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The increase in industrial production of multi-walled carbon nanotubes (MWCNTs) raises concerns about their potential adverse effects associated to environmental releases, especially in aquatic environments where they are likely to accumulate. This study focuses on the environmental impact of MWCNTs, specifically on a benthic freshwater diatom (Nitzschia linearis), which plays a major role in the primary production of water bodies. The obtained results indicate that exposure to MWCNTs in the presence of natural organic matter (NOM) inhibits diatom's growth in a dose-dependent manner after 72 h of exposure. Interestingly, the photosystem II quantum yield (PSII QY ) in diatoms remains unaffected even after exposure to MWCNTs at 10 mg/L. After 48 h of exposure, MWCNTs are found to bind preferentially to extracellular polymeric substances (EPS) produced by diatoms, which could decrease their toxicity by limiting their interaction with this organism. However, measurement of genotoxicity and teratogenicity in diatoms exposed to MWCNTs revealed that the exposure to MWCNTs increased the occurrence of cells with micronuclei and abnormal frustules. Microscopy analyses including two-photon excitation microscopy (TPEM) revealed the internalization of MWCNTs. Investigations of the diatom's frustule structure using Scanning electron microscopy (SEM) indicated that the presence of pore structures constitutes a pathway allowing MWCNTs uptake. The presence in the diatom's cytoplasm of MWCNTs might possibly induce disturbances of the cellular components, leading to the observed genotoxic and teratogenic effects. In view of previous studies, this work underscores the need for further studies on the interaction between nanomaterials and different diatom species, given the species-specific nature of the interactions., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Antibiotic resistance (AR) is one of the major health threats of our time. The presence of antibiotics in the environment and their continuous release from sewage treatment plants, chemical manufacturing plants and animal husbandry, agriculture and aquaculture, result in constant selection pressure on microbial organisms. This presence leads to the emergence, mobilization, horizontal gene transfer and a selection of antibiotic resistance genes, resistant bacteria and mobile genetic elements. Under these circumstances, aquatic wildlife is impacted in all compartments, including freshwater organisms with partially impermeable microbiota. In this narrative review, recent advancements in terms of occurrence of antibiotics and antibiotic resistance genes in sewage treatment plant effluents source compared to freshwater have been examined, occurrence of antibiotic resistance in wildlife, as well as experiments on antibiotic exposure. Based on this current state of knowledge, we propose the hypothesis that freshwater aquatic wildlife may play a crucial role in the dissemination of antibiotic resistance within the environment. Specifically, we suggest that organisms with high bacterial density tissues, which are partially isolated from the external environment, such as fishes and amphibians, could potentially be reservoirs and amplifiers of antibiotic resistance in the environment, potentially favoring the increase of the abundance of antibiotic resistance genes and resistant bacteria. Potential avenues for further research (trophic transfer, innovative exposure experiment) and action (biodiversity eco-engineering) are finally proposed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Due to their properties, graphene-based nanomaterials (GBMs) are triggering a great interest leading to an increase of their global production and use in new applications. As a consequence, their release into the environment is expected to increase in the next years. When considering the current knowledge in the evaluation of GBMs ecotoxic potential, studies aiming to evaluate the hazard associated to these nanomaterials towards marine species and particularly considering potential interactions with other environmental pollutants such as metals are scarce. Here we evaluated the embryotoxic potential of GBMs, which include graphene oxide (GO) and its reduced form (rGO), both individually and in combination with copper (Cu) as a referent toxicant, towards early life stages of the Pacific oyster through the use of a standardized method (NF ISO 17244). We found that following exposure to Cu, dose-dependent decrease in the proportion of normal larvae was recorded with an Effective Concentration leading to the occurrence of 50% of abnormal larvae (EC 50 ) of 13.85 ± 1.21 μg/L. Interestingly, the presence of GO at a non-toxic dose of 0.1 mg/L decreased the Cu EC 50 to 12.04 ± 0.85 μg/L while it increased to 15.91 ± 1.57 μg/L in presence of rGO. Based on the measurement of copper adsorption, the obtained results suggest that GO enhances Cu bioavailability, potentially modifying its toxic pathways, while rGO mitigates Cu toxicity by decreasing its bioavailability. This research underscores the need to characterize the risk associated to GBMs interactions with other aquatic contaminants and supports the adoption of a safer-by-design strategy using rGO in marine environments. This would contribute to minimize the potential adverse effects on aquatic species and to reduce the risk for economic activities associated to coastal environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Despite the fast-growing use and production of graphene-based nanomaterials (GBMs), data concerning their effects on freshwater benthic macroinvertebrates are scarce. This study aims to investigate the effects of graphene oxide (GO) on the midge Chironomus riparius . Mortality, growth inhibition, development delay and teratogenicity, assessed using mentum deformity analysis, were investigated after a 7-day static exposure of the first instar larvae under controlled conditions. The collected data indicated that the survival rate was not impacted by GO, whereas chronic toxicity following a dose-dependent response occurred. Larval growth was affected, leading to a significant reduction in larval length (from 4.4 to 10.1%) in individuals reaching the fourth instar at any of the tested concentrations (from 0.1 to 100 mg/L). However, exposure to GO is not associated with an increased occurrence of mouthpart deformities or seriousness in larvae. These results highlight the suitability of monitoring the larval development of C. riparius as a sensitive marker of GO toxicity. The potential ecological consequences of larval size decrease need to be considered for a complete characterization of the GO-related environmental risk.

Due to their various properties as polymeric materials, plastics have been produced, used and ultimately discharged into the environment. Although some studies have shown their negative impacts on the marine environment, the effects of plastics on freshwater organisms are still poorly studied, while they could be widely in contact with this pollution. The current work aimed to better elucidate the impact and the toxicity mechanisms of two kinds of commercial functionalized nanoplastics, i.e., carboxylated polystyrene microspheres of, respectively, 350 and 50 nm (PS350 and PS50), and heteroaggregated PS50 with humic acid with an apparent size of 350 nm (PSHA), all used at environmental concentrations (0.1 to 100 µg L -1 ). For this purpose, two relevant biological and aquatic models-amphibian larvae, Xenopus laevis , and dipters, Chironomus riparius -were used under normalized exposure conditions. The acute, chronic, and genetic toxicity parameters were examined and discussed with regard to the fundamental characterization in media exposures and, especially, the aggregation state of the nanoplastics. The size of PS350 and PSHA remained similar in the Xenopus and Chironomus exposure media. Inversely, PS50 aggregated in both exposition media and finally appeared to be micrometric during the exposition tests. Interestingly, this work highlighted that PS350 has no significant effect on the tested species, while PS50 is the most prone to alter the growth of Xenopus but not of Chironomus. Finally, PSHA induced a significant genotoxicity in Xenopus .

The interest for graphene-based nanomaterials (GBMs) is growing worldwide as their properties allow the development of new innovative applications. In parallel, concerns are increasing about their potential adverse effects on the environment are increasing. The available data concerning the potential risk associated to exposure of aquatic organisms to these GBMs are still limited and little is known regarding their endocrine disruption potential. In the present study, the endocrine disruption potential of graphene oxide (GO) and reduced graphene oxide (rGO) was assessed using a T3-induced amphibian metamorphosis assay. The results indicated that GBMs potentiate the effects of exogenous T3 with a more marked effect of GO compared to rGO. T3 quantifications in the exposure media indicated adsorption of the hormone on GBMs, increasing its bioavailability for organisms because GBMs are accumulated in the gut and the gills of these amphibians. This study highlights that the tested GBMs do not disrupt the thyroid pathway in amphibians but indicates that adsorption properties of these nanomaterials may increase the bioavailability and the toxicity of other pollutants., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Among the challenges in controlling tuberculosis, a rapid and accurate diagnostic test for the detection of Mycobacterium tuberculosis complex (MTBc) and its resistance to first line therapies is crucial. We evaluated the performance of the Xpert MTB/RIF Ultra assay (Xpert Ultra) for the rapid detection of MTBc and rifampicin resistance (RR) in 1120 pulmonary and 461 extra-pulmonary clinical specimens and compared it with conventional phenotypic techniques. The Xpert Ultra assay detected MTBc in 223 (14.1%) samples with an overall sensitivity and specificity, using culture as the "gold standard", of 91.1% (95% CI, 85.6-95.1) and 94.5% (95% CI, 93.1-95.6), respectively. The sensitivity of the Xpert Ultra test for smear-negative extra-pulmonary specimens was high (87.1%), even higher than with smear-negative pulmonary specimens (81.8%). But this enhanced sensitivity came with a low overall specificity of smear-negative extra-pulmonary specimens (66.7%). For 73 patients, 79/1423 (3.4%) negative mycobacterial culture samples were found to be positive with Xpert Ultra. Clinical data was necessary to correctly interpret potential false-positive results, especially trace-positive results. Sensitivity of the Xpert Ultra to detect RR compared to drug susceptibility testing was 100% (95% CI, 29.2-100) and specificity was 99.2% (95% CI, 95.8-100). We concluded that the Xpert Ultra test is able to provide a reliable TB diagnosis within a significantly shorter turnaround time than culture. This is especially true for paucibacillary samples such as smear-negative pulmonary specimens and extra-pulmonary specimens., Competing Interests: No authors have competing interests.

Graphene-based nanomaterials (GBMs), such as graphene oxide (GO) and reduced graphene oxide (rGO), possess unique properties triggering high expectations for the development of new technological applications and are forecasted to be produced at industrial-scale. This raises the question of potential adverse outcomes on living organisms and especially toward microorganisms constituting the basis of the trophic chain in ecosystems. However, investigations on GBMs toxicity were performed on various microorganisms using single species that are helpful to determine toxicity mechanisms but fail to predict the consequences of the observed effects at a larger organization scale. Thus, this study focuses on the ecotoxicological assessment of GO and rGO toward a biofilm composed of the diatom Nitzschia palea associated to a bacterial consortium. After 48 and 144 h of exposure to these GBMs at 0, 0.1, 1, and 10 mg.L -1 , their effects on the diatom physiology, the structure, and the metabolism of bacterial communities were measured through the use of flow cytometry, 16S amplicon sequencing, and Biolog ecoplates, respectively. The exposure to both of these GBMs stimulated the diatom growth. Besides, GO exerted strong bacterial growth inhibition as from 1 mg.L -1 , influenced the taxonomic composition of diatom-associated bacterial consortium, and increased transiently the bacterial activity related to carbon cycling, with weak toxicity toward the diatom. On the contrary, rGO was shown to exert a weaker toxicity toward the bacterial consortium, whereas it influenced more strongly the diatom physiology. When compared to the results from the literature using single species tests, our study suggests that diatoms benefited from diatom-bacteria interactions and that the biofilm was able to maintain or recover its carbon-related metabolic activities when exposed to GBMs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Evariste, Braylé, Mouchet, Silvestre, Gauthier, Flahaut, Pinelli and Barret.)

The environmental fate and behavior of nanoplastics (NPs) and their toxicity against aquatic organisms are under current investigation. In this work, relevant physicochemical characterizations were provided to analyze the ecotoxicological risk of NPs in the aquatic compartment. For this purpose, heteroaggregates of 50 nm polystyrene nanospheres and natural organic matter were prepared and characterized. The kinetic of aggregation was assimilated to a reaction-limited colloid aggregation mode and led to the formation of heteroaggregates in the range of 100-500 nm. Toxicities of these heteroaggregates and polystyrene nanospheres (50 and 350 nm) were assessed for a large range of concentrations using four benthic and one planktonic algal species, in regards to particle states in the media. Heteroaggregates and nanospheres were shown to be stable in the exposure media during the ecotoxity tests. The algal species exhibited very low sensitivity (growth and photosynthetic activity), with the noteworthy exception of the planktonic alga, whose growth increased by more than 150% with the heteroaggregates at 1 µg L -1 . Despite the lack of a strong direct effect of the NPs, they may still impair the functioning of aquatic ecosystems by destabilizing the competitive interactions between species. Moreover, further work should assess the toxicity of NPs associated with other substances (adsorbed pollutants or additives) that could enhance the NP effects.

Introduction: Belgium is a country with low incidence of tuberculosis (TB) and a very low number of TB cases in children. Children in contact with an adult smear-positive TB case are at high risk of transmission. Early diagnosis is important as young children have a significant predisposition of developing TB disease. In this paper, we describe two outbreaks after exposure to, respectively, two teachers with smear-positive pulmonary TB: one in a primary school, a nursery teacher, and another in a private language school. Methods: An exposure investigation was carried out in both index cases household and school, according to the stone-in-the-pond principle. The tuberculin skin test (TST) was used a screening tool. The time elapsed between TB diagnosis in the index case and contact investigation was, respectively, 1 and 3 weeks. If this initial test was negative, it was repeated after a "window period" of ≥8 weeks. Results: Index cases showed a transmission rate of, respectively, 13 and 40% in their classes at school, defined as casual contacts. The proximity of contact increased the risk of infection. TB disease was observed in, respectively, 4 and 11% of all the casual contacts; all of them were children younger than 5 years old. TB-infected and children with active TB disease had good compliance with recommended treatment. Uptake of chemoprophylaxis during the "window period" was poor, respectively, only 32-42%, in children under 5 years with an initially negative TST. Discussion: The World Health Organization recommends to screen all young children (<5 years old) who have close contact with a person affected by pulmonary TB and to initiate Latent tuberculosis infection treatment even before infection can be demonstrated, after ruling out active TB disease. Despite this knowledge, a small percentage of the children younger than 5 years with no proof of infection was treated with the proposed chemoprophylactic treatment, in both cases. Conclusion: This exposure investigation of two teachers detects high transmission among family contacts and school casual contacts. Recommendations for chemoprophylactic treatment in children <5 years showed low compliance, reflecting the difficulty of communication to staff, parents, and children in a school outbreak. It is essential to develop a new approach for this vulnerable group of patients. This approach could be improved, applied, and evaluated by National TB Control Programs, involving public and private health services. Public health authorities play a role in raising public awareness about the risks of TB for young children., (Copyright © 2020 Debulpaep, Dreesman, Dirix, Toppet, Wanlin, Geysens, Arrazola de Oñate, Fauville, Mascart, Levy and Mouchet.)

Background: Improved diagnostic tests are needed for the early identification of Mycobacterium tuberculosis- infected young children exposed to an active TB (aTB) index case. We aimed to compare the diagnostic accuracy of new blood-based tests to that of the tuberculin skin test (TST) for the identification of all infected children and for a potential differentiation between aTB and latent TB infection (LTBI). Methods: 144 children exposed to a patient with aTB were included, and those who met all inclusion criteria (130/144) were classified in three groups based on results from classical investigations: non-infected (NI: n = 69, 53%, median age 10 months), LTBI ( n = 28, 22%, median age 96 months), aTB disease ( n = 33, 25%, median age 24 months). The first whole blood assay consisted of a 7-days in vitro stimulation of blood with four different mycobacterial antigens (40 μl/condition), followed by flow cytometric measurement of the proportions of blast cells appearing among lymphocytes as a result of their specific activation. Thresholds of positivity were determined by Receiver Operating Characteristic (ROC) curve analysis (results of NI children vs. children with LTBI/aTB) in order to identify infected children in a first stage. Other cut-offs were determined to discriminate subgroups of infected children in a second step (results from children with aTB/LTBI). Analysis of blood monocytes and dendritic cell subsets was performed on 100 μl of blood for 25 of these children as a second test in a pilot study. Results: Combining the results of the blast-induced CD3 + T lymphocytes by Heparin-Binding Haemagglutinin and by Culture Filtrate Protein-10 identified all but one infected children (sensitivity 98.2% and specificity 86.9%, compared to 93.4 and 100% for the TST). Further identification among infected children of those with aTB was best achieved by the results of blast-induced CD8 + T lymphocytes by purified protein derivative (sensitivity for localized aTB: 61.9%, specificity 96.3%), whereas high proportions of blood type 2 myeloid dendritic cells (mDC) were a hallmark of LTBI. Conclusions: New blood-based tests requiring a very small volume allow the accurate identification of M. tuberculosis -infected young children among exposed children and are promising to guide the clinical classification of children with aTB or LTBI.

Introduction: Interferon Gamma Release Assay (IGRA) has proven to be a useful test to evaluate the immune response to Mycobacterium tuberculosis antigens in children over the age of 5 years as an alternative to tuberculin skin testing (TST). Much less is known about its performance in younger children, who are at higher risk for developing tuberculosis (TB) disease after exposure. We aimed to evaluate the accuracy of using IGRA in TB screening in this population. Methods: Children below the age of 5 years at high risk for TB infection were prospectively enrolled, to compare the performance of TST and the QuantiFERON-TB Gold-In-Tube test (QFT). Children were treated in accordance with the diagnosis made at baseline and followed-up for 12 months. Results: We included a total of 60 children of which 97 blood samples were available for analysis. There was 90.72% agreement between TST and QFT (Kappa test 0.59, moderate agreement). With TST as a reference, the QFT positive predictive value was 0.72 and the negative predictive value 0.93. Discordant results were observed with 6% TST+/QFT- paired tests. When we restricted the comparison of TST and QFT to non-BCG-vaccinated children, the degree of agreement was more substantial (95%, Kappa test 0.75) and the negative predictive value was 0.99. We observed 3% discordant TST-/QFT+ results. All children with active TB disease had concordant positive QFT results, with QFT values above 4.00 IU/ml. Conclusion: In a low TB prevalence country, serial testing of QFT was found to produce a moderate agreement with TST results. False positive QFT results would have been eliminated by using a higher cutoff without misdiagnosing the children with TB disease. Some of the false negative QFT results could be explained by false positive TST results on consecutive testing. For now the most prudent approach would be to consider discordant QFT-/TST+ results as false negative QFT results, taking into account the young age of our population and the potential risk for evolution to active TB disease.

Gut microbial communities constitute a compartment of crucial importance in regulation of homeostasis of multiple host physiological functions as well as in resistance towards environmental pollutants. Many chemical contaminants were shown to constitute a major threat for gut bacteria. Changes in gut microbiome could lead to alteration of host health. The access to high-throughput sequencing platforms permitted a great expansion of this discipline in human health while data from ecotoxicological studies are scarce and particularly those related to aquatic pollution. The main purpose of this review is to summarize recent body of literature providing data obtained from microbial community surveys using high-throughput 16S rRNA sequencing technology applied to aquatic ecotoxicity. Effects of pesticides, PCBs, PBDEs, heavy metals, nanoparticles, PPCPs, microplastics and endocrine disruptors on gut microbial communities are presented and discussed. We pointed out difficulties and limits provided by actual methodologies. We also proposed ways to improve understanding of links between changes in gut bacterial communities and host fitness loss, along with further applications for this emerging discipline., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The worldwide increase of graphene family materials raises the question of the potential consequences resulting from their release in the environment and future consequences on ecosystem health, especially in the aquatic environment in which they are likely to accumulate. Thus, there is a need to evaluate the biological and ecological risk but also to find innovative solutions leading to the production of safer materials. This work focuses on the evaluation of functional group-safety relationships regarding to graphene oxide (GO) in vivo genotoxic potential toward X. laevis tadpoles. For this purpose, thermal treatments in H₂ atmosphere were applied to produce reduced graphene oxide (rGOs) with different surface group compositions. Analysis performed indicated that GO induced disturbances in erythrocyte cell cycle leading to accumulation of cells in G0/G1 phase. Significant genotoxicity due to oxidative stress was observed in larvae exposed to low GO concentration (0.1 mg.L - ¹). Reduction of GO at 200 °C and 1000 °C produced a material that was no longer genotoxic at low concentrations. X-ray photoelectron spectroscopy (XPS) analysis indicated that epoxide groups may constitute a good candidate to explain the genotoxic potential of the most oxidized form of the material. Thermal reduction of GO may constitute an appropriate "safer-by-design" strategy for the development of a safer material for environment., Competing Interests: The authors declare no conflict of interest.

Nano-ecotoxicology is an emerging science which aims to assess the environmental effect of nanotechnologies. The development of this particular aspect of ecotoxicology was made necessary in order to evaluate the potential impact of recently produced and used materials: nanoparticles (NPs). Among all the types of NPs, carbon nanoparticles (CNPs) especially draw attention giving the increasing number of applications and integration into consumer products. However the potential impacts of CNPs in the environment remain poorly known. This review aims to point out the critical issues and aspects that will govern the toxicity of CNPs in the environment., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: Arboviral diseases are an important public health concerns. Vector control remains the sole strategy to fight against these diseases. Because of the important limits of methods currently used to assess human exposure to Aedes mosquito bites, much effort is being devoted to develop new indicators. Recent studies have reported that human antibody (Ab) responses to Aedes aegypti Nterm-34kDa salivary peptide represent a promising biomarker tool to evaluate the human-Aedes contact. The present study aims investigate whether such biomarker could be used for assessing the efficacy of vector control against Aedes., Methodology/principal Findings: Specific human IgG response to the Nterm-34kDa peptide was assessed from 102 individuals living in urban area of Saint-Denis at La Reunion Island, Indian Ocean, before and after the implementation of vector control against Aedes mosquitoes. IgG response decreased after 2 weeks (P < 0.0001), and remained low for 4 weeks post-intervention (P = 0.0002). The specific IgG decrease was associated with the decline of Aedes mosquito density, as estimated by entomological parameters and closely correlated to vector control implementation and was not associated with the use of individual protection, daily commuting outside of the house, sex and age. Our findings indicate a probable short-term decrease of human exposure to Aedes bites just after vector control implementation., Conclusion/significance: Results provided in the present study indicate that IgG Ab response to Aedes aegypti Nterm-34kDa salivary peptide could be a relevant short-time indicator for evaluating the efficacy of vector control interventions against Aedes species., Competing Interests: The authors have declared that no competing interests exist.

The aim of the present work is to provide wider information on the toxicity of cerium dioxide nanoparticles (CeO2 NPs) in aquatic environments, by studying the toxicity of two types of CeO2 NPs for four species (diatoms Nitzschia palea, the sediment-dwelling invertebrate Chironomus riparius, and the amphibian larvae Xenopus laevis and Pleurodeles waltl.). The two types of CeO2 NPs have different intrinsic properties: some of them are small citrate-coated spheres (2-5 nm), and the others are larger uncoated plates (20-60 nm). Acute toxicity (mortality at 48 or 96 h, depending on the test-organism) was assessed for the four species, from 0.1 to 100 mg L(-1) of NPs. Sub-lethal effects were assessed on chironomids exposed between 0.01 and 1 mg L(-1) of NPs. Mortality, growth inhibition and genotoxic effects were evaluated on amphibian larvae from 0.1 to 10 mg L(-1). Results reveal that no acute toxicity occurs on any species after short exposures, even at the highest concentrations. Mortality (35%) is observed on Xenopus larvae after 12d of exposure at the highest concentration of one type of NPs. No significant effects were observed on chironomids during chronic exposure. Xenopus larvae growth was inhibited from 1 mg L(-1) of both NPs while growth inhibition is observed on Pleurodeles only at the highest concentration of one type of NPs. No genotoxicity was observed on Xenopus but Pleurodeles exhibited dose-dependent genotoxic effects when exposed to one type of NPs. Observed differences in toxicity are discussed focusing on the studied compartment, routes of exposure, species and NPs., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The potential impact of Multiwalled Carbon NanoTubes (MWCNTs) was investigated on Xenopus laevis tadpoles exposed to 0.1, 1 and 10mg/L. Oxidative stress was measured in entire larvae exposed and DNA damage (Comet assay) was carried out in erythrocytes of circulating blood from 2h to 24h according to standardized recommendations. Results showed significant H2O2 production when larvae were exposed to 1mg/L and 10mg/L of MWCNTs after 4h and 2h of exposure, respectively. Antioxidant enzyme activities showed significant induction of catalase (CAT), glutathione reductase (GR) and superoxide dismutase (SOD) from only 2h of exposure to 10mg/L of MWCNTs. In presence of 1mg/L of MWCNTs, only GR and CAT activities were significantly induced at 4h. Enzyme activities do not follow a simple dose-effect relation, but the time of induction is shortened in relation with the tested concentration. The Comet assay results showed significant DNA damages with a dose dependent response. The profiles of DNA damages show fluctuations, in course of time, which are characteristics of oxidative stress response in relation with the continuous balance between damage and compensation process., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Insensitive acetylcholinesterase resistance due to a mutation in the acetylcholinesterase (ace) encoding ace-1 gene confers cross-resistance to organophosphate and carbamate insecticides in Anopheles gambiae populations from Central and West Africa. This mutation is associated with a strong genetic cost revealed through alterations of some life history traits but little is known about the physiological and behavioural changes in insects bearing the ace-1(R) allele. Comparative analysis of the salivary gland contents between An. gambiae susceptible and ace-1(R) resistant strains was carried out to charaterize factors that could be involved in modifications of blood meal process, trophic behaviour or pathogen interaction in the insecticide-resistant mosquitoes. Differential analysis of the salivary gland protein profiles revealed differences in abundance for several proteins, two of them showing major differences between the two strains. These two proteins identified as saglin and TRIO are salivary gland-1 related proteins, a family unique to anopheline mosquitoes, one of them playing a crucial role in salivary gland invasion by Plasmodium falciparum sporozoites. Differential expression of two other proteins previously identified in the Anopheles sialome was also observed. The differentially regulated proteins are involved in pathogen invasion, blood feeding process, and protection against oxidation, relevant steps in the outcome of malaria infection. Further functional studies and insect behaviour experiments would confirm the impact of the modification of the sialome composition on blood feeding and pathogen transmission abilities of the resistant mosquitoes. The data supports the hypothesis of alterations linked to insecticide resistance in the biology of the primary vector of human malaria in Africa.

Objective: Using human IgG antibody response to the Aedes Nterm-34 kDa salivary peptide as an indicator of human exposure to Aedes bites in surveying exposed populations from areas at risk of dengue virus (DENV) transmission in urban settings of Vientiane city, Lao PDR., Methods: Enzyme-linked immunosorbent assay tests were performed to measure the IgG response to Nterm-34 kDa peptide in blood samples collected within a flavivirus seroprevalence survey carried out in 2006 including 3558 randomly selected individuals. The level of IgG response to the Nterm-34 kDa peptide in individuals was analysed in relation to the level of urbanisation of the individual's residence, areas that presented significant differences in the prevalence of recent DENV infection., Results: No differences were observed in the anti-Nterm-34 kDa IgG level between DENV-positive and DENV-negative individuals. However, the level of specific IgG response was higher among individuals living in slightly urbanised neighbourhoods than among those in more highly urbanised areas (P < 0.0001). Interestingly, a similar pattern had already been observed concerning the prevalence of recent DENV infection in the same populations., Conclusion: The results of this retrospective study indicate that the evaluation of human IgG response to the Aedes Nterm-34 kDa salivary peptide could be a useful indicator to identify places with risk of dengue virus transmission in urban endemic areas., (© 2014 The Authors. Tropical Medicine & International Health Published by John Wiley & Sons Ltd.)

The diagnosis of childhood active tuberculosis (aTB) and latent Mycobacterium tuberculosis (M. tuberculosis) infection (LTBI) remains a challenge, and the replacement of tuberculin skin tests (TST) with commercialized gamma interferon (IFN-γ) release assays (IGRA) is not currently recommended. Two hundred sixty-six children between 1 month and 15 years of age, 214 of whom were at risk of recent M. tuberculosis infection and 51 who were included as controls, were prospectively enrolled in our study. According to the results of a clinical evaluation, TST, chest X ray, and microbiological assessment, each children was classified as noninfected, having LTBI, or having aTB. Long-incubation-time purified protein derivative (PPD), ESAT-6, and CFP-10 IGRA were performed and evaluated for their accuracy in correctly classifying the children. Whereas both TST and PPD IGRA were suboptimal for detecting aTB, combining the CFP-10 IGRA with a TST or with a PPD IGRA allowed us to detect all the children with aTB with a specificity of 96% for children who were positive for the CFP-10 IGRA. Moreover, the combination of the CFP-10 IGRA and PPD IGRA detected 96% of children who were eventually classified as having LTBI, but a strong IFN-γ response to CFP-10 (defined as >500 pg/ml) was highly suggestive of aTB, at least among the children who were <3 years old. The use of long-incubation-time CFP-10 IGRA and PPD IGRA should help clinicians to quickly identify aTB or LTBI in young children.

Aedes borne viruses represent public health problems in southern countries and threat to emerge in the developed world. Their control is currently based on vector population control. Much effort is being devoted to develop new tools to control such arbovirus. Recent findings suggest that the evaluation of human antibody (Ab) response to arthropod salivary proteins is relevant to measuring the level of human exposure to mosquito bites. Using an immunoepidemiological approach, the present study aimed to assess the usefulness of the salivary biomarker for measuring the efficacy of Ae. albopictus control strategies in La Reunion urban area. The antisaliva Ab response of adult humans exposed to Ae. albopictus was evaluated before and after vector control measures. Our results showed a significant correlation between antisaliva Ab response and the level of exposure to vectors bites. The decrease of Ae. albopictus density has been detected by this biomarker two weeks after the implementation of control measures, suggesting its potential usefulness for evaluating control strategies in a short time period. The identification of species specific salivary proteins/peptides should improve the use of this biomarker.

Aedes mosquitoes are important vectors of re-emerging diseases in developing countries, and increasing exposure to Aedes in the developed world is currently a source of concern. Given the limitations of current entomologic methods, there is a need for a new effective way for evaluating Aedes exposure. Our objective was to evaluate specific antibody responses to Aedes aegypti saliva as a biomarker for vector exposure in a dengue-endemic urban area. IgG responses to saliva were strong in young children and steadily waned with age. Specific IgG levels were significantly higher in persons living in sites with higher Ae. aegypti density, as measured by using entomologic parameters. Logistic regression showed a significant correlation between IgG to saliva and exposure level, independently of either age or sex. These results suggest that antibody responses to saliva could be used to monitor human exposure to Aedes bites.

Background: Much effort is being devoted for developing new indicators to evaluate the human exposure to Aedes mosquito bites and the risk of arbovirus transmission. Human antibody (Ab) responses to mosquito salivary components could represent a promising tool for evaluating the human-vector contact., Methodology/principal Findings: To develop a specific biomarker of human exposure to Aedes aegypti bites, we measured IgG Ab response to Ae. aegypti Nterm-34 kDa salivary peptide in exposed children in 7 villages of Southern Benin (West Africa). Results showed that specific IgG response presented high inter-individual heterogeneity between villages. IgG response was associated with rainfall and IgG level increased from dry (low exposure) to rainy (high exposure) seasons. These findings indicate that IgG Ab to Nterm-34 kDa salivary peptide may represent a reliable biomarker to detect variation in human exposure to Ae. aegypti bites., Conclusion/significance: This preliminary study highlights the potential use of Ab response to this salivary peptide for evaluating human exposure to Ae. aegypti. This biomarker could represent a new promising tool for assessing the risk of arbovirus transmission and for evaluating the efficacy of vector control interventions.

Human antibody (Ab) response to Anopheles whole saliva, used as biomarker of Anopheles exposure, was investigated over a period of two years (2008-2009), in children between 2 to 9 years old, before and after the introduction of three different malaria vector control methods; deltamethrin treated long lasting impregnated nets (LLIN) and insecticide treated plastic sheeting (ITPS)--Zero Fly®) (ITPS-ZF), deltamethrin impregnated Durable (Wall) Lining (ITPS-DL--Zerovector®) alone, and indoor residual spraying (IRS) with lambdacyhalothrin alone. These different vector control methods resulted in considerable decreases in all three entomological (82.4%), parasitological (54.8%) and immunological criteria analyzed. The highest reductions in the number of Anopheles collected and number of positive blood smears, respectively 82.1% and 58.3%, were found in Capango and Canjala where LLIN and ITPS-ZF were implemented. The immunological data based on the level of anti-saliva IgG Ab in children of all villages dropped significantly from 2008 to 2009, except in Chissequele. These results indicated that these three vector control methods significantly reduced malaria infections amongst the children studied and IRS significantly reduced the human-Anopheles contact. The number of Anopheles, positive blood smears, and the levels of anti-saliva IgG Ab were most reduced when LLIN and ITPS-ZF were used in combination, compared to the use of one vector control method alone, either ITPS-DL or IRS. Therefore, as a combination of two vector control methods is significantly more effective than one control method only, this control strategy should be further developed at a more global scale.

Background: The spread of Aedes albopictus, a vector for re-emergent arbovirus diseases like chikungunya and dengue, points up the need for better control strategies and new tools to evaluate transmission risk. Human antibody (Ab) responses to mosquito salivary proteins could represent a reliable biomarker for evaluating human-vector contact and the efficacy of control programs., Methodology/principal Findings: We used ELISA tests to evaluate specific immunoglobulin G (IgG) responses to salivary gland extracts (SGE) in adults exposed to Aedes albopictus in Reunion Island. The percentage of immune responders (88%) and levels of anti-SGE IgG Abs were high in exposed individuals. At an individual level, our results indicate heterogeneity of the exposure to Aedes albopictus bites. In addition, low-level immune cross-reactivity between Aedes albopictus and Aedes aegypti SGEs was observed, mainly in the highest responders., Conclusion/significance: Ab responses to saliva could be used as an immuno-epidemiological tool for evaluating exposure to Aedes albopictus bites. Combined with entomological and epidemiological methods, a "salivary" biomarker of exposure to Aedes albopictus could enhance surveillance of its spread and the risk of arbovirus transmission, and could be used as a direct tool for the evaluation of Aedes albopictus control strategies.

Populations of widely distributed species can be subjected to unequal selection pressures, producing differences in rates of local adaptation. We report a laboratory experiment testing tolerance variation to UV-B and polycyclic aromatic hydrocarbons (PAHs) among common frog (Rana temporaria) populations according to their natural exposure level in the field. Studied populations were naturally distributed along two gradients, i.e. UV-B radiation with altitude and level of contamination by PAHs with the distance to emitting sources (road traffic). Tadpoles from eight populations were subjected to (1) no or high level of artificial UV-B; (2) four concentrations of benzo[a]pyrene (BaP) (0, 50, 250, 500microgL(-1)); (3) simultaneously to UV-B and BaP. Since both stressors are genotoxic, the number of micronucleated erythrocytes (MNE) in circulating red blood cells was used as a bioindicator of tadpole sensitivity. High-altitude populations appear to be locally adapted to better resist UV-B genotoxicity, as they showed the lowest MNE numbers. Conversely, no correlation was observed between levels of PAH contamination in the field and tadpole tolerance to BaP in the laboratory, indicating the absence of local adaptation for BaP tolerance in these populations. Nevertheless, the decrease of MNE formation due to BaP exposure with altitude suggests that high-altitude populations were intrinsically more resistant to BaP genotoxicity. We propose the hypothesis of a co-tolerance between UV-B and BaP in high-altitude common frog populations: local adaptation to prevent and/or repair DNA damage induced by UV-B could also protect these highland populations against DNA damage induced by BaP. The results of this study highlight the role of local adaptation along pollutant gradients leading to tolerance variation, which implies that is it necessary to take into account the history of exposure of each population and the existence of co-tolerance that can hide toxic effects of a new pollutant.

Objective: To assess the extent of nosocomial transmission of tuberculosis among infants, family members, and healthcare workers (HCWs) who were exposed to a 29-week-old premature infant with congenital tuberculosis, diagnosed at 102 days of age., Design: A prospective exposure investigation using tuberculin skin test (IST conversion was conducted. Contacts underwent two skin tests 10 to 12 weeks apart. Clinical examination and chest radiographs were performed to rule out disease. Isoniazid prophylaxis was administered to exposed infants at higher risk., Setting: A neonatal intensive care unit in an urban hospital in Brussels, Belgium., Participants: Ninety-seven infants, 139 HCWs, and 180 visitors., Results: Newly positive TST results occurred in HCWs who had been in close contact with the infant. Six (19%) of 32 primary care nurses and physicians had TST conversions and received treatment. Among the 97 exposed infants, 85 were screened and 34 were identified as at higher risk of infection. Of these, 27 received preventive isoniazid. None of the infants and none of the 93 other infants' family members evaluated were infected., Conclusions: Congenital tuberculosis in an infant poses a risk for nosocomial transmission to HCWs. Delayed diagnosis of this rare disease and close proximity are the most important factors related to transmission.