters: Jiquiriça-Brejões River and Cabrito, Tororó, and Abaeté Lagoons. METHODS: We analyzed AMR β-lactamase genes using the polymerase chain reaction method and fecal contamination using Coliscan®. RESULTS: We found high levels of fecal contamination, β-lactamase producers, and AMR genes (blaOXA-48, blaSPM, and blaVIM) in all waterbodies. CONCLUSIONS: Poor sanitation evidenced by fecal contamination and human activities around these surface waters contributed to the distribution and increase in AMR enterobacteria.

Drinking water contamination is a frequent problem in developing countries and could be associated with bacterial pathogen carriage in feces. We evaluated the association between the risk of drinking water and bacterial carrier status in children younger than 5 years in a cross-sectional study conducted in 199 households from three Peruvian rural communities. Fecal samples from children were screened for pathogenic Aeromonas, Campylobacter, and Vibrio species, as well as for Enterobacteriaceae, including pathogenic Escherichia coli. The drinking water risk was determined using E. coli as an indicator of contamination. Nineteen (9.5%) children were colonized with pathogens and classified as carriers, all without diarrhea symptoms. Of 199 drinking water samples, 38 (19.1%) were classified as very high risk because of high fecal contamination (> 100 E. coli/100 mL). Shared-use water sources, daily washing of containers, and washing using only water were associated with higher prevalence of bacterial carriage, whereas there was no association between households reporting boiling and chlorination of water and carrier status. The prevalence of carriage in children exposed to very high-risk water was 2.82 (95% CI: 1.21-6.59) times the prevalence of those who consumed less contaminated water, adjusted by the water source and daily washing. Our results suggest that household drinking water plays an important role in the generation of carriers with diarrheal pathogens. Our findings also highlight the importance of interventions to ensure the safety of drinking water. Further studies are needed to validate the observed association and determine its significance with respect to diarrhea in the community.

Infectious disease transmission is frequently mediated by the environment, where people's movements through and interactions with the environment dictate risks of infection and/or illness. Capturing these interactions, and quantifying their importance, offers important insights into effective interventions. In this study, we capture high time-resolution activity data for twenty-five Vietnamese farmers during collection and land application of human excreta for agriculture. Although human excreta use improves productivity, the use increases risks of enteric infections for both farmers and end users. In our study, the activity data are integrated with environmental microbial sampling data into a stochastic-mechanistic simulation of E. coli contamination on hands and E. coli ingested. Results from the study include frequent and variable contact rates for farmers' hands (from 34 to 1344 objects contacted per hour per hand), including highly variable hand-to-mouth contact rates (from 0 to 9 contacts per hour per hand). The frequency of hand-to-mouth contacts was substantially lower than the widely-used frequency previously reported for U.S. Office Workers. Environmental microbial contamination data highlighted ubiquitous E. coli contamination in the environment, including excreta, hands, toilet pit, handheld tools, soils, surfaces, and water. Results from the simulation suggest dynamic changes in E. coli contamination on hands, and wide variation in hand contamination and E. coli ingested amongst the farmers studied. Sensitivity analysis suggests that E. coli contamination on hands and ingested doses are most influenced by contamination of handheld tools, excreta, and the toilet pit as well as by frequency of hand-to-mouth contacts. The study findings are especially relevant given the context: no farmers reported adequate storage time of human excreta, and personal protective mask availability did not prevent hand-to-mouth contacts. Integrating high time-resolution activity data into exposure assessments highlights variation in exposures amongst farmers, and offers greater insight into effective interventions and their potential impacts. (C) 2018 The Authors. Published by Elsevier B.V.

The feather corticosterone concentration (fCORT) is increasingly used to assess long-term stress in birds as this indicator provides the potential to retrospectively evaluate the adrenocortical activity of a bird during the growth period of a feather over several weeks in one sample. However, there is still a lack of knowledge concerning external factors which can potentially influence fCORT in birds. The aim of the present study was to determine whether fCORT in laying hens is affected by previous fecal contamination of the plumage. Fully grown primaries 5 (P5s) of laying hens (n = 40) were used for the study. To test the effects of fecal contamination on fCORT, freshly defecated droppings from laying hens were collected and mixed manually. In order to simulate practical and at the same time standardized contamination, the upper surface of the previously determined middle third of the right P5 of each hen then was evenly coated with the paste prepared from fresh feces. The treated feathers were stored for 24 h protected from light at room temperature under a film cover to prevent evaporation. Thereafter, the applied layer of excreta was removed manually. Contralateral P5s of the same individuals were stored under identical conditions and served as controls. Both treatment and control feathers were washed in an aqueous soap solution, rinsed several times in pure water and air-dried subsequently. After pulverization and methanolic extraction, fCORT was analyzed by ELISA. The mean fCORT of treatment P5s (reference ‘feather length’: 12.88 ± 3.16 pg/mm; reference ‘feather weight’: 7.81 pg/mg ± 1.86 pg/mg) were significantly higher (P < 0.001) than those of control P5s (reference ‘feather length’: 9.76 ± 2.42 pg/mm; reference ‘feather weight’: 5.93 ± 1.44 pg/mg). Our results show that previous contamination with excreta can increase fCORT, which was detectable even after a washing procedure prior to analysis. In conclusion, fecal contamination of feathers is a significant influencing factor that has to be considered when applying fCORT measurements.

Water is essential for the life, but many people lack the accessibility to clean and healthy drinking water and die as a consequence of water-borne infections. Microorganism-mediated water pollution is considered as one of the great concerns to the aquatic environment across the globe. The effluent of fecal matter, hospitals, industry, and cattle farms increase the bacterial load in a water body. Coliform groups of bacteria have long been typically applied as an indicator organism of microbial contamination of the water and historically led to the public health security perception. Among the coliform, Escherichia coli is the indicator of fecal contamination. The multiple tube fermentation technique has been applied as a conventional way to detect coliform in water samples through the fermentation of lactose sugar with production of acid and gas. The potability of water has been measured by the absence or presence of coliform bacteria within the permissible limit referencing the most probable number index value (MPN/100 ml). As fecal pollution indicators, fecal streptococci and Clostridium perfringens are widely used as an alternative to coliform bacteria and have been confirmed via esculin hydrolyzing or catalase-negativity and sulfite reduction tests. Molecular (PCR-based) and enzymatic methods have been applied as a rapid way to detect indicators and other enteric isolates in water samples. Apart from that standard plate count (SPC) of heterotrophic bacteria and biochemical oxygen demand (BOD) techniques also determine the bacterial and organic pollution load in a water sample. Therefore, bacteriological analysis of water indicated that water is polluted by sewage to the extent that it is unsuitable for drinking and also unsuitable for recreation purposes. This is one of the big problems in the twenty-first century is providing everybody with safe drinking or domestic water. The main objective of this article is to highlight the microbial pollution of water with special reference to coliform and its nexus with the environment.

Municipal solid waste and sewage sludge are produced in large quantities that are often managed through industrial composting treatment. Because of their origin, composition, and complexity, ensuring adequate stabilization of the organic matter, and sanitation of fecal contaminants during composting is of the utmost significance, and difficult to achieve on an industrial scale. In this study, the operations of six industrial composting facilities that process municipal solid waste and sewage sludge were evaluated from the point of view of the sanitation achieved and the quality of the compost produced. In addition, the results were compared using the model of industrial compost from green waste. Differences between the plants were ascribable to operations other than composting systems. High phytotoxicity and fecal contamination above legislation thresholds were found in compost produced from municipal solid waste. In contrast, compost from sewage sludge were more stable and mature than those produced from green waste, and also had an adequate level of sanitation. The raw material and operational factors are of great relevance to obtain a stable, mature, and pathogen-free compost.

Alongside efforts to improve safe management of feces along the entire sanitation chain, including after the toilet, global sanitation efforts are focusing on universal access ‘basic’ services: onsite facilities that safely contain excreta away from human contact. Although fecal sludge management is improving in urban areas, open drains remain a common fate for feces in these often densely-populated neighborhoods in low-income countries. To-date, it is unclear to what extent complete coverage of onsite sanitation reduces fecal contamination in the urban environment and how fecal contamination varies within urban drains across neighborhoods by sanitation status within a city. We assessed how neighborhood levels of environmental fecal contamination (via spatially-representative sampling of open drains for E. coli) varied across four neighborhoods with varying income, type and coverage of household sanitation facilities, and population density in Accra, Ghana. Neighborhoods with very high sanitation coverage (≥89%) still had high (>4 log10 CFU/100 mL) E. coli concentrations in drains. Between-neighborhood variation in E. coli levels among the high coverage neighborhoods was significant: drain concentrations in neighborhoods with 93% and 89% coverage (4.7 (95% CI: 4.5, 4.9) & 4.9 (95% CI: 4.5, 5.3) log10 CFU/100 mL, respectively) were higher than in the neighborhood with 97% coverage (4.1 log10 CFU/100 mL, 95% CI: 3.8, 4.4 log10 CFU/100 mL). Compared with the highest coverage neighborhood, the neighborhood with lowest coverage (48%) also had higher E. coli concentrations (5.6 log10 CFU/100 mL, 95% CI: 5.3, 5.9 log10 CFU/100 mL). Although fecal contamination in open drains appeared lower in neighborhoods with higher onsite sanitation coverage (and vice versa), other factors (e.g. fecal sludge management, animals, population density) may affect drain concentrations. These results underscore that neighborhood-level onsite sanitation improvements alone may not sufficiently reduce fecal hazards to public health from open drains. These findings supporting the need for integrated, city-level fecal sludge management alongside multifaceted interventions to reduce fecal contamination levels and human exposure.

This study was performed to improve the ability to predict the concentrations of Escherichia coli in oyster meat and estuarine waters by using environmental parameters, and microbiological and heavy metal contamination from shellfish growing area in southern Thailand. Oyster meat (n = 144) and estuarine waters (n = 96) were tested for microbiological and heavy metal contamination from March 2016 to February 2017. Prevalence and mean concentrations of E. coli were 93.1% and 4.6 × 103 most probable number (MPN)/g in oyster meat, and 78.1% and 2.2 × 102 MPN/100 mL in estuarine water. Average 7-day precipitation, ambient air temperature, and the presence of Salmonella were associated with the concentrations of E. coli in oyster meat (p < 0.05). Raw data (MPN/g of oyster meat and MPN/100 mL of estuarine water) and log-transformed data (logMPN/g of oyster meat and logMPN/100 mL of estuarine water) of E. coli concentrations were examined within two contrasting regression models. However, the more valid predictions were conducted using non-log transformed values. These findings indicate that non-log transformed data can be used for building more accurate statistical models in microbiological food safety, and that significant environmental parameters can be used as a part of a rapid warning system to predict levels of E. coli before harvesting oysters.

Приводятся результаты оценки санитарного состоянии реки Кирпичная, руч. Крутоберега и руч. Кабан, расположенных в пределах г. Петропавловска-Камчатского, по таким санитарнобактериологическими показателями качества водной среды, как общее микробное число и количество бактерий группы кишечной палочки. Установлено, что исследуемые водотоки имеют неудовлетворительное санитарное состояние, обусловленное антропогенным воздействием., The results of the sanitary condition assessment of the Kirpichnaya River, the Krutobereg Stream and the Kaban Stream located within Petropavlovsk-Kamchatsky are given. The sanitary and bacteriological indicators of the aquatic environment quality such as the total microbial number and the number of bacteria of the group of colon bacillus are analized. It is established that the studied watercourses have poor sanitary condition due to the anthropogenic impact.

Currently, around 50% of the world's population lives in towns and cities within 100 km of the coast. Monitoring of viruses that are frequently present in contaminated coastal environments, such as rotavirus (RoV) and norovirus (NoV), which are also the major cause of human viral gastroenteritis, is essential to ensure the safe use of these water bodies. Since exposure to as few as 10–100 particles of RoV or NoV may induce gastrointestinal disease, there is a need to develop a rapid and sensitive diagnostic method for their detection in coastal water samples. In this study, we evaluate the application of methacrylate monolithic chromatographic columns, commercially available as convective interaction media (CIM®), to concentrate pathogenic enteric viruses from saline water samples prior to virus quantification by one-step reverse transcription quantitative PCR (RT-qPCR). Using RoV and NoV as model enteric viruses, we present our results on the most effective viral concentration conditions from saline water matrices using butyl (C4) hydrophobic interaction monolithic support (CIM® C4). C4 monolithic columns exhibit a good capacity to bind both RoV and NoV and both viruses can be eluted in a single step. Our protocol using a 1 ml C4 column enables processing of 400 ml saline water samples in less than 60 min and increases the sensitivity of RoV and NoV detection by approximately 50-fold and 10-fold respectively. The protocol was also scaled up using larger capacity 8 ml C4 columns to process 4000 ml of seawater samples with concentration factors of 300-fold for RoV and 40-fold for NoV, without any significant increase in processing time. Furthermore, C4 monolithic columns were adapted for field use in an on-site application of RoV concentration from seawater samples with performance equivalent to that of the reference laboratory setup. Overall, the results from successful deployment of CIM C4 columns for concentration of rotavirus and norovirus in seawater samples reiterate the utility of monolithic supports as efficient, scalable and modular preparative tools for processing environmental water samples to enhance viral detection using molecular methods.

In the past 20 years, hyperspectral imaging has been widely investigated as an emerging, promising technology for evaluating quality and safety of horticultural products. This technology has originated from remote sensing and joins the domains of machine vision and point spectroscopy to provide superior image segmentation for the detection of defects and contaminations, and to map the chemical composition. Thanks to the advancements in instrumentation and data analysis in the past two decades, hyperspectral imaging technology has evolved into a powerful nondestructive inspection tool and the scope of applications in postharvest quality and safety evaluation has expanded tremendously. In this article, different imaging modes (reflectance, transmittance, fluorescence and Raman) and their combinations, and the potential for real-time acquisition of hyperspectral images at industry relevant speeds are first discussed in terms of their advantages and disadvantages. Next reviewed are different data processing/analysis methods and associated steps from data pre-processing over the spectral and spatial domains to the actual model building and performance evaluation. An overview is then given of hyperspectral imaging applications for external quality and defect evaluation, internal quality and maturity assessment, and food safety detection of horticultural products. Finally, a brief discussion is presented on the challenges and opportunities in future development and application of hyperspectral imaging technology in food quality and safety evaluation of horticultural products.

Understanding household-level transmission pathways of fecal pathogens can provide insight for developing effective strategies to reduce diarrheal illness in low- and middle-income countries. We applied whole bacterial community analysis to investigate pathways of bacterial transmission in 50 rural Bangladeshi households. SourceTracker was used to quantify the shared microbial community in household reservoirs (stored drinking water, soil, and hands) and estimate the percentage of fecal-associated bacteria from child and mothers’ feces in these reservoirs. Among the reservoirs studied, most bacterial transfer occurred between mothers’ and children’s hands and between mothers’ hands and stored water. The relative percentage of human fecal-associated bacteria in all household reservoirs was low. We also quantified the number of identical amplicon sequence variants within and between individual households to assess bacterial community exchange in the domestic environment. Intra-household sharing of bacteria between mothers’ and children’s hands and between hands and soil was significantly greater than inter-household sharing., Graphical abstract Image 1, Highlights • 16S rRNA gene sequencing was used to examine bacterial communities in households. • Low levels of fecal contamination were found on hands, in soil, and stored water. • High proportion of bacteria was shared between mothers and their children.

Wastewater contains elevated concentrations of fecal indicator bacteria (FIB). The type of wastewater treatment technology and septic system density may influence the FIB concentration and exports at the watershed scale. The goal of this study was to gain a better understanding of FIB concentrations and exports from watersheds served by conventional septic (CS) systems, sand filter (SF) septic systems, and a municipal sewer (SEW) system. Seven watersheds (3 CS, 3 SF, and 1 SEW) were monitored to quantify FIB concentration and export monthly from April 2015 to March 2016. The type of wastewater treatment did not yield significant differences in FIB concentration or exports when pooling watersheds using similar wastewater treatment. Watersheds with the highest septic densities (approximately 0.4 systems ha&minus, 1) contained greater FIB concentrations and exports than watersheds with the lowest (approximately 0.1&ndash, 0.2 systems ha&minus, 1), but only FIB concentrations significantly differed. These findings suggest that when the septic system density exceeds 0.4 systems ha&minus, 1, water quality degradation from septic leachate may be observable at the watershed scale, especially in watersheds dominated by residential development. More research is recommended to determine if this density threshold is similar for other water pollutants and/or in watersheds with differing hydrogeological, land use, and wastewater characteristics.

In this research work, a comparative analysis of the legal framework for water reuse in Brazil and Portugal within their scenarios of sanitation and water resources management is presented. It appears that the coverage in drinking water and sanitation services is quite different between the two countries. However, the respective rates of reuse of effluents in relation to the total wastewater treated are similar (1.5% in Brazil and 1.2% in Portugal). Water reuse has become an important alternative source of increasing water availability for sustainable development. Despite the legal aspects of both countries presenting similar management instruments, Brazil tends to be more flexible in relation to the standards of fecal contamination indicators for the reuse of water for irrigation of urban environments, firefighting, and street washing. In the case of reuse for irrigation of agricultural environments and flushing the toilet, legal standards are equally restrictive. Standards for use in civil construction and clearing wastewater galleries are only included in the Brazilian regulatory document., No presente artigo, apresenta-se uma análise comparativa do enquadramento regulatório de reúso de água do Brasil e de Portugal, inserido nos seus cenários de gestão de saneamento ambiental e recursos hídricos. Verifica-se que os índices de atendimento em serviços de água e esgotos são bastante distintos nos dois países, sendo semelhantes, no entanto, as respetivas taxas de reúso de água em relação ao esgoto total tratado (1,5% no Brasil e 1,2% em Portugal). A água de reuso vem se tornando uma importante fonte alternativa de aumento da disponibilidade hídrica para o desenvolvimento sustentável. Apesar dos aspectos de regulamentação de ambos os países apresentarem instrumentos de gestão similares, o Brasil tende a ser mais flexível em relação aos padrões de indicadores de contaminação fecal para o reúso de água para irrigação/rega de ambientes urbanos, combate a incêndio e lavagem de ruas. No caso de reúso para irrigação/rega de ambientes agrícolas e descarga de vaso sanitário, os padrões legais são igualmente restritivos. Padrões para usos na construção civil e na desobstrução de galerias de águas residuais somente constam no documento regulador brasileiro

Field trials were conducted in July–August and October 2012 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, California. One-half milliliter of rabbit, chicken, or pig fecal slurry, containing an average of 4.05 × 107 CFU E. coli O157:H7 (C0), was inoculated onto the upper (adaxial) surface of a lower leaf on 288 heads of lettuce per trial immediately following a 2.5 h irrigation event. To estimate the bacterial inactivation rate as a function of time, fecal matrix, irrigation and seasonal climate effects, sets of lettuce heads (n = 28) were sampled each day over 10 days and the concentration of E. coli O157:H7 (Ct) determined. E. coli O157:H7 was detected on 100% of heads during the 10-day duration, with concentrations ranging from ≤340 MPN/head (∼5-log reduction) to >3.45 × 1012 MPN/head (∼5-log growth). Relative to C0, on day 10 (Ct = 12) we observed an overall 2.6-log and 3.2-log mean reduction of E. coli O157:H7 in July and October, respectively. However, we observed relative maximum concentrations due to bacterial growth on day 6 (maximum Ct = 8) apparently stimulated by foliar irrigation on day 5. From this maximum there was a mean 5.3-log and 5.1-log reduction by day 10 (Ct = 12) for the July and October trials, respectively. This study provides insight into the inactivation and growth kinetics of E. coli O157:H7 on romaine lettuce leaves under natural field conditions. This study provides evidence that harvesting within 24 h post irrigation has the potential to increase the concentration of E. coli O157:H7 contamination, if present on heads of romaine lettuce; foliar irrigation can temporarily stimulate substantial regrowth of E. coli O157:H7.

Microbial source tracking (MST) analysis is essential to identifying and mitigating the fecal pollution of water resources. The signature-based MST method uses a library of sequences to identify contaminants based on operational taxonomic units (OTUs) that are unique to a certain source. However, no clear guidelines for how to incorporate OTU overlap or natural variation in the raw water bacterial community into MST analyses exist. We investigated how the inclusion of bacterial overlap between sources in the library affects source prediction accuracy. To achieve this, large-scale sampling – including feces from seven species, raw sewage, and raw water samples from water treatment plants – was followed by 16S rRNA amplicon sequencing. The MST library was defined using three settings: (i) no raw water communities represented; (ii) raw water communities selected through clustering analysis; and (iii) local water communities collected across consecutive years. The results suggest that incorporating either the local background or representative bacterial composition improves MST analyses, as the results were positively correlated to measured levels of fecal indicator bacteria and the accuracy at which OTUs were assigned to the correct contamination source increased fourfold. Using the proportion of OTUs with high source origin probability, underpinning a contaminating signal, is a solid foundation in a framework for further deciphering and comparing contaminating signals derived in signature-based MST approaches. In conclusion, incorporating background bacterial composition of water in MST can improve mitigation efforts for minimizing the spread of pathogenic and antibiotic resistant bacteria into essential freshwater resources.

Various waterborne pathogens originate from human or animal feces and may cause severe gastroenteric outbreaks. Bacteroides spp. that exhibit strong host- or group-specificities are promising markers for identifying fecal sources and their origins. In the present study, 240 water samples were collected from two major aquaculture areas in Republic of Korea over a period of approximately 1 year, and the concentrations and occurrences of four host-specific Bacteroides markers (human, poultry, pig, and ruminant) were evaluated in the study areas. Host-specific Bacteroides markers were detected widely in the study areas, among which the poultry-specific Bacteroides marker was detected at the highest concentration (1.0–1.2 log10 copies L−1). During the sampling period, high concentrations of host-specific Bacteroides markers were detected between September and December 2015. The host-specific Bacteroides marker-combined geospatial map revealed the up-to-downstream gradient of fecal contamination, as well as the effects of land-use patterns on host-specific Bacteroides marker concentrations. In contrast to traditional bacterial indicators, the human-specific Bacteroides marker correlated with human specific pathogens, such as noroviruses (r=0.337; P

Made available in DSpace on 2019-10-06T15:18:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-01-10 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The objective of this study was to verify the presence of antimicrobial resistant strains of Escherichia coli in pig farms and to use it as a biomarker to evaluate phenotypic and genotypic profiles of antimicrobial susceptibility, as well as the presence of Extended Spectrum Beta-lactamases (ESBLs) and fluoroquinolone resistance genes. Several samples (n = 306) collected from swine farms (n = 100) of Southern Brazil were used for E. coli isolation: 103 of swine feces, 105 of water, and 98 of soil. E. coli isolates were submitted to the disk-diffusion test to verify their antimicrobial susceptibility, to disk-approximation test to detect ESBL-producers, and to PCR analysis to search for ESBLs genes (blaCTY-M2, blaSHV-1, blaTEM-1, blaCTX-M2, blaOXA-1, blaPSE-1) and quinolone resistance genes (qnrA, qnrB and qnrS). The percentage of E. coli isolates found in feces, water and soil samples was 66.02%, 30.48% and 35.71%, respectively. The highest percentages of resistance were obtained for sulfamethoxazole associated with trimethoprim (63.70%), colistin (45.19%) and enrofloxacin (39.26%). Regarding the levels of multidrug resistance, 37.04% of the isolates were resistant to three or more classes of antimicrobials. The most common profile (16%) of multirresistance was GEM-SUT-ENO-COL. The index of multiple resistance to antimicrobials (IRMA) was above 0.2 in 78% of the multiresistant isolates. Out of 135 E. coli isolates, 7.41% was ESBL-producers, of which 50% showed the blaCMY-M2 gene, 40% the blaTEM-1 and 70% the qnrS gene. Of non-ESBL-producing strains resistant to enrofloxacin, 13.04% were positives for qnrS gene. These results demonstrated the presence of fecal contamination in the environment, in addition to high resistance indexes for several antimicrobials, including beta-lactams and fluoroquinolones, which was confirmed by the genetic detection of ESBLs and qnr genes. Post Graduation Program in Animal Science State University of Santa Catarina (UDESC) Western Center of Education Graduate in Animal Science State University of Santa Catarina (UDESC) Western Center of Education Department of Animal Science State University of Santa Catarina (UDESC) Western Center of Education Department of Mathematics Paulista State University Júlio de Mesquita Filho (UNESP) Department of Mathematics Paulista State University Júlio de Mesquita Filho (UNESP)

In recent decades, quantitative microbial risk assessment (QMRA) has been widely used to assess exposure to fecal microbes and associated health risks. In this study, a multipathway exposure assessment model was developed to evaluate exposure to fecal microbes for children under 5 in highly contaminated urban environments. Children had contact with various environmental compartments. The contamination levels of these compartments were estimated from fecal indicator counts in the environmental samples. Structured observations of child behavior (including activities, locations, and time) were used to model behavioral sequences as a dynamic network. The exposure model combines behavior sequences with environmental contamination, using additional exposure factors when needed, to estimate the number of fecal microbes transferred from environmental sources to human oral ingestion. As fecal exposure in a highly contaminated urban environment consists of contributions from multiple pathways, it is imperative to study their relative importance. The model helps us better understand the characteristics of the exposure pathways that may be driven by variation in contamination and by variable behavior, like hygiene and high‐risk activities. Importantly, the model also allows prediction of the quantitative effects of an intervention—the expected reduction in exposure due to infrastructural or behavioral changes—by means of scenario studies. Based on experience with this exposure model, we make specific recommendations for additional studies of child behavior and exposure factors in order to fill critical information gaps and improve the model structure and assumptions.

In this paper, multivariate analysis of sterol concentrations detected in river sediment samples was performed. In order to remove co-dependence of values, concentrations of sterols were transformed using centered log-ratio (CLR) transformation. The main objective of the work was to point out the damaging effects of working in the wrong geometry on the principal component analysis (PCA) assessment of sterol pollution. In order to determine if the dimension lost have effect on the principal component analysis of sterols in sediments, we have performed the PCA using raw and log-ratio transformed sterol data. Additionally, two rounded zero replacement approaches, i.e. a simple-substitution method (DL/2, 0.55DL and DL/√2) and multiplicative replacement strategy (0.65 DL), were compared in order to determine if the replacement values have an effect on PCA results and conclusions. Relevant differences were noted by comparing the results of the principal component analysis obtained with raw data and log-ratio transformed sterol data. Only the PC loadings obtained from the CLR PCA allowed the clear distinction between human-sourced pollution and biogenic sources of sterols, whereas in the case of PCA with raw data loadings were all grouped almost in a single quadrant. For the small proportion of rounded zeros (not more than 10%), two different replacement approaches did not have any effect on transformed PCA output. The results presented in this work have shown that the effect of “closure” in the sterol data can be easily observed from the PCA biplot, and that it obstructs the evaluation of human contribution to pollution of river sediments. Therefore, prior to the PCA, sterol concentrations must be CLR transformed in order to perform a reliable assessment of the sewage contamination.

Empiric quantification of environmental fecal contamination is an important step toward understanding the impact that water, sanitation, and hygiene interventions have on reducing enteric infections. There is a need to standardize the methods used for surface sampling in field studies that examine fecal contamination in low-income settings. The dry cloth method presented in this manuscript improves upon the more commonly used swabbing technique that has been shown in the literature to have a low sampling efficiency. The recovery efficiency of a dry electrostatic cloth sampling method was evaluated using Escherichia coli and then applied to household surfaces in Iquitos, Peru, where there is high fecal contamination and enteric infection. Side-by-side measurements were taken from various floor locations within a household at the same time over a three-month period to compare for consistency of quantification of E. coli bacteria. The dry cloth sampling method in the laboratory setting showed 105% (95% Confidence Interval: 98%, 113%) E. coli recovery efficiency off of the cloths. The field application demonstrated strong agreement of side-by-side results (Pearson correlation coefficient for dirt surfaces was 0.83 (p < 0.0001) and 0.91 (p < 0.0001) for cement surfaces) and moderate agreement for results between entrance and kitchen samples (Pearson (0.53, p < 0.0001) and weighted Kappa statistic (0.54, p < 0.0001)). Our findings suggest that this method can be utilized in households with high bacterial loads using either continuous (quantitative) or categorical (semi-quantitative) data. The standardization of this low-cost, dry electrostatic cloth sampling method can be used to measure differences between households in intervention and non-intervention arms of randomized trials.

International audience; In this study, the capacity of oysters to bioaccumulate fecal stanols and to record a source-specificfingerprint was investigated by the short-term contamination of seawater microcosms containing oysterswith a human effluent. Contaminated oysters bioaccumulated the typical fecal stanols coprostanol and24-ethylcoprostanol and their bioaccumulation kinetics were similar to that of the Fecal Indicator BacteriaEscherichia coli used in European legislation. Although stanol fingerprints of contaminated waterallowed the identification of the human specific fingerprint, this was not the case for oysters. This discrepancyis attributed to (i) high concentrations of endogenous cholestanol and sitostanol, responsiblefor ‘‘unbalanced’’ stanol fingerprints, (ii) different accumulation/depuration kinetics of fecal coprostanoland 24-ethylcoprostanol and (iii) the limits of the analytical pathway used. These results show that fecalstanols bioaccumulated by oysters are useful to record fecal contamination but the usefulness of stanolfingerprints to identify specific sources of contamination in shellfish currently seems limited.

International audience; The objective of this work was to study the effects of washing and purification steps on qualitative and quantitative analysis of fecal stanols in the oyster Crassostrea gigas using either single or a combination of lipid purification steps on silica gel or aminopropyl bonded silica gel (NH2) or a washing step. Among the three analytical pathways compared, the two including water extraction or NH2 purification did not lead to higher recoveries and decreased repeatabilities of extractions compared to the single purification on silica gel. This latter led to similar recoveries (ca. 80 %) and repeatabilities (ca. 10 %) for both spiked standards (coprostanol and sitostanol). This analytical pathway has been applied to oysters collected in a harvesting area in Brittany (France) where fecal contaminations are important and allowed to quantify eight stanols in oysters. The relative proportions of fecal stanols of these oysters were combined with principal component analysis in order to investigate the usefulness of their stanol fingerprints to record a fecal contamination and to distinguish its source between human, porcine and bovine contaminations. Oysters non-fecally contaminated by Escherichia coli did not present specific stanol fingerprints while oysters fecally contaminated had a bovine fingerprint, suggesting a contamination of these samples by bovine sources. As a consequence, the method developed here allows the use of stanol fingerprints of oysters as a microbial source tracking tool that can be applied to shellfish harvesting areas subjected to fecal contaminations in order to identify the different sources of contamination and improve watershed management.

This study was performed to improve the ability to predict the concentrations of Escherichia coli in oyster meat and estuarine waters by using environmental parameters, and microbiological and heavy metal contamination from shellfish growing area in southern Thailand. Oyster meat (n = 144) and estuarine waters (n = 96) were tested for microbiological and heavy metal contamination from March 2016 to February 2017. Prevalence and mean concentrations of E. coli were 93.1% and 4.6 &times, 103 most probable number (MPN)/g in oyster meat, and 78.1% and 2.2 &times, 102 MPN/100 mL in estuarine water. Average 7-day precipitation, ambient air temperature, and the presence of Salmonella were associated with the concentrations of E. coli in oyster meat (p &lt, 0.05). Raw data (MPN/g of oyster meat and MPN/100 mL of estuarine water) and log-transformed data (logMPN/g of oyster meat and logMPN/100 mL of estuarine water) of E. coli concentrations were examined within two contrasting regression models. However, the more valid predictions were conducted using non-log transformed values. These findings indicate that non-log transformed data can be used for building more accurate statistical models in microbiological food safety, and that significant environmental parameters can be used as a part of a rapid warning system to predict levels of E. coli before harvesting oysters.

Field trials were conducted in July–August and October 2012 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, California. One-half milliliter of rabbit, chicken, or pig fecal slurry, containing an average of 4.05 × 107 CFU E. coli O157:H7 (C0), was inoculated onto the upper (adaxial) surface of a lower leaf on 288 heads of lettuce per trial immediately following a 2.5 h irrigation event. To estimate the bacterial inactivation rate as a function of time, fecal matrix, irrigation and seasonal climate effects, sets of lettuce heads (n = 28) were sampled each day over 10 days and the concentration of E. coli O157:H7 (Ct) determined. E. coli O157:H7 was detected on 100% of heads during the 10-day duration, with concentrations ranging from ≤340 MPN/head (∼5-log reduction) to >3.45 × 1012 MPN/head (∼5-log growth). Relative to C0, on day 10 (Ct = 12) we observed an overall 2.6-log and 3.2-log mean reduction of E. coli O157:H7 in July and October, respectively. However, we observed relative maximum concentrations due to bacterial growth on day 6 (maximum Ct = 8) apparently stimulated by foliar irrigation on day 5. From this maximum there was a mean 5.3-log and 5.1-log reduction by day 10 (Ct = 12) for the July and October trials, respectively. This study provides insight into the inactivation and growth kinetics of E. coli O157:H7 on romaine lettuce leaves under natural field conditions. This study provides evidence that harvesting within 24 h post irrigation has the potential to increase the concentration of E. coli O157:H7 contamination, if present on heads of romaine lettuce; foliar irrigation can temporarily stimulate substantial regrowth of E. coli O157:H7.

This study represents the first investigation of microbiological groundwater pollution as a function of aquifer type and season for the Apulia region of southern Italy. Two hundred and seven wells were randomly selected from those monitored by the Regional Agency for Environmental Protection for emergency use. Both compulsory (Escherichia coli, Total Coliform, and Enterococci) and optional (Pseudomonas aeruginosa, Salmonella spp., Heterotrophic Plate Count at 37 and 22 °C) microbiological parameters were assessed regularly at these wells. Groundwater from only 18 of the 207 (8.7 %) wells was potable; these all draw from karst-fissured aquifers. The remaining 189 wells draw from karst-fissured (66.1 %) or porous (33.9 %) aquifers. Of these, 82 (43.4 %) tested negative for Salmonella spp. and P. aeruginosa, while 107 (56.6 %) tested positive for P. aeruginosa (75.7 %), Salmonella spp. (10.3 %), or for both Salmonella spp. and P. aeruginosa (14 %). A logistic regression model shows that the probability of potable groundwater depends on both season and aquifer type. Typically, water samples were more likely to be potable in autumn-winter than in spring-summer periods (odds ratio, OR = 2.1; 95 % confidence interval, 95 % CI = 1.6-2.7) and from karst-fissured rather than porous aquifers (OR = 5.8; 95 % CI = 4.4-7.8). Optional parameters only showed a seasonal pattern (OR = 2.6; 95 % CI = 1.7-3.9). Clearly, further investigation of groundwater microbiological aspects should be carried out to identify the risks of fecal contamination and to establish appropriate protection methods, which take into account the hydrogeological and climatic characteristics of this region.

Poor sanitation infrastructure in rural areas can often lead to high levels of fecal contamination in local waterbodies and subsequent exposure to waterborne disease can occur. Although standard water quality measures such as quantification of E. coli can reveal relative concentrations of fecal contamination, they do not pinpoint the sources of such contamination. Source assessment in rural areas affected by untreated household waste might be improved with the human-specific, microbial source tracking marker HF183. This study attempted to quantify HF183 in two particular Appalachia streams with known discharges of untreated household waste. Water samples were taken above and at multiple points below these discharges on 29 occasions between August 2012 and April 2016, and tested for both HF183 and E. coli. HF183 was detected consistently in one of the study streams, though the concentrations were generally much lower than those previously reported in raw sewage; in the other watershed, HF183 was never detected. Further analysis via a multiple linear regression model showed a positive correlation between the level of E. coli and the proximity and number of known waste discharge points upstream from each sampling site. Primary conclusions of this study include: 1) HF183 is not always detected, even in watersheds with known sources of human fecal contamination, 2) it may be a useful water quality assessment tool where such contamination is suspected, particularly in cases where contaminant source allocation is necessary for setting mitigation priorities. Master of Science

Because the contaminated fresh produce and raw materials for food with animal fecal matter can introduce foodborne illness, it is necessary to develop an automatic inspection system to detect the fecal contamination on fresh produce. The hyperspectral fluorescence imaging system using ultraviolet-A excitation (320∼400 nm) was investigated to detect the bovine fecal contamination on the abaxial and adaxial surfaces of romaine lettuce and baby spinach leaves. An image processing algorithm to detect the fecal contamination spots for public health concern was investigated while it correctly indentifies the clean leaf surfaces. The developed algorithm could successfully detect the fecal contamination spots on the adaxial and abaxial surfaces of romaine lettuce and baby spinach.

This study identified the sources of fecal contamination in the groundwater of different land covers. A total of 300 groundwater samples were collected in the Kathmandu Valley, Nepal, in the dry (n = 152) and wet (n = 148) seasons of 2016. Fecal indicator bacteria were initially enumerated, and then fecal contamination sources were identified using human (BacHum), ruminant (BacR), and pig-associated (Pig2Bac) Bacteroidales quantitative polymerase chain reaction assays. Sixty-six percent (197/300) of the tested groundwater samples had Escherichia coli concentrations higher than the World Health Organization threshold for drinking (&lt, 1 most probable number/100 mL). The fecal contamination of the groundwater was of human (22%, 55/250), ruminant (11%, 28/250), and pig (3%, 8/250) origin. Deep tube wells were less likely to be positive for E. coli and fecal markers compared to shallow dug wells. The human fecal marker was more likely to be detected in sources from built-up as compared to agricultural areas (Adjusted odds ratio (AOR) = 3.60, p = 0.002). Likewise, the ruminant fecal marker was more likely to be detected in sources from agricultural as compared to built-up areas (AOR = 2.90, p = 0.018). These findings suggest the preparation of mitigation strategies for controlling fecal pollution based on land cover and well types.

Molecular methods are useful for both to monitor anthropogenic viral, bacterial, and protozoan enteropathogens, and to track pathogen specific markers in a complex environment in order to reveal sources of these pathogens. Molecular genetic markers for fecal viruses, bacteria, and protozoans hold promise for monitoring environmental pollution and water quality. The demand for microbiologically safe waters grows exponentially due to the global demographic rise of the human population. Economically important shellfish, such as oysters, which are harvested commercially and preferentially consumed raw can be of public health importance if contaminated with human waterborne pathogens. However, feral molluscan shellfish which do not have an apparent economic value serve as indicators in monitoring aquatic environments for pollution with human waterborne pathogens and for sanitary assessment of water quality. Current technology allows for multiplexed species-specific identification, genotyping, enumeration, viability assessment, and source-tracking of human enteropathogens which considerably enhances the pathogen source-tracking efforts.

The aim of this 2-year study was to examine the temporal changes in the concentration of microbiological indicators of water contamination and selected physicochemical parameters within the Bialka river and its selected tributaries in the vicinity of the largest ski station in Bialka Tatrzanska. The study involved 24 series of sampling in eight sites throughout the Bialka river and on its selected tributaries. Temperature, pH, and electrical conductivity (EC25 °C) were measured onsite. The microbiological assays included the numbers of mesophilic and psychrophilic bacteria, Staphylococcus spp. and Salmonella spp., as well as coliforms, thermotolerant (fecal) coliforms, Escherichia coli, Enterococcus faecalis, and sulfate-reducing Clostridium. The chemical analyses were conducted to determine the concentration of NH4 +, NO2 −, NO3 −, and PO4 3−. The analyses showed that contrary to common opinion, waters of the Bialka river are strongly polluted in some sections. Seasonal variation in the prevalence of microbial indicators of fecal pollution was found, and the largest numbers of microorganisms were observed in winter ski season, followed by summer holidays. Similar seasonal pattern was observed in the examined chemical parameters. There was also very strong spatial variation within the tested microbial and chemical parameters, indicating the presence of pollution hotspots in the course of the Bialka river. The employed principal component analysis revealed the presence of two main pollution sources, mainly affecting the quality of river water, i.e., fecal contamination of human origin and the natural source in the form of surface runoff and soil leaching. These factors, depending on the location of the sampling site, occurred in different configurations.

European legislation has defined as process hygiene criteria for the main livestock species (cattle, sheep, goats, horses and pigs) the monitoring of aerobic colony count and Enterobacteriaceae. Detected values above the defined criteria require an improvement in slaughter hygiene and the review of process control. The main source of microbiological contamination of beef carcasses along the slaughterline is of fecal origin, therefore Escherichia coli and Enterobacteriaceae seem to be the most suitable indicators to assess the hygienic status of the slaughter process. Although microbiological criteria addressing indicator bacteria have been in place in industrialized countries for several years, scattered information still exists on factors affecting their counts on beef carcasses along the slaughterline. Therefore, a systematic literature review, covering the period 2000–2012, was conducted to gather information concerning: 1) counts of E. coli and Enterobacteriaceae on beef carcasses linked to different stages of the slaughterline; 2) factors influencing presence/counts of E. coli and Enterobacteriaceae on beef carcasses; and 3) the relationship between indicator bacteria (E. coli and Enterobacteriaceae) counts and visual fecal contamination of beef carcasses. According to the 41 retrieved papers the following conclusions were drawn. A decrease of the indicator bacteria counts was recorded after sequential decontamination treatments, such as pasteurization and hot water washing. Slaughterhouse characteristics influenced bacterial load of beef carcasses, although it was difficult to assess which factors (i.e., slaughterhouse throughput, design of the plant, surveillance system in place) had the greatest effect. Finally, carcasses from fecal contaminated animals had higher bacterial loads than those from clean animals. Therefore, the development of a visual classification system of the level of dirtiness of carcasses and the application of effective treatments on the carcasses classified as dirty along the slaughterline can lead to a contamination level for these carcasses comparable to or lower than that of originally clean ones at the end of the slaughterline.

`Visual inspection of produce fields for signs of fecal contamination and animal intrusion prior to harvest is a currently prescribed method for reducing risk of foodborne illnesses. This report details the continuing development of a fluorescence imaging system to augment human visual inspection. Previous research established that chlorophyll and related compounds, commonly found in feces and injury sites on plants, can be detected using fluorescent responses to UV excitation. In this study, a time-resolved multispectral imaging system was modified to allow hyperspectral line-scan imaging of fluorescent responses to 355-nm pulsed-laser excitation. Addition of a spectral adapter allowed acquisition of line images representing one spatial dimension with full spectral information for each spatial pixel location. Full-object images can be created by concatenating sequential line-scan images. For line-scan imaging, full-object illumination is inefficient. To better fit the illumination to the line-scan imaging field, laser line-expansion was achieved using a Powell lens. Illumination efficiency was 28.5% using the Powell lens compared to 3.0% using simple optical expansion. To test the modified system, spinach was inoculated with 1:2, 1:10, 1:100, and 1:200 dilutions of bovine manure. Using detection based on visual observation, a 100% detection rate was found for all dilutions. Automated detection rates using estimates of fluorescent decay were 100%, 100%, 100%, and 82 % for the 1:2, 1:10, 1:100, and 1:200 dilutions, respectively. These results suggest this technology has potential for the development of a commercial system for pre-harvest detection of fecal contamination and signs of animal intrusion.

Abstract: Simple, automated methods are required for rapid detection of wastewater contamination in urban recreational water. The activity of the enzyme b-D-galactosidase (GAL) can rapidly (5 LOG10. Furthermore, the GAL-activity per gram feces from birds, sheep and cattle was 2–3 LOG10 lower than the activity from human feces, indicating that high GAL-activity in water may reflect human fecal pollution more than the total fecal pollution. The rapid method can only be used to quantify high levels of human fecal pollution, corresponding to about 0.1 mg human feces/liter (or 103 E. coli/100 mL), since below this limit GAL-activity from non-fecal environmental sources may interfere.

International audience; Occurrences of antibiotic-resistant Escherichia coli in two springs of a karstic system (NW France) providing drinking water were determined to study the role of aquifers in the dissemination of the resistance genes. Water samples were collected during wet and dry periods and after a heavy rainfall event to investigate E. coli density, antibiotic resistance patterns, and occurrences of class 1, 2, and 3 integrons. By observing patterns of the resistant isolates (i.e. number and type of resistances) and their occurrences, we were able to define two resistant subpopulations, introduced in the aquifer via surface water: (1) R1-2, characterized by one or two resistance(s), essentially to chloramphenicol and/or tetracycline (96.5%), was always found during the heavy rainfall event; (2) R3-10, characterized by three or more resistances, mostly resistant to tetracycline (94.1%) and beta-lactams (86%), was found transiently. Class 1 and 2 integrons were detected, mostly in the R3-10 subpopulation for class 1 integrons. The characteristics of these two subpopulations strongly suggest that the contamination originates from pasture runoff for the R1-2 subpopulation and from wastewater treatment plant effluents for the R3-10 subpopulation. These two subpopulations of E. coli could be used as biological indicators to determine the origin of groundwater contamination.

This report presents the development of a pre-harvest method and post-harvest technique used to image produce in order to improve food safety and quality. The pre-harvest method includes developing a time-resolved, hyperspectral line scan laser-induced fluorescence imaging system to detect fecal contamination in crop fields. Experimental studies show the effective homogenization of the system's laser and the ability of this system to accurately detect an artificially contaminated apple. The post-harvest technique involves orienting tomatoes to facilitate machine vision for non-destructive imaging. This expands on previous studies that indicate the ability to orient quasi-round fruit their inertial properties. Orientation test were conducted using three tomato cultivars which represented the extremes in potential shapes of tomatoes. Results support the theoretical finding that elongated and squat objects are relatively easy to orient, and round objects are difficult to orient. Both devices are commercially-viable and address the current needs of the food industry.

A two-year ecological study in the Oliveri-Tindari (Messina) brackish water ponds evaluated the quantitative and qualitative distribution of Vibrio species in relation to the physicochemical conditions of the ponds. The evaluation of 204 samples shows a general relationship between the halophilic vibrios and water temperature. In particular this relationship is stronger for the Vibrio fraction able to grow at 37°C, i. e., those species related to human infections. On the other hand, the halophilic vibrios show no relation with the index of fecal contamination. The ecology of V. parahaemolyticus is discussed in relation to the salinity of Marinello pond. The distribution of V. cholerae non O1, isolated from all ponds, shows that this microorganism can be considered as autochthonous in coastal environments.

Introduction : "The occurrence of pathogenic microorganisms in seawater or in shellfish could exist anytime sewage from human or animal origin would be discharged to the coast" (Metcalf, 1982). According to the diseases occurring in the human population or in animals, pathogens might be present in recreational waters or in shellfish. Thus, the presence of human enteric viruses (norovirus, astrovirus, rotavirus, hepatitisAvirus (HAV)) and pathogenic bacteria (Salmonella, Listeria monocytogenes, Shiga-toxin-producing Escherichia coli (STEC), Vibrio cholerae, Vibrio parahaemolyticus, etc.) has been reported in coastal areas for a long time (Colwell, 1978; Metcalf, 1978; Melnick et al., 1979; Grimes, 1991; Bosch et al., 2001;Kong et al., 2002). These microorganisms have been implicated in gastrointestinal and respiratory illnesses and other infections (skin, eyes, etc.), (Griffin et al., 2003). Using risk-assessment models for viruses, maximum risks were estimated to be 1.3 infections per 100 swimmers (Colwell et al., 1996). To evaluate the risk due to the presence of these pathogens in the environment, certain criteria have to be determined. Among them, the infectious dosewould be of a greatest importance (Table 14.1). Even if the infectious dose vary with the strains, the age of the patient, or other parameters, some pathogens are highly dangerous for men even at low concentrations (HAV, E. coli O157:H7, V. cholerae), whereas others have to be ingested in high concentrations to be harmful (V. parahaemolyticus) or are highly infectious but not very dangerous (norovirus). Thus, for some pathogens, a low contamination in seafood, for example, is not acceptable, based on risk-assessment models (Colwell et al., 1996). Among bacteria, the Vibrio family plays an important role in infections, waterborne or seafood diseases, especially in countries surrounded by warm marine waters. Toxigenic V. cholerae O1 and O139 are the causative agents of cholera in developing countries, whereas, non-O1 and non-O139 strains may also be pathogenic but are rarely associated with epidemics (Anonymous, 2002). V. parahaemolyticus has been recognized as a major cause of foodborne gastroenteritis in Japan and Eastern countries and has been linked to seafood consumption. In the United States, more than 700 cases of illness due to V. parahaemolyticus and associated to raw oyster consumption were reported between 1997 and 1998. However, only a few outbreaks have been reported in Australia (1990-1992) and France (2001). The difference between the countries could be attributed to the ecology of this bacteria that requires elevated temperature to grow (>18?C). Vibrio vulnificus associated with primary septicemia, has also been detected in a variety of raw seafood products in Korea and Japan (Anonymous, 2002). Concerning enteric bacteria discharged into marine waters, bacterial control of seafood lowered the risk of Salmonella outbreaks. The occurrence of other bacteria such as Campylobacter jejuni and L. monocytogenes has rarely been reported in shellfish (Feldhusen, 2000; Montfort et al., 1998; Dupray et al., 1999; Federighi et al., 1998). However, L. monocytogenes have been implicated with smoked fish consumption (Rocourt et al., 2000). Human enteric viruses are involved in more than 30% of the outbreaks linked to oyster consumption. Despite the variety of viruses that can communicate shellfish only, HAV and norovirus have been epidemiologically linked to disease (Lees, 2000; Le Guyader et al., 2003). The objective of this review is to gather information on the input and behavior of the main enteric microorganisms-Vibrio family excepted-via the water route and the factors able to affect the seawater and seafood quality.

Using the membrane filtration technique to count total coliform (TC), fecal coliform (FC) and fecal streptococci (FS), the microbiological water quality of the Mfoundi River and four of its representative tributaries at Yaounde, Cameroon, was assessed for human use and contact. Sampling was conducted so as to examine the potential origin of fecal contamination and how rainfall affects the measured concentrations of indicators organisms. Our results revealed that waters were not safe for human use or primary contact according to the standards for water quality established by the Word Health Organization (WHO). Indeed, these waters exhibited high concentrations of TC (Mean +/- SD = 5.6 x 10(8) +/- 2.5 x 10(6) CFU/100 ml), FC (Mean +/- SD = 6.8 x 10(5) +/- 2.4 x 10(3) CFU/100 ml) and FS (Mean +/- SD = 7.3 x 10(5) +/- 2.1 x 10(3) CFU/100 ml) that varied with the sampling sites and points. FC/FS ratio suggested that this contamination was more from warm-blooded animals than humans and correlation analysis points to the role of rainfall as a contributing factor, which enhanced the bacterial numbers detected. We conclude that there is a great potential risk of infection for users of waters from the Mfoundi River and its tributaries at Yaounde.

A study on the distribution patterns of enteropathogenic bacteria polluting the shoreline in Natal, Rio Grande do Norte, Brazil, was carried out based on 72 samples obtained from three storm sewers and adjoining beach locations, Praia do Meio (PM), Areia Preta (AP) and Ponta Negra (PN). From each location, 12 water samples were taken and analyzed for fecal coliforms (FC) and Escherichia coli. In AP, two (16.7%) of the seawater samples and five (41.7%) of the storm sewer samples yielded values above 1.1 × 10(7) FC/100 ml, whereas only one (8.3%) of the samples from PM reached this level. There was no correlation (p > 0.05) between rainfall indeces and FC values. A total of 64 E. coli isolates were obtained: 37 from the storm sewer samples and 27 from the seawater samples. Of these isolates, four (O143, two O112ac, and O124) were enteroinvasive and two (O111 and O125) were enteropathogenic. Resistance to antibiotics and to heavy metals was also analyzed. Almost 36% of the E. coli strains isolated were resistant to more than one antibiotic. All strains were resistant to zinc and copper at the highest concentration tested (250 µg/ml), and several (23.4%) were resistant to mercury at 50 µg/ml. Our results agreed with previous reports that antibiotic resistance is commonly associated with heavy-metal resistance in pathogens.

The first case of human cryptosporidiosis was reported in Korea in 1995; however, an outbreak of Cryptosporidium has not been reported in Korea until now. This paper describes the first outbreak of cryptosporidiosis in Korea. On May 24, 2012, a local public health center filed a report on 126 residents with gastrointestinal symptoms in an old apartment complex in Seoul. Epidemiological investigations were implemented on 125 of the 126 patients. The patients were reported continuously over a period of 22 days. Diarrhea was the most common clinical symptom, and lasted for 5 days on average. The tap water was the only common exposure of the patients. During the environmental investigation it was discovered that the water and septic tanks were situated closely and that the waste water pipes were corroded where they passed over the water pipes. Cryptosporidium parvum was detected in 3 of the 7 stool specimens by PCR-RFLP. A number of Cryptosporidium oocysts were also detected in the water specimens from the water tank. In conclusion, Cryptosporidium parvum was the key causal pathogen of this outbreak. It is presumed that the tap water was contaminated by a sewage leak from the aged pipelines.