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Two sequential batch reactors (R1 and R2) of aerobic granular sludge (AGS) were inoculated with activated sludge of different origins. The objective was to investigate the granulation and the consistency between the structure of the microbial communities (16S rRNA amplicon sequencing) in each reactor and their metabolic performance (removal of C, N, and P). Both reactors were fed with acetate-based synthetic wastewater, targeting an anaerobic-aerobic cycle reputed to favor the phosphorus- and glycogen-accumulating organisms (PAO and GAO). Stable granulation was achieved in both reactors, where, instead of PAO, the dominant genera were ordinary heterotrophic organisms (OHO) such as Thauera, Paracoccus, and Flavobacterium known for their high capacity of aerobic storage of polyhydroxyalkanoates (PHA). Generally, there was good consistency between the metabolic behavior of each reactor and the bacterial genera detected. Both reactors showed high removals of C and complete nitrification (Nitrosomonas and Nitrospira detected) but a low level of simultaneous nitrification-denitrification (SND) during the aerated phase. The latter causes that nitrates were recycled to the initial phase, in detriment of PAO selection. Meanwhile, the study showed that selecting slow-growing OHOs (with aerobic storage capacity) favors stable granulation, revealing an alternative AGS technology for C and N removal.

Dengue virus (DENV) was detected in Senegal in 1979 for the first time. Since 2017, unprecedented frequent outbreaks of DENV were noticed yearly. In this context, epidemiological and molecular evolution data are paramount to decipher the virus diffusion route. In the current study, we focused on a dengue outbreak which occurred in Senegal in 2018 in the context of a large religious gathering with 263 confirmed DENV cases out of 832 collected samples, including 25 life-threatening cases and 2 deaths. It was characterized by a co-circulation of dengue serotypes 1 and 3. Phylogenetic analysis based on the E gene revealed that the main detected serotype in Touba was DENV-3 and belonged to Genotype III. Bayesian phylogeographic analysis was performed and suggested one viral introduction around 2017.07 (95% HPD = 2016.61–2017.57) followed by cryptic circulation before the identification of the first case on 1 October 2018. DENV-3 strains are phylogenetically related, with strong phylogenetic links between strains retrieved from Burkina Faso and other West African countries. These phylogenetic data substantiate epidemiological data of the origin of DENV-3 and its spread between African countries and subsequent diffusion after religious mass events. The study also highlighted the usefulness of a mobile laboratory during the outbreak response, allowing rapid diagnosis and resulting in improved patient management.

Antimicrobial resistance (AMR) has become a global public health threat. Experts agree that unless proper actions are taken, the number of deaths due to AMR will increase. Many strategies are being pursued to tackle AMR, one of the most important being the development of efficient vaccines. Similar to other bacterial pathogens, AMR in

Dengue virus (DENV) was detected in Senegal in 1979 for the first time. Since 2017, unprecedented frequent occurrence of DENV outbreaks was noticed yearly. In this context, epidemiological and molecular evolution data are paramount to decipher virus diffusion route. In the current study, a dengue outbreak occurred in Senegal in 2018 in the context of the largest religious gathering with 263 confirmed DENV cases out of 832 collected samples, including 25 life-threatening cases and 2 deaths. It was characterized by a co-circulation of dengue serotypes 1 and 3. Phylogenetic analysis based on the E gene revealed that the main detected serotype in Touba city was DENV-3 and belonged to Genotype III. Bayesian phylogeographic analysis was performed and suggested one viral introduction around 2017.07 (95 % HPD = 2016.61 – 2017.57) followed by cryptic circulation before identification of the first case on 01 October 2018. DENV-3 strains are phylogenetically related, with strong phylogenetic links between strains retrieved from Burkina Faso and West African countries. These phylogenetic data substantiate epidemiological data of the DENV-3 origin and spread between African countries and subsequent diffusion after religious mass events. The study also highlighted the usefulness of the Mobile Laboratory during outbreak response allowing rapid diagnosis resulting in improved patient management.

Extra-intestinal pathogenic Escherichia coli (ExPEC), a predominant Gram-negative bacterial pathogen, express a wide range of virulence factors and is responsible of several diseases including urinary tract infections (UTI), nosocomial pneumonia, bacteremia, and neonatal meningitis. ExPEC isolates are often multidrug resistant (MDR) and clones producing extended-spectrum beta-lactamases (ESBL) are increasingly reported all over the world.Seventy-eight clinical ExPEC strains were selected for this study. The majority was from UTIs (n=51), while the rest (n=27) was from pus, sputum, bronchial fluid and vaginal samples (non-uropathogenic ExPEC). Interestingly, 49 out of the 78 ExPEC isolates where considered as community-acquired (CA) and 29 hospital-acquired (HA) bacteria. Antibiotic susceptibility testing was performed using the Kirby-Bauer disc diffusion method. Standard polymerase chain reaction (PCR) was used to screen major ESBL genes (blaCTX-M, blaOXA-1, blaTEM, blaSHV) and blaCTX-M variants (blaCTX-M-1, blaCTX-M-9, blaCTX-M-15, blaCTX-M-25).All the ExPEC strains were resistant to ampicillin, ticarcillin, amoxicillin/clavulanic acid combination, cefalotin, cefotaxime, ceftazidime, cefepime and aztreonam, but showed a high susceptibity to fosfomycin (98.7%, n = 77), ertapenem (96.2%, n = 75), and imipenem (100%). Moreover, isolates harbored at least one ESBL gene, including blaCTX-M (98.7%), blaOXA-1 (78.2%), blaTEM (44.9%) and blaSHV (3.8%). The CTX-M variants were also found with the predominance of blaCTX-M-1 (90.9%) and blaCTX-M-15 (90.9%) followed by blaCTX-M-9 (11.7%), while blaCTX-M-25 was not detected.Despite the resistance to most of the tested antibiotics, ExPEC isolates showed fortunately a good susceptibility to fosfomycin and carbapenems. blaCTX-M (blaCTX-M1, blaCTX-M15) and blaOXA-1 seem to be E. coli major ESBL genes circulating in Senegal. No significant difference was noted when comparing prevalence of ESBL genes detected from CA and HA strains, and from UPEC and non-uropathogenic ExPEC. The high level of resistance to antimicrobials observed stresses the need of establishing an epidemiological surveillance of antimicrobial resistance in both community and hospital settings.

Malaria is a serious public health burden in Senegal. Accurate diagnosis is essential to avoided unnecessary presumptive treatment. Malaria diagnosis currently relies on identifying malaria parasite using microscopy and detecting soluble parasite antigens by Rapid Diagnostic Tests (RDT). These techniques do not detect low-level, sub-patent malaria infections and are inherently hazardous and invasive. To overcome these obstacles, alternatives diagnostics method were explored. In contrast to blood, saliva presents a reduced biohazard and can be painlessly collected in relatively large quantities by individuals with moderate training. The objective of this study was to use saliva collected in OmnIGEN Kits as an alternative sample for malaria detection. Methods: A total of 77 febril patients tested malaria positive by mRDT were enrolled in this study. From each patient, blood sample was collected for dried blood spot and blood smear; and saliva sample on OMNIGEN kit. Parasite density was determined from smear and Plasmodium falciparum DNA was extracted from both dried blood spot and saliva samples collected from the same patients. Extracted DNA was amplied by qPCR machine. Results: Malaria prevalence using qPCR was 98,7% and 60% respectively with blood and saliva. Compared to blood, saliva showed a sensitivity; specicity; positive predictive value of 60%; 100 %; and 100 %. The concordance between parasites detection from saliva and blood was (p = 0.45). In addition, no difference was found between these two methods and the microscopy counting. Conclusions:Saliva could be a non-invasive alternative method for P. falciparum detection and epidemiological surveillance in country with limited ressources.

Molasses wastewaters (MWWs) such as baker’s yeast and distilleries effluents are usually pretreated by anaerobic, followed by aerobic biodegradation. This removes almost all the BOD, enough for meeting the current discharge standards in many low- and middle-income countries. However, as shown in the present work, the biotreated effluent still contains high levels of recalcitrant COD, color (melanoidins) and inorganic salts that end up in rivers (approx. 1000 mg/L COD, 2850 Pt–Co color units, 5000 mg/L TDS and 5400 μS/cm conductivity). To address this global problematics, and given the lack of proven cost-effective advanced treatment trains for MWW, this study assessed the performance and limits of ozonation and activated carbon (AC) adsorption (contrasting with O3 applications on raw MWW). The applied versus the reacted O3 doses were quantified, allowing also to estimate the ozone uptake rate as a new tool for characterizing the reactivity of the wastewaters. The effects of the treatments on different key parameters were studied: COD, color, aromaticity (UV254 nm), toxicity (Microtox) and biodegradability (by respirometry). O3 reduced the color (> 95%), but causing low COD mineralization (

In the present work, a laboratory scale anaerobic upflow reactor (UASB) was operated with a hydraulic retention time (6 h) and at psychrophilic temperature (< 20 oC). The UASB reactor treated wastewater from a chocolate industry with two applied organic volumetric loads (OLR apl ). Each OLR apl corresponded to an experimental stage: experimental stage I with 1.8 ± 0.8 kg soluble oxygen chemical demand ([DQO s ]/m 3 /d) and experimental stage II with 3.5 ± 0.9 kg COD s /m 3 /d. The objective of this research was to evaluate the removal efficiency of CODs (RE), biogas production (BP) and experimental cell yield ( Y exp ) within the laboratory scale UASB reactor. The control parameters monitored were pH, temperature and flow. Alkalinity, volatile fatty acids and COD s were analyzed in the influent and effluent, while suspended solids were analyzed inside the reactor and in the effluent. The RE during the steady state of the two experimental stages was greater than 90 %. The high RE obtained showed that the UASB reactor efficiently treated the industrial wastewater at low temperature with the OLR apl . BP increased when OLR apl increased (1.6 ± 0.8 and 3.5 ± 1.3 L/d of OLR apl of 1.8 and 3.5 kg COD s /m 3 /d, respectively [1 atm, 273 K]). The Y exp of experimental stage II was high (0.11 kg SSV/kg COD rem ). The high Y exp of experimental stage II could be due to the influence of psychrophilic temperature in the growth rate of anaerobic microorganisms.

Significance The threat to public health that is presented by zoonotic spillover of pathogens from animal reservoirs is predicted to increase with rapid anthropogenic changes and global trends such as migration and changing land use. Schistosomiasis currently infects more than 220 million people worldwide, and the multihost Schistosoma spp. system within Africa is a key example of where spillover of animal parasites into human populations has enabled the formation of viable hybrid parasite genotypes. Our study demonstrates how zoonotic spillover and complex interactions between pathogen species, such as parasite hybridization, may have implications such as resilience to current disease control strategies and may facilitate the spread of tropical diseases such as schistosomiasis beyond their original geographical boundaries., Zoonotic spillover and hybridization of parasites are major emerging public and veterinary health concerns at the interface of infectious disease biology, evolution, and control. Schistosomiasis is a neglected tropical disease of global importance caused by parasites of the Schistosoma genus, and the Schistosoma spp. system within Africa represents a key example of a system where spillover of animal parasites into human populations has enabled formation of hybrids. Combining model-based approaches and analyses of parasitological, molecular, and epidemiological data from northern Senegal, a region with a high prevalence of schistosome hybrids, we aimed to unravel the transmission dynamics of this complex multihost, multiparasite system. Using Bayesian methods and by estimating the basic reproduction number (R0), we evaluate the frequency of zoonotic spillover of Schistosoma bovis from livestock and the potential for onward transmission of hybrid S. bovis × S. haematobium offspring within human populations. We estimate R0 of hybrid schistosomes to be greater than the critical threshold of one (1.76; 95% CI 1.59 to 1.99), demonstrating the potential for hybridization to facilitate spread and establishment of schistosomiasis beyond its original geographical boundaries. We estimate R0 for S. bovis to be greater than one in cattle (1.43; 95% CI 1.24 to 1.85) but not in other ruminants, confirming cattle as the primary zoonotic reservoir. Through longitudinal simulations, we also show that where S. bovis and S. haematobium are coendemic (in livestock and humans respectively), the relative importance of zoonotic transmission is predicted to increase as the disease in humans nears elimination.

Background: In Senegal, malaria morbidity has shapely felt down over these past years. However, malaria epidemiology remains heterogeneous with persisting transmission in the southeastern part of the country and more important number of cases arising among older children and adolescents. Little is known about factors associated with clinical malaria among this group. A better understanding of malaria transmission among this new vulnerable group will guide future interventions targeting these key populations. This study aimed to identify factors associated with clinical malaria among adolescents in Senegal. Methods: A case control study was conducted from November to December 2020 in four health posts located in Saraya district. Cases were defined as adolescents (10-19 years) with uncomplicated malaria episode with fever (Temperature>37.5°) or history of fever and a positive malaria RDT. Controls were from the same age group, living in the neighborhood of the case, presenting a negative RDT. A standardized, pre-tested questionnaire was administered to each participant followed by home visit to assess participant’s living conditions. Factors associated with clinical malaria was assessed using a Stepwise Logistic regression analysis.Results: In total, 492 individuals were recruited (246 cases and 246 controls). In a multivariate analysis, factors associated with clinical malaria included non-use of bed net (aOR=2.65; 95% CI =1.58 - 4.45), non-use of other preventive measures (aOR=2.51; 95% CI=1.53 - 4.11) and indoor sleeping (aOR=3.22; 95%CI =1.66- 6.23). Protective factors included age of 15-19 years (aOR=0.38; 95% CI 0.23 - 0.62), absence of stagnant water around the house (aOR=0.27; 95% CI=0.16 - 0.44), having a female as head of household (aOR=0.47; 95% CI=0.25 - 0.90), occupation such as apprentice (OR=0.24; 95%CI=0.11 - 0.52).Conclusions: The study revealed that environmental factors and non-use of malaria preventive measures are the main determinant of malaria transmission among adolescents living in areas with persisting malaria transmission in Senegal. Strategies aiming at improving disease awareness and access to health care interventions such as LLIN are thus needed to improve malaria control and prevention among these vulnerable groups.

Hybridization of infectious agents is a major emerging public and veterinary health concern at the interface of evolution, epidemiology, and control. Whilst evidence of the extent of hybridization amongst parasites is increasing, their impact on morbidity remains largely unknown. This may be predicted to be particularly pertinent where parasites of animals with contrasting pathogenicity viably hybridize with human parasites. Recent research has revealed that viable zoonotic hybrids between human urogenital Schistosoma haematobium with intestinal Schistosoma species of livestock, notably Schistosoma bovis, can be highly prevalent across Africa and beyond. Examining human populations in Senegal, we found increased hepatic but decreased urogenital morbidity, and reduced improvement following treatment with praziquantel, in those infected with zoonotic hybrids compared to non-hybrids. Our results have implications for effective monitoring and evaluation of control programmes, and demonstrate for the first time the potential impact of parasite hybridizations on host morbidity.

Background Dengue fever is a mosquito born disease associated with self-limited to life threatening illness. First detected in Senegal in the nineteenth century, and despite its growing incidence this last decade, significant knowledge gaps exist in our knowledge of genetic diversity of circulating strains. This study highlights the circulating serotypes and genotypes between January 2017 and December 2018 and their spatial and temporal distribution throughout all regions of Senegal. Methods We used 56 dengue virus (DENV) strains for the analysis collected from 11 sampling areas: 39 from all regions of Senegal, and 17 isolates from Thiès, a particular area of the country. Two real time RT-qPCR systems were used to confirm dengue infection and corresponding serotypes. For molecular characterization, CprM gene was sequenced and submitted to phylogenetic analysis for serotypes and genotypes assignment. Results Three dengue virus serotypes (DENV-1–3) were detected by all used methods. DENV-3 was detected in 50% (28/56) of the isolates, followed by DENV-1 and DENV-2, each representing 25% (14/56) of the isolates. DENV-3 belongs to genotype III, DENV-1 to genotype V and DENV-2 to Cosmopolitan genotype. Serotype 3 was detected in 7 sampling locations and a co-circulation of different serotypes was observed in Thiès, Fatick and Richard-toll. Conclusions These results emphasize the need of continuous DENV surveillance in Senegal to detect DENV cases, to define circulating serotypes/genotypes and to prevent the spread and the occurrence of severe cases.

The structural effect of Fe-, Zr- and FeZr-modified clinoptilolite-, mordenite- and chabazite-rich tuffs was examined on the As(V) sorption from aqueous solutions. Batch experiments were conducted considering the sorbent mass, contact time and pH. The most probable sites where Fe and Zr species could be located in the zeolite frameworks after the modification were described: Na+ exchangeable sites in the 10-ring and 8-ring channels of clinoptilolite, and inside the 12-ring and 8-ring channels and 8-ring side pockets of mordenite; Na+, K+ and Ca2+ sites within the D6R cages and barrel-shape cavities of chabazite. As(V) sorption capacities of Zr-modified materials (0.062 ± 0.008 mg/g) were higher than the Fe-modified ones (0.031 ± 0.007 mg/g). Isotherm results were best adjusted to the Freundlich model; FeZr-modified materials presented the highest affinity for arsenic. Estimated free energy values from the Dubinin-Radushkevich isotherm model suggested an ion exchange process. Kinetic data were well described by the pseudo-second order and intraparticle diffusion models, thus the sorption rate could have been controlled by two steps, the transfer of arsenic through the channel systems and the chemical interactions. Based on data modeling for the pH effect, zeolitic materials showed an optimal sorption in the pH range from 2 to 6; the major arsenic species involved in the sorption process were H2AsO4− and HAsO42−. The mechanism for As(V) removal by the FeZr-modified zeolites was through the formation of an inner sphere complex via ion exchange.

Summary Background Schistosomiasis is a neglected tropical disease of global medical and veterinary importance. As efforts to eliminate schistosomiasis as a public health problem and interrupt transmission gather momentum, the potential zoonotic risk posed by livestock Schistosoma species via viable hybridisation in sub-Saharan Africa have been largely overlooked. We aimed to investigate the prevalence, distribution, and multi-host, multiparasite transmission cycle of Haematobium group schistosomiasis in Senegal, West Africa. Methods In this epidemiological study, we carried out systematic surveys in definitive hosts (humans, cattle, sheep, and goats) and snail intermediate hosts, in 2016–18, in two areas of Northern Senegal: Richard Toll and Lac de Guiers, where transmission is perennial; and Barkedji and Linguere, where transmission is seasonal. The occurrence and distribution of Schistosoma species and hybrids were assessed by molecular analyses of parasitological specimens obtained from the different hosts. Children in the study villages aged 5–17 years and enrolled in school were selected from school registers. Adults (aged 18–78 years) were self-selecting volunteers. Livestock from the study villages in both areas were also randomly sampled, as were post-mortem samples from local abattoirs. Additionally, five malacological surveys of snail intermediate hosts were carried out at each site in open water sources used by the communities and their animals. Findings In May to August, 2016, we surveyed 375 children and 20 adults from Richard Toll and Lac de Guiers, and 201 children and 107 adults from Barkedji and Linguere; in October, 2017, to January, 2018, we surveyed 386 children and 88 adults from Richard Toll and Lac de Guiers, and 323 children and 85 adults from Barkedji and Linguere. In Richard Toll and Lac de Guiers the prevalence of urogenital schistosomiasis in children was estimated to be 87% (95% CI 80–95) in 2016 and 88% (82–95) in 2017–18. An estimated 63% (in 2016) and 72% (in 2017–18) of infected children were shedding Schistosoma haematobium–Schistosoma bovis hybrids. In adults in Richard Toll and Lac de Guiers, the prevalence of urogenital schistosomiasis was estimated to be 79% (52–97) in 2016 and 41% (30–54) in 2017–18, with 88% of infected samples containing S haematobium–S bovis hybrids. In Barkedji and Linguere the prevalence of urogenital schistosomiasis in children was estimated to be 30% (23–38) in 2016 and 42% (35–49) in 2017–18, with the proportion of infected children found to be shedding S haematobium–S bovis hybrid miracidia much lower than in Richard Toll and Lac de Guiers (11% in 2016 and 9% in 2017–18). In adults in Barkedji and Linguere, the prevalence of urogenital schistosomiasis was estimated to be 26% (17–36) in 2016 and 47% (34–60) in 2017–18, with 10% of infected samples containing S haematobium–S bovis hybrids. The prevalence of S bovis in the sympatric cattle population of Richard Toll and the Lac de Guiers was 92% (80–99), with S bovis also found in sheep (estimated prevalence 14% [5–31]) and goats (15% [5–33]). In Barkedji and Linguere the main schistosome species in livestock was Schistosoma curassoni, with an estimated prevalence of 73% (48–93) in sheep, 84% (61–98) in goats and 8% (2–24) in cattle. S haematobium–S bovis hybrids were not found in livestock. In Richard Toll and Lac de Guiers 35% of infected Bulinus spp snail intermediate hosts were found to be shedding S haematobium–S bovis hybrids (68% shedding S haematobium; 17% shedding S bovis); however, no snails were found to be shedding S haematobium hybrids in Barkedji and Linguere (29% shedding S haematobium; 71% shedding S curassoni). Interpretation Our findings suggest that hybrids originate in humans via zoonotic spillover from livestock populations, where schistosomiasis is co-endemic. Introgressive hybridisation, evolving host ranges, and wider ecosystem contexts could affect the transmission dynamics of schistosomiasis and other pathogens, demonstrating the need to consider control measures within a One Health framework. Funding Zoonoses and Emerging Livestock Systems programme (UK Biotechnology and Biological Sciences Research Council, UK Department for International Development, UK Economic and Social Research Council, UK Medical Research Council, UK Natural Environment Research Council, and UK Defence Science and Technology Laboratory).

Background: Past and recent outbreaks have highlighted the vulnerability of humans to infectious diseases, which represent serious economic and health security threats. A paradigm shift in the management of sanitary crises is urgently needed. Based on lessons from the 2014 Ebola outbreak, the Praesens Foundation has developed an all-terrain mobile biosafety laboratory (MBS-Lab) for effective field diagnostics capabilities. Objective: The aim of the study was to train African teams and run a field evaluation of the MBS-Lab, including robustness, technical and operational sustainability, biosafety, connectivity, turn-around times for testing and result delivery. Methods: The MBS-Lab was deployed in Senegal in October 2017 for a six-month field assessment under various ecological conditions and was mobilised during the dengue outbreaks in 2017 and 2018. Results: The MBS-Lab can be considered an off-grid solution that addresses field challenges with regard to working conditions, mobility, deployment, environment and personnel safety. Blood (n = 398) and nasal swab (n = 113) samples were collected from 460 study participants for molecular screening for acute febrile illnesses and respiratory infections. The results showed that malaria (particularly in Kédougou) and upper respiratory tract infections remain problematic. Suspected dengue samples were tested on board during the dengue outbreaks in 2017 (882 tests; 128 confirmed cases) and 2018 (1736 tests; 202 confirmed cases). Conclusion: The MBS-Lab is an innovative solution for outbreak response, even in remote areas. The study demonstrated successful local ownership and community engagement. The MBS-Lab can also be considered an open mobile healthcare platform that offers various opportunities for field-deployable, point-of-care technologies for surveillance programmes.

In West Africa, Schistosoma spp. are capable of infecting multiple definitive hosts, a lifecycle feature that may complicate schistosomiasis control. We characterized the evolutionary relationships among multiple Schistosoma mansoni isolates collected from snails (intermediate hosts), humans (definitive hosts), and rodents (definitive hosts) in Senegal. On a local scale, diagnosis of S. mansoni infection ranged 3.8%-44.8% in school-aged children, 1.7%-52.6% in Mastomys huberti mice, and 1.8%-7.1% in Biomphalaria pfeifferi snails. Our phylogenetic framework confirmed the presence of multiple S. mansoni lineages that could infect both humans and rodents; divergence times of these lineages varied (0.13-0.02 million years ago). We propose that extensive movement of persons across West Africa might have contributed to the establishment of these various multihost S. mansoni clades. High S. mansoni prevalence in rodents at transmission sites frequented by humans further highlights the implications that alternative hosts could have on future public health interventions.

With the growing success of controlling malaria in Sub-Saharan Africa, the incidence of fever due to malaria is in decline, whereas the proportion of patients with non-malaria febrile illness (NMFI) is increasing. Clinical diagnosis of NMFI is hampered by unspecific symptoms, but early diagnosis is a key factor for both better patient care and disease control. The aim of this study was to determine the arboviral aetiologies of NMFI in low resource settings, using a mobile laboratory based on recombinase polymerase amplification (RPA) assays. The panel of tests for this study was expanded to five arboviruses: dengue virus (DENV), zika virus (ZIKV), yellow fever virus (YFV), chikungunya virus (CHIKV), and rift valley fever virus (RVFV). One hundred and four children aged between one month and 115 months were enrolled and screened. Three of the 104 blood samples of children

Molecular detection of Zika virus (ZIKV) is a key element of outbreak management. Multiple PCR and isothermal ZIKV assays targeting different ZIKV sequences have been published. In this study, we compared a qRT-PCR, 2 RT-LAMP assays (based on different primer design approaches), and an RT-RPA for the detection of African and Asian/American lineages of ZIKV isolates from human, mosquito, and monkey. Results showed that RT-LAMP detected 100% of samples with a time threshold (Tt) of 18.01 ± 11.71 min while qRT-PCR detected 88.88% of samples with a Tt of 58.30 ± 16.58 min and RT-RPA 50% of samples with a Tt of 3.70 ± 0.44 min.

Two one-step real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of Zika virus (ZIKV) were developed, based on two different primer design approaches: (1) open source, based on a combination of sequence diversity clustering (phylogeny and principal component analysis) and LAVA algorithm, using 45 whole genome ZIKV sequences retrieved from the National Center for Biotechnology Information (NCBI) database; (2) standard software for LAMP primer design (Primer Explorer V4), using 59 sequences of the ZIKV 3' UTR. The assays were firstly evaluated with External Quality Assessment panels from INSTAND e.V. (Germany) and EVD-LabNet (The Netherlands) including 4 and 12 unknown samples, respectively, and secondly, with 9 human, mosquito, and monkey ZIKV isolates from Africa (Senegal, Ivory Coast, and Uganda) and America (Brazil). The limit of detection as determined by probit analysis was 181 molecules for both RT-LAMP assays, and 100% reproducibility in the assays was obtained for 10

Background In this work, a platform was developed and tested to allow to detect a variety of candidate viral, bacterial and parasitic pathogens, for acute fever of unknown origin. The platform is based on a centrifugal microfluidic cartridge, the LabDisk (“FeverDisk” for the specific application), which integrates all necessary reagents for sample-to-answer analysis and is processed by a compact, point-of-care compatible device. Methodology/Principal findings A sample volume of 200 μL per FeverDisk was used. In situ extraction with pre-stored reagents was achieved by bind-wash-elute chemistry and magnetic particles. Enzymes for the loop-mediated isothermal amplification (LAMP) were pre-stored in lyopellet form providing stability and independence from the cold chain. The total time to result from sample inlet to read out was 2 h. The proof-of-principle was demonstrated in three small-scale feasibility studies: in Dakar, Senegal and Khartoum, Sudan we tested biobanked samples using 29 and 9 disks, respectively; in Reinfeld, Germany we tested spiked samples and analyzed the limit of detection using three bacteria simultaneously spiked in whole blood using 15 disks. Overall during the three studies, the FeverDisk detected dengue virus (different serotypes), chikungunya virus, Plasmodium falciparum, Salmonella enterica Typhi, Salmonella enterica Paratyphi A and Streptococcus pneumoniae. Conclusions/Significance The FeverDisk proved to be universally applicable as it successfully detected all different types of pathogens as single or co-infections, while it also managed to define the serotype of un-serotyped dengue samples. Thirty-eight FeverDisks at the two African sites provided 59 assay results, out of which 51 (86.4%) were confirmed with reference assay results. The results provide a promising outlook for future implementation of the platform in larger prospective clinical studies for defining its clinical sensitivity and specificity. The technology aims to provide multi-target diagnosis of the origins of fever, which will help fight lethal diseases and the incessant rise of antimicrobial resistance., Author summary Infectious diseases in tropical regions may have a variety of viral, bacterial or parasitic origins and a patient may suffer from several diseases simultaneously, each presenting with acute fever as a clinical symptom. This makes it difficult to determine the origin of the pathogen causing the disease(s). In addition to the endemic infectious diseases, outbreaks of epidemics frequently complicate diagnostic demands. Accurate diagnosis for proper patient management requires the utilization of highly sensitive and specific, rapid, easy-to-use diagnostic tools compatible with point-of-care settings. We describe the use of a disk-shaped microfluidic platform, the “FeverDisk”, for differential diagnosis of acute fever. Our FeverDisk platform demonstrated its capability to detect bacteria, viruses and parasites that are typical of tropical single and co-infections from biobanked samples within only 2 hours and in very good agreement with reference method results. This, in combination with its easy-to-use and point-of-care compatible nature, render our platform a promising candidate for detection of tropical diseases and precise identification of the cause of acute fever, in endemic and epidemic settings. Future work will involve extensive clinical characterization of the platform in prospective studies.

Two one-step real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of Zika virus (ZIKV) were developed, based on two different primer design approaches: (1) open source, based on a combination of sequence diversity clustering (phylogeny and principal component analysis) and LAVA algorithm, using 45 whole genome ZIKV sequences retrieved from the National Center for Biotechnology Information (NCBI) database; (2) standard software for LAMP primer design (Primer Explorer V4), using 59 sequences of the ZIKV 3' UTR. The assays were firstly evaluated with External Quality Assessment panels from INSTAND e.V. (Germany) and EVD-LabNet (The Netherlands) including 4 and 12 unknown samples, respectively, and secondly, with 9 human, mosquito, and monkey ZIKV isolates from Africa (Senegal, Ivory Coast, and Uganda) and America (Brazil). The limit of detection as determined by probit analysis was 181 molecules for both RT-LAMP assays, and 100% reproducibility in the assays was obtained for 103 molecules (4/8 repetitions were positive for 102 molecules). Both assays were specific, amplifying only ZIKV RNA and not cross-detecting other arboviruses included in this study.

Molecular detection of Zika virus (ZIKV) is a key element of outbreak management. Multiple PCR and isothermal ZIKV assays targeting different ZIKV sequences have been published. In this study, we compared a qRT-PCR, 2 RT-LAMP assays (based on different primer design approaches), and an RT-RPA for the detection of African and Asian/American lineages of ZIKV isolates from human, mosquito, and monkey. Results showed that RT-LAMP detected 100% of samples with a time threshold (Tt) of 18.01 ± 11.71 min while qRT-PCR detected 88.88% of samples with a Tt of 58.30 ± 16.58 min and RT-RPA 50% of samples with a Tt of 3.70 ± 0.44 min.

El objetivo de este estudio fue determinar la toxicidad de los sedimentos del curso alto del rio Lerma (CARL), Estado de Mexico, por medio del ensayo Microtox® con el protocolo basico en la fase solida. Se colectaron muestras de sedimento superficial en 23 sitios distribuidos sobre el CARL: 12 sobre el cauce del rio y 11 en sus tributarios, durante dos campanas de muestreo en un periodo de un ano. La concentracion efectiva media (EC50) en Vibrio fischeri vario para los sedimentos riverinos entre 139 mg/L y 3588 mg/L y para los de las descargas fue entre 108 mg/L y 16436 mg/L. En general los sedimentos del CARL pueden clasificarse como extremadamente toxicos, lo cual es un indicativo del impacto de las contribuciones antropicas de origen domestico e industrial que recibe este importante cuerpo de agua.

The complex multi-host disease dynamics of schistosomiasis and Schistosoma spp., including the emergence of zoonotic parasite hybrids, remain largely unexplored in West Africa. We elucidated the role of wild small mammals as reservoir for zoonotic Schistosoma species and hybrids in endemic areas of Senegal. We identified Schistosoma mansoni, Schistosoma bovis, and a Schistosoma haematobium/S. bovis hybrid, with local prevalence in wild rodents ranging from 1.9% to 28.6%. Our findings indicate that rodents may be an important local reservoir for zoonotic schistosomiasis in endemic areas of West Africa, amplifying transmission to humans and acting as natural definitive hosts of schistosome hybrids.

Applying ozone to the return flow in an activated sludge (AS) process is a way for reducing the residual solids production. To be able to extend the activated sludge models to the ozone-AS process, adequate prediction of the tri-atoms effects on the particulate COD fractions is needed. In this study, the biomass inactivation, COD mineralization, and solids dissolution were quantified in batch tests and dose-response models were developed as a function of the reacted ozone doses (ROD). Three kinds of model-sludge were used. S1 was a lab-cultivated synthetic sludge with two components (heterotrophs XH and XP). S2 was a digestate of S1 almost made by the endogenous residues, XP. S3 was from a municipal activated sludge plant. The specific ozone uptake rate (SO3UR, mgO3/gCOD.h) was determined as a tool for characterizing the reactivity of the sludges. SO3UR increased with the XH fraction and decreased with more XP. Biomass inactivation was exponential (e-β.ROD) as a function of the ROD doses. The percentage of solids reduction was predictable through a linear model (CMiner + Ysol ROD), with a fixed part due to mineralization (CMiner) and a variable part from the solubilization process. The parameters of the models, i.e. the inactivation and the dissolution yields (β, 0.008-0.029 (mgO3/mgCODini)-1 vs Ysol, 0.5-2.8 mg CODsol/mgO3) varied in magnitude, depending on the intensity of the scavenging reactions and potentially the compactness of the flocs for each sludge.