Amandine Laffite

Accumulation of Clinically Relevant Antibiotic-Resistance Genes, Bacterial Load, and Metals in Freshwater Lake Sediments in Central Europe

Wastewater treatment plants (WWTP) receive the effluents from various sources (communities, industrial, and hospital effluents) and are recognized as reservoir for antibiotic-resistance genes (ARGs) that are associated with clinical pathogens. The aquatic environment is considered a hot-spot for horizontal gene transfer, and lake sediments offer the opportunity for reconstructing the pollution history and evaluating the impacts. In this context, variation with depth and time of the total bacterial load, the abundance of faecal indicator bacteria (FIB; E. coli and Enterococcus spp. (ENT)), Pseudomonas spp., and ARGs (blaTEM, blaSHV, blaCTX-M, blaNDM, and aadA) were quantified in sediment profiles of different parts of Lake Geneva using quantitative PCR. The abundance of bacterial marker genes was identified in sediments contaminated by WWTP following eutrophication of the lake. Additionally, ARGs, including the extended-spectrum ß-lactam- and aminoglycoside-resistance genes, were identified in the surface sediments. The ARG and FIB abundance strongly correlated (r ≥ 0.403, p < 0.05, n = 34) with organic matter and metal concentrations in the sediments, indicating a common and contemporary source of contamination. The contamination of sediments by untreated or partially treated effluent water can affect the quality of ecosystem. Therefore, the reduction of contaminants from the source is recommended for further improvement of water quality.

Hospital Effluents Are One of Several Sources of Metal, Antibiotic Resistance Genes, and Bacterial Markers Disseminated in Sub-Saharan Urban Rivers

Data concerning the occurrence of emerging biological contaminants such as antibiotic resistance genes (ARGs) and fecal indicator bacteria (FIB) in aquatic environments in Sub-Saharan African countries is limited. On the other hand, antibiotic resistance remains a worldwide problem which may pose serious potential risks to human and animal health. Consequently, there is a growing number of reports concerning the prevalence and dissemination of these contaminants into various environmental compartments. Sediments provide the opportunity to reconstruct the pollution history and evaluate impacts so this study investigates the abundance and distribution of toxic metals, FIB, and ARGs released from hospital effluent wastewaters and their presence in river sediments receiving systems. ARGs (blaTEM, blaCTX-M, blaSHV, and aadA), total bacterial load, and selected bacterial species FIB [Escherichia coli, Enterococcus (ENT)] and species (Psd) were quantified by targeting species specific genes using quantitative PCR (qPCR) in total DNA extracted from the sediments recovered from 4 hospital outlet pipes (HOP) and their river receiving systems in the City of Kinshasa in the Democratic Republic of the Congo. The results highlight the great concentration of toxic metals in HOP, reaching the values (in mg kg−1) of 47.9 (Cr), 213.6 (Cu), 1434.4 (Zn), 2.6 (Cd), 281.5 (Pb), and 13.6 (Hg). The results also highlight the highest (P < 0.05) values of 16S rRNA, FIB, and ARGs copy numbers in all sampling sites including upstream (control site), discharge point, and downstream of receiving rivers, indicating that the hospital effluent water is not an exclusive source of the biological contaminants entering the urban rivers. Significant correlation were observed between (i) all analyzed ARGs and total bacterial load (16S rRNA) 0.51 to 0.72 (p < 0.001, n = 65); (ii) ARGs (except blaTEM) and FIB and Psd 0.57 < r < 0.82 (p < 0.001, n = 65); and (iii) ARGs (except blaTEM) and toxic metals (Cd, Cr, Cu, and Zn) 0.44 to 0.72, (p < 0.001, n = 65). These findings demonstrate that several sources including hospital and urban wastewaters contribute to the spread of toxic metals and biological emerging contaminants in aquatic ecosystems.

Leachates draining from controlled municipal solid waste landfill: Detailed geochemical characterization and toxicity tests

Management of municipal solid wastes in many countries consists of waste disposal into landfill without treatment or selective collection of solid waste fractions including plastics, paper, glass, metals, electronic waste, and organic fraction leading to the unsolved problem of contamination of numerous ecosystems such as air, soil, surface, and ground water. Knowledge of leachate composition is critical in risk assessment of long-term impact of landfills on human health and the environment as well as for prevention of negative outcomes. The research presented in this paper investigates the seasonal variation of draining leachate composition and resulting toxicity as well as the contamination status of soil/sediment from lagoon basins receiving leachates from landfill in Mpasa, a suburb of Kinshasa in the Democratic Republic of the Congo. Samples were collected during the dry and rainy seasons and analyzed for pH, electrical conductivity, dissolved oxygen, soluble ions, toxic metals, and were then subjected to toxicity tests. Results highlight the significant seasonal difference in leachate physicochemical composition. Affected soil/sediment showed higher values for toxic metals than leachates, indicating the possibility of using lagoon system for the purification of landfill leachates, especially for organic matter and heavy metal sedimentation. However, the ecotoxicity tests demonstrated that leachates are still a significant source of toxicity for terrestrial and benthic organisms. Therefore, landfill leachates should not be discarded into the environment (soil or surface water) without prior treatment. Interest in the use of macrophytes in lagoon system is growing and toxic metal retention in lagoon basin receiving systems needs to be fully investigated in the future. This study presents useful tools for evaluating landfill leachate quality and risk in lagoon systems which can be applied to similar environmental compartments.

Occurrence of Antibiotic Resistance Genes and Bacterial Markers in a Tropical River Receiving Hospital and Urban Wastewaters

The occurrence of emerging biological contaminants including antibiotic resistance genes (ARGs) and Faecal Indicator Bacteria (FIB) is still little investigated in developing countries under tropical conditions. In this study, the total bacterial load, the abundance of FIB (E. coli and Enterococcus spp. (ENT)), Pseudomonas spp. and ARGs (blaTEM, blaCTX-M, blaSHV, blaNDM and aadA) were quantified using quantitative PCR in the total DNA extracted from the sediments recovered from hospital outlet pipes (HOP) and the Cauvery River Basin (CRB), Tiruchirappalli, Tamil Nadu, India. The abundance of bacterial marker genes were 120, 104 and 89 fold higher for the E. coli, Enterococcus spp. and Pseudomonas spp., respectively at HOP when compared with CRB. The ARGs aadA and blaTEM were most frequently detected in higher concentration than other ARGs at all the sampling sites. The ARGs blaSHV and blaNDM were identified in CRB sediments contaminated by hospital and urban wastewaters. The ARGs abundance strongly correlated (r ≥ 0.36, p < 0.05, n = 45) with total bacterial load and E. coli in the sediments, indicating a common origin and extant source of contamination. Tropical aquatic ecosystems receiving wastewaters can act as reservoir of ARGs, which could potentially be transferred to susceptible bacterial pathogens at these sites.

The impact of hospital and urban wastewaters on the bacteriological contamination of the water resources in Kinshasa, Democratic Republic of Congo.

ABSTRACT Although the United Nations General Assembly recognized in 2010 the right to safe and clean drinking water and sanitation as a human right that is essential to the full enjoyment of life and all other human rights, the contamination of water supplies with faecal pathogens is still a major and unsolved problem in many parts of the world. In this study, faecal indicator bacteria (FIB), including Escherichia coli (E. coli) and Enterococcus (ENT), were quantified over the period of June/July 2014 and June/July 2015 to assess the quality of hospital effluents (n = 3: H1, H2 and H3) and of rivers receiving wastewaters from the city of Kinshasa, Democratic Republic of Congo. The water and sediment samples from the river-receiving systems were collected in, upstream and downstream of the hospital outlet pipe (HOP) discharge. The analysis of E. coli and ENT in water and sediment suspension was performed using the cultural membrane filter method. The FIB characterization was performed for general E. coli, Enterococcus faecalis(E. faecalis) and human-specific Bacteroides by PCR using specific primers. The results revealed very high FIB concentration in the hospital effluent waters, with E. coli reaching the values of 4.2 × 105, 16.1 × 105 and 5.9 × 105 CFU 100 mL−1, for the hospital effluents from H1, H2, and H3, respectively; and Enterococcus reaching the values of 2.3 × 104, 10.9 × 104 and 4.1 × 104 CFU 100 mL−1, respectively. Interestingly, the FIB levels in the water and sediment samples from river-receiving systems are spatially and temporally highly variable and present in some samples with higher values than the hospital effluents. The PCR assays for human-specific Bacteroides HF183/HF134 further indicate that more than 98% of bacteria were from human origin. The results of this research therefore confirm the hypothesis of our previous studies, indicating that in developing countries (e.g., Democratic Republic of Congo and South India), the hospital effluent waters can be a significant source of the deterioration of the bacteriological quality for urban rivers. The approach used in this investigation can be further used to decipher the pollution of water resources by human faecal contamination. The results of this research will help to better understand the microbiological pollution problems in river-receiving systems and will guide municipality decisions on improving the urban water quality.

Microbiological quality of water in a city with persistent and recurrent waterborne diseases under tropical sub-rural conditions: The case of Kikwit City, Democratic Republic of the Congo

The availability of safe drinking water in sub-Saharan countries remains a major challenge because poor sanitation has been the cause of various outbreaks of waterborne disease due to the poor microbiological quality of water used for domestic purposes. The faecal indicator bacteria (FIB) used in the present study included Escherichia coli (E. coli) and Enterococcus (ENT). FIB and aerobic mesophilic bacteria (AMB) were quantified during July 2015 (dry season) and November 2015 (rainy season) in order to assess the quality of drinking water from wells (n=3; P1-P3), and two rivers, the River Lukemi (RLK, n=3) and River Luini (RLN, n=2) in the city of Kikwit, which is located in the province of Kwilu in the Democratic Republic of the Congo. Kikwit is well known for its outbreaks of persistent and recurrent waterborne diseases including Entamoeba, Shigella, typhoid fever, cholera, and Ebola Viral Hemorrhagic Fever. Consequently, E. coli, ENT, and AMB were quantified in water samples according to the standard international methods for water quality determination using the membrane filtration method. The FIB characterization was performed for human-specific Bacteroides by PCR using specific primers. The results obtained revealed high FIB concentrations in river samples collected during both seasons. For example, E. coli respectively reached 4.3×104 and 9.2×104 CFU 100mL-1 in the dry season and the wet season. ENT reached 5.3×103 CFU 100mL-1 during the dry season and 9.8×103 CFU 100mL-1 in the wet season. The pollution was significantly worse in the wet season compared to the dry season. Surprisingly, no faecal contamination was observed in well water samples collected in the dry season while E. coli and ENT were detected in all wells in the wet season with values of 6, 7, and 11CFUmL-1 for E. coli in wells P1-P3, respectively and 3, 5, 9 CFU mL-1for ENT in the same wells. Interestingly, the PCR assays for human-specific Bacteroides HF183/HF134 indicated that 97-100% captured in all analyses of isolated FIB were of human origin. The results indicate that contamination of E. coli, ENT, and AMB in the studied water resources increases during the wet season. This study improves understanding of the microbiological pollution of rivers and wells under tropical conditions and will guide future municipal/local government decisions on improving water quality in this region which is characterised by persistent and recurrent waterborne diseases. Although the epidemiology can be geographically localised, the effects of cross border transmission can be global. Therefore, the research results presented in this article form recommendations to municipalities/local authorities and the approach and procedures can be carried out in a similar environment.

High levels of faecal contamination in drinking groundwater and recreational water due to poor sanitation, in the sub-rural neighbourhoods of Kinshasa, Democratic Republic of the Congo

In many urban and peri-urban areas of developing countries, shallow wells and untreated water from urban rivers are used for domestic purposes, including drinking water supply, population bathing and irrigation for urban agriculture. The evaluation and monitoring of water quality are therefore necessary for preventing potential human risk associated with the exposure to contaminated water. In this study, physicochemical and bacteriological parameters were assessed in an urban river (named Kokolo Canal/Jerusalem River) draining the municipality of Lingwala (City of Kinshasa, Democratic Republic of the Congo) and in two shallow wells used as drinking water supplies, during the wet and dry seasons in order to estimate the seasonal variation of contamination. The faecal indicator bacteria (FIB) isolated strains (Escherichia coli (E. coli) and Enterococcus (ENT)) from water and surface sediment, were characterized for human-specific bacteroides by molecular approach. The results revealed very high faecal contamination of water from the shallow wells, and of water and sediments from the river, during both wet and dry seasons. During the wet season, E. coli reached the values of 18.6 × 105 and 4.9 × 105 CFU 100 mL-1 in Kokolo Canal and shallow wells, respectively; and Enterococcus reached the values of 7.4 × 104 and 2.7 × 104 CFU 100 mL-1. Strong mutually positive correlation was observed between E. coli and ENT, with the range of R-value being 0.93 < r < 0.97 (p-value < 0.001, n = 15). The PCR assays for human-specific Bacteroides indicated that more than 98% of 500 isolated FIB strains were of human origin, pointing out the effect of poor household sanitation practices on surface water but also on groundwater contamination. The water samples from the shallow wells and Kokolo Canal were highly polluted with faecal matter in both seasons. However, the pollution level was significantly higher during the wet season compared to the dry season. Physicochemical analysis revealed also very high water electrical conductivity, with values much higher than the recommended limits of the World Health Organization guideline for drinking water. These results highlight the potential human health risk associated with the exposure to water contamination from shallow wells and Kokolo Canal, due to the very high level of human FIB. Rapid, unplanned and uncontrolled population growth in the city of Kinshasa is increasing considerably the water demand, whereas there is a dramatic lack of appropriate sanitation and wastewater facilities, as well as of faecal sludge (and solid waste) management and treatment. The lack of hygiene and the practice of open defecation is leading to the degradation of water quality, consequently the persistence of waterborne diseases in the neighbourhoods of sub-rural municipalities, and there is a growing threat to the sustainability to water resources and water quality. The results of this study should encourage municipality policy and strategy on increasing the access to safely managed sanitation services; in order to better protect surface water and groundwater sources, and limit the proliferation of epidemics touching regularly the city.