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Over the last two decades, an increase in bed bug infestations has been observed worldwide. Although their definitive role as vectors of infectious agents has not yet been demonstrated, bed bugs have a direct effect on human health through dermatological reactions to their bites and psychological disorders linked to domestic infestations. In this study, the effectiveness of using MALDI-TOF MS to correctly identify these two bed bug species at immature stages was assessed, as well as it effectiveness as discriminating between the immature stages (IS) of C. lectularius and C. hemipterus and their associated developmental stages. A total of 305 specimens were subjected to MALDI-TOF MS analysis, including 153 C. lectularius (28 eggs and 25 nymphs per stage from IS1 to IS5) and 152 C. hemipterus (27 eggs and 25 nymphs per stage from IS1 to IS5). ). MALDI-TOF MS analysis enabled us to obtain 84.97% (130/153) of high-quality MS spectra in terms of reproducibility and profile intensity. Twenty-four spectra including two per stage, from egg to IS5, and per bed bug species - were added to our in-house MS reference arthropod spectra database. All specimens were correctly identified at the species level, independently of the developmental stage, with log score values (LSVs) ranging from 1.75 to 2.79 (mean = 2.29 ± 0.12) and 1.81 to 2.71 (mean = 2.37 ± 0.03) for C. lectularius and C. hemipterus, respectively. MALDI-TOF MS correctly classified 53,33% (104/195) of the Cimex at the correct immature stage. Conversely, an accurate comparison of the profiles with a Genetic Algorithm model underlined that grouping the immature stages in two groups, early (IS1-IS2) and late (IS3-IS4-IS5), made it possible to obtain a cross validation (CV) and recognition capability (RC) greater than 92% and 94%, respectively, for both species. This study holds great promise for the management of bed bug infestations., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Termites are one of the most common pests that damage wood and other cellulosic materials . Although Africa has more varieties of termite species than any other continent, few entomological studies have been conducted in Gabon. Identifying termites poses significant difficulties for entomologists. The aim of this study was to evaluate the reliability and confirm the significance of MALDI-TOF MS in identifying fresh termites collected in equatorial Africa. A total of 108 termites were collected from 13 termite nests during a field mission in 2021 in Lekedi and Bongoville, Gabon. Termites were morphologically identified and subjected to MALDI-TOF MS, then molecular analyses using the COI and 12S rRNA genes. Four termite species were morphologically identified in this study: Pseudacanthotermes militaris , Macrotermes muelleri , Macrotermes nobilis , and Noditermes indoensis . However, when using molecular biology, only three species were identified, namely Macrotermes bellicosus , P. militaris , and N. indoensis, because the specimens initially identified as M. muelleri and M. nobilis were found to be M. bellicosus . The MALDI-TOF MS spectral profiles of the termites were all of good quality, with intra-species reproducibility and inter-species specificity. The spectra of 98 termites were blind tested against our upgraded database, which included the spectra of ten termite specimens. All tested spectra were correctly matched to their respective species, with log score values (LSVs) ranging from 1.649 to 2.592. The mean LSV was 2.215 ± 0.203, and the median was 2.241. However, 95.91% (94/98) of our spectra had LSVs above 1.8. This study demonstrates how a proteomic approach can overcome termites' molecular and morphological identification limitations and serve as a useful taxonomic tool., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Hasnaoui reports financial support was provided by Mediterranean University Hospital Institute for Infectious Diseases., (© 2024 The Authors. Published by Elsevier Ltd.)

Bed bugs are considered a major public health problem in industrialized countries. Usually, bed bug infestations are managed using a combination of physical and chemical methods. In recent years, new strategies for bed bug control have emerged, particularly the use of dusts like diatomaceous earth and silicon dioxide. However, in Europe, the use of silicon dioxide is restricted to professional, while diatomaceous earth can be harmful to the lungs. This study aimed to assess bed bug mortality rates associated with Sommières earth, green clay, talc, and sodium bicarbonate compared to silicon dioxide and diatomaceous earth from a pest management company, diatomaceous earth for litter conditioner, and diatomaceous earth from a supermarket. We tested permanent exposure, short exposure, horizontal transfer and repellent effect on two bed bug colonies. Sommières earth demonstrated efficacy ranging from 75% to 100% in permanent and short exposures, similar to the efficacy of diatomaceous earth from the pest management company. On the contrary, diatomaceous earth for litter conditioner and diatomaceous earth from a supermarket, green clay, talc, and sodium bicarbonate were found to be ineffective. This study demonstrates, for the first time, the efficacy of Sommières earth against bed bugs, but also highlights the variability in efficacy of diatomaceous earths on bed bugs depending on their quality., (© H. Almou Oumarou et al., published by EDP Sciences, 2024.)

After vanishing from the public eye for more than 50 years, bed bugs have resurged to become one of the most widely discussed and heavily researched insect pests in the world. This study presents the basic information of infestations of tropical bed bugs, Cimex hemipterus (Hemiptera: Cimicidae), in Cameroon. A total of 248 immature stage and adult bed bug specimens were collected from households and a travel agency in Yaoundé and Douala, Cameroon. The ability of MALDI-TOF MS to identify bed bugs was tested using heads for adults and cephalothoraxes for immature stages. Microorganism screening was performed by qPCR and confirmed by regular PCR and sequencing. Based on morphometrical criteria, four stages of immature bed bugs are represented. Of the 248 bed bug specimens morphologically identified as Cimex hemipterus, 246 (77 males, 65 females and 104 immature specimens) were submitted to MALDI-TOF MS analysis. Of the 222 adults and immature specimens tested, 122 (59.9 %) produced good quality MALDI-TOF MS spectra (35 adults and 87 immature specimens). Blind testing allowed species level identification of 98.21 % of adult and immature C. hemipterus. Among the bacteria tested, only Wolbachia DNA was found in 12/246 (4.8 %) bed bugs. More surveys in the country are warranted to assess the true level of bed bug infestations, in order to take appropriate action for their control., 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. Published by Elsevier B.V.)

Background: Many studies have evaluated the use of hydroxychloroquine in COVID-19. Most retrospective observational studies demonstrate a benefit of using HCQ on mortality, but not most randomized clinical trials., Methods: We analyzed raw data collected from a cohort of 30,423 patients with COVID-19 cared for at IHU Méditerranée Infection in Marseille France and extracted from the DRYAD open data platform. We performed univariate and multivariable logistic regressions with all-cause mortality within six weeks. Multivariable logistic regressions were adjusted for sex, age group (<50, 50-69, 70-89 and ​> ​89 years), periods (or variants), and type of patient management., Results: Among 30,202 patients for whom information on treatment was available, 191/23,172 (0.82%) patients treated with HCQ-AZ died, compared to 344/7030 (4.89%) who did not receive treatment with HCQ-AZ. HCQ-AZ therapy was associated with a lower mortality than treatment without HCQ-AZ (odds ratio (OR) 0.16; 95% confidence interval (CI), 0.14-0.19). After adjustment for sex, age, period, and patient management, HCQ-AZ was associated with a significantly lower mortality rate (adjusted OR (aOR) 0.55, 95% CI 0.45-0.68). On a subsample of 21,664 patients with available variant information, results remained robust after adjustment on sex, age, patient management and variant (aOR 0.55; 95% CI 0.44-0.69). On a subsample of 16,063 patients, HCQ-AZ was still associated with a significantly lower mortality rate (aOR 0.47, 95%CI 0.29-0.75) after adjustment for sex, age, period, patient management, vaccination status and comorbidities., Conclusion: Analysis of this large online database showed that HCQ-AZ was consistently associated with the lowest mortality., Competing Interests: The authors have completed the Unified Competing Interest form (available on request from the corresponding author). DR declares grants, contracts, royalties and/or licenses from Hitachi High-Technologies Corporation, Tokyo, Japan. DR is a scientific board member of Eurofins. DR is founder and shareholder of four startups, none which have yet generated an income: a microbial culture company (Culture Top), two biotechnology companies (Techno-Jouvence and Gene and Green TK), and a rapid diagnosis of infectious diseases company (Pocramé). PB, MM and PMC declare no support from any organization for the submitted work, no financial relationships with any organisations that might have an interest in the submitted work in the previous three years, and no other relationships or activities that could appear to have influenced the submitted work., (© 2023 The Authors. Published by Elsevier Ltd.)

Ticks are a significant group of arthropod vectors that transmit a large variety of pathogens responsible for human and animal diseases worldwide. Ticks are the second biggest transmitters of vector-borne diseases, behind mosquitoes. However, in West Africa, there is often only limited knowledge of tick-borne diseases. With the scarcity of appropriate diagnostic services, the prevalence of tick-borne diseases is generally underestimated in humans. In this review, we provide an update on tick-borne pathogens reported in people, animals and ticks in West Africa by microscopic, immunological and molecular methods. A systematic search was conducted in PubMed and Google Scholar. The selection criteria included all studies conducted in West Africa reporting the presence of Rickettsia , Borrelia , Anaplasma , Ehrlichia , Bartonella , Coxiella burnetii , Theileria , Babesia , Hepatozoon and Crimean-Congo haemorrhagic fever viruses in humans, animals or ticks. Our intention is to raise awareness of tick-borne diseases amongst human and animal health workers in West Africa, and also physicians working with tourists who have travelled to the region.

MALDI-TOF is now considered a relevant tool for the identification of arthropods, including lice and fleas. However, the duration and conditions of storage, such as in ethanol, which is frequently used to preserve these ectoparasites, could impede their classification. The purpose of the present study was to assess the stability of MS profiles from Pediculus humanus corporis lice and Ctenocephalides felis fleas preserved in alcohol from one to four years and kinetically submitted to MALDI-TOF MS. A total of 469 cephalothoraxes from lice (n = 170) and fleas (n = 299) were tested. The reproducibility of the MS profiles was estimated based on the log score values (LSVs) obtained for query profiles compared to the reference profiles included in the MS database. Only MS spectra from P. humanus corporis and C. felis stored in alcohol for less than one year were included in the reference MS database. Approximately 75% of MS spectra from lice (75.2%, 94/125) and fleas (74.4%, 122/164) specimens stored in alcohol for 12 to 48 months, queried against the reference MS database, obtained relevant identification. An accurate analysis revealed a significant decrease in the proportion of identification for both species stored for more than 22 months in alcohol. It was hypothesized that incomplete drying was responsible for MS spectra variations. Then, 45 lice and 60 fleas were subjected to longer drying periods from 12 to 24 h. The increase in the drying period improved the proportion of relevant identification for lice (95%) and fleas (80%). This study highlighted that a correct rate of identification by MS could be obtained for lice and fleas preserved in alcohol for up to four years on the condition that the drying period was sufficiently long for accurate identification.

During the current global outbreak of mpox (formerly monkeypox), atypical features were frequently described outside endemic areas, raising concerns around differential diagnosis. In this study, we included 372 adult patients who had clinical signs consistent with mpox and who were screened using non-variola orthopoxvirus specific quantitative polymerase chain reaction (PCR) between 15 May and 15 November 2022 at the University Hospital Institute Méditerranée Infection, Marseille, France. At least one clinical sample was positive for 143 (38.4%) of these patients and 229 (61.6%) were negative. Clinically, patients who had mpox presented more frequently with systemic signs (69.9% vs. 31.0%, p < 10 -6 ) including fever (51.0% vs. 30.1%, p < 10 -3 ), myalgia (33.5% vs. 17.9%, p = 0.002), and lymphadenopathy (38.5% vs. 13.1%, p < 10 -6 ). Among the patients who were negative for the non-variola orthopoxvirus, an alternative diagnosis was identified in 58 of them (25.3%), including chickenpox (n = 30, 13.1%), syphilis (n = 9, 4%), bacterial skin infection (n = 8, 3.5%), gonococcus (n = 5, 2.2%), HSV infection (n = 5, 2.2%), and histoplasmosis (n = 1, 0.4%). Overall, in the current outbreak, we show that mpox has a poorly specific clinical presentation. This reinforces the importance of microbiological confirmation. In symptomatic patients who are negative for the monkeypox virus by PCR, a broad differential diagnosis should be maintained., (© 2023 Wiley Periodicals LLC.)

Mosquitoes are arthropods that represent a real public health problem in Africa. Morphology and molecular biology techniques are usually used to identify different mosquito species. In recent years, an innovative tool, matrix-assisted desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), has been used to identify many arthropods quickly and at low cost, where equipment is available. We evaluated the ability of MALDI-TOF MS to identify mosquitoes collected in Senegal and stored for several months in silica gel, and to determine the origin of their blood meal. A total of 582 mosquitoes were collected and analysed. We obtained 329/582 (56.52%) MALDI-TOF MS good-quality spectra from mosquito legs and 123/157 (78.34%) good-quality spectra from engorged abdomens. We updated our home-made MALDI-TOF MS arthropod spectra database by adding 23 spectra of five mosquito species from Senegal that had been identified morphologically and molecularly. These included legs from Anopheles gambiae , Anopheles arabiensis, Anopheles cf. rivulorum , Culex nebulosus , Anopheles funestus , and three spectra from abdomens engorged with human blood. Having updated the database, all mosquitoes tested by MALDI-TOF MS were identified with scores greater than or equal to 1.7 as An. gambiae ( n = 64), Anopheles coluzzii ( n = 12), An. arabiensis ( n = 1), An. funestus ( n = 7), An. cf rivulorum ( n = 1), Lutzia tigripes ( n = 3), Cx. nebulosus ( n = 211), Culex quinquefasciatus ( n = 2), Culex duttoni ( n = 1), Culex perfescus ( n = 1), Culex tritaeniorhynchus ( n = 1), and Aedes aegypti ( n = 2). Blood meal identification by MALDI-TOF MS revealed that mosquitoes had fed on the blood of humans ( n = 97), cows ( n = 6), dogs ( n = 2), goats ( n = 1), sheep ( n = 1), and bats ( n = 1). Mixed meals were also detected. These results confirm that MALDI-TOF MS is a promising technique for identifying mosquitoes and the origin of their blood meal.

The soft ticks, Ornithodoros sonrai , are known as vectors of the tick-borne relapsing fever caused by Borrelia spp. and have also been reported to carry other micro-organisms. The objective of this study was to collect and to identify O. sonrai ticks and to investigate the micro-organisms associated with them. In 2019, an investigation of burrows within human dwellings was conducted in 17 villages in the Niakhar area and in 15 villages in the Sine-Saloum area in the Fatick region of Senegal. Ticks collected from the burrows were identified morphologically and by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Micro-organism screening was performed by bacteria-specific qPCR and some identifications were made by standard PCR and gene sequencing. O. sonrai ticks were found in 100% (17/17) of the villages surveyed in the Niakhar area and in 66% (10/15) of the villages in the Sine-Saloum area. A total of 1275 soft tick specimens were collected from small mammal burrows. The ticks collected were morphologically identified as O. sonrai . About 20% (259/1275) of the specimens were also submitted to MALDI-TOF MS for identification. Among the resulting MS profiles, 87% (139/159) and 95% (95/100) were considered good quality specimens, preserved in alcohol and silica gel, respectively. All spectra of good quality were tested against our MALDI-TOF MS arthropod spectra database and identified as O. sonrai species, corroborating the morphological classification. The carriage of four micro-organisms was detected in the ticks with a high prevalence of Bartonella spp., Anaplasmataceae, and Borrelia spp. of 35, 28, and 26%, respectively, and low carriage of Coxiella burnetii (2%). This study highlights the level of tick infestation in domestic burrows, the inventory of pathogens associated with the O. sonrai tick, and the concern about the potential risk of tick involvement in the transmission of these pathogens in Senegal.

Bed bug has become a major public health pest worldwide. Infestation may result in numerous negative health effects. Homeless shelters are one of the most habitats that can be infested with bed bugs, a few studies have focused on bed bug infestations in these settings. We conducted a survey of infestations of bed bugs in a homeless shelter in southern France, using an innovative seven-level scale (0-6) to assess the degree of infestation, MALDI TOF-MS to identify bed bugs, and a biomolecular tool to detect bacteria. Bed bug infestations were documented in 13% (9/68) of investigated rooms. A total of 184 bed bugs were collected and morphologically identified as Cimex lectularius. MALDI TOF-MS analysis allowed us to obtain high-quality MS spectra for all 184 specimens, to correctly identify all specimens, and included 178/184 (97%) Log Score Values higher than 1.8. Among the bacteria tested, Wolbachia sp. DNA was found in 149/184 (81%) of the bed bugs, and one sample was positive for Coxiella burnetii, the agent of Q fever. Our study is the first of its kind that offers new perspectives for increasing public awareness of the conditions in homeless shelters., (© 2023. The Author(s).)

Hedgehogs are small synanthropic mammals that live in rural areas as well as in urban and suburban areas. They can be reservoirs of several microorganisms, including certain pathogenic agents that cause human and animal public health issues. Hedgehogs are often parasitized by blood-sucking arthropods, mainly hard ticks and fleas, which in turn can also carry various vector-born microorganisms of zoonotic importance. Many biotic factors, such as urbanization and agricultural mechanization, have resulted in the destruction of the hedgehog's natural habitats, leading these animals to take refuge near human dwellings, seeking food and shelter in parks and gardens and exposing humans to zoonotic agents that can be transmitted either directly by them or indirectly by their ectoparasites. In this review, we focus on the microorganisms detected in arthropods sampled from hedgehogs worldwide. Several microorganisms have been reported in ticks collected from these animals, including various Borrelia spp., Anaplasma spp., Ehrlichia spp., and Rickettsia spp. species as well as Coxiella burnetii and Leptospira spp. As for fleas, C. burnetii, Rickettsia spp., Wolbachia spp., Mycobacterium spp. and various Bartonella species have been reported. The detection of these microorganisms in arthropods does not necessarily mean that they can be transmitted to humans and animals. While the vector capacity and competence of fleas and ticks for some of these microorganisms has been proven, in other cases the microorganisms may have simply been ingested with blood taken from an infected host. Further investigations are needed to clarify this issue. As hedgehogs are protected animals, handling them is highly regulated, making it difficult to conduct epidemiological studies on them. Their ectoparasites represent a very interesting source of information on microorganisms circulating in populations of these animals, especially vector-born ones., (© 2023. The Author(s).)

A large outbreak of Monkeypox virus (MPXV) infections has arisen in May 2022 in nonendemic countries. Here, we performed DNA metagenomics using next-generation sequencing with Illumina or Nanopore technologies for clinical samples from MPXV-infected patients diagnosed between June and July 2022. Classification of the MPXV genomes and determination of their mutational patterns were performed using Nextclade. Twenty-five samples from 25 patients were studied. A MPXV genome was obtained for 18 patients, essentially from skin lesions and rectal swabbing. All 18 genomes were classified in clade IIb, lineage B.1, and we identified four B.1 sublineages (B.1.1, B.1.10, B.1.12, B.1.14). We detected a high number of mutations (range, 64-73) relatively to a 2018 Nigerian genome (genome GenBank Accession no. NC&#95;063383.1), which were harbored by a large part of a set of 3184 MPXV genomes of lineage B.1 recovered from GenBank and Nextstrain; and we detected 35 mutations relatively to genome ON563414.3 (a B.1 lineage reference genome). Nonsynonymous mutations occurred in genes encoding central proteins, among which transcription factors and core and envelope proteins, and included two mutations that would truncate a RNA polymerase subunit and a phospholipase d-like protein, suggesting an alternative start codon and gene inactivation, respectively. A large majority (94%) of nucleotide substitutions were G > A or C > U, suggesting the action of human APOBEC3 enzymes. Finally, >1000 reads were identified as from Staphylococcus aureus and Streptococcus pyogenes for 3 and 6 samples, respectively. These findings warrant a close genomic monitoring of MPXV to get a better picture of the genetic micro-evolution and mutational patterns of this virus, and a close clinical monitoring of skin bacterial superinfection in monkeypox patients., (© 2023 Wiley Periodicals LLC.)

Background: To investigate the aetiology of acute undifferentiated fever (AUF) among children under the age of five in Vietnam., Methods: This prospective study was conducted in the Thai Binh paediatric hospital, between July 2020 and July 2021 among children with AUF at admission. Real-time PCR testing 18 microbial pathogens were done on blood samples., Results: 286 children were included, with median age of 16 months. 64.7% were male. 53.9% were positive for at least one pathogen by PCR. Enterovirus, human herpesvirus 6, adenovirus, and varicella zoster virus PCR were positive for 31.1, 12.6, 1.4, and 1.0% patients, respectively. Other pathogens tested negative by PCR. During the hospital stay, based on clinical criteria 47.2% children secondarily presented with signs of respiratory tract infections, 18.9% had hand, foot and mouth disease, 4.6% had chickenpox. 4.2% presented signs of central nervous system infections, 1.0% had dengue (antigenic test) and 1.0% had signs of gastrointestinal infection. Finally, 23.1% patients presented a fever with or without a rash and no other symptoms and ultimately received a diagnosis of AUF., Conclusion: Real-time PCR of blood is useful for detecting pathogens and diagnosing infectious causes of AUF. Further prospective studies with blood and urine culture testing and PCR investigation of not only blood but also cerebrospinal fluid, throat, and skin samples according to symptoms would be of interest to confirm the predominance of viral infections in children with AUF and to guide therapeutic options., (© 2023. The Author(s).)

MALDI-TOF MS has recently been proposed as an accurate tool for arthropod identification, including ticks. In this study, we evaluate and confirm the ability of MALDI-TOF MS, to identify different tick species collected in Cameroon, considering other lines of evidence (morphology and molecular). A total of 1483 adult ticks were collected from cattle in five distinct sites in the Western Highland of Cameroon. Because of engorged status and/or absence of some morphological criteria, some Ixodes spp. and Rhipicephalus spp. were identified to the genus level only. Among those, 944 ticks (543 males and 401 females) were selected for the current work. They were classified into 5 genera and 11 species: Rhipicephalus (Boophilus) microplus (31.7%), Rhipicephalus lunulatus (26%), Amblyomma variegatum (23%), Rhipicephalus sanguineus s.l. (4.8%), of Haemaphysalis leachi group (4.6%), Hyalomma truncatum (2.6%), Hyalomma rufipes (1.7%), Rhipicephalus muhsamae (1.1%), Rhipicephalus (Boophilus) annulatus (0.6%), Rhipicephalus (Boophilus) decoloratus (0.3%), Ixodes rasus (0.1%), Ixodes spp. (0.2%) and Rhipicephalus spp. (3.3%). Tick legs were subjected to MALDI-TOF MS analyzes, and the spectra of 929 (98.4%) specimens were of good quality. Analysis of these spectra provided intra-species reproducibility and interspecies specificity of MS profiles obtained from the different species. Our in-house MALDI-TOF MS arthropod database was upgraded with spectra from 44 specimens of 10 different tick species. Blind testing of good quality spectra revealed that 99% agreed with the morphological identification. Of these, 96.9% had log score values (LSVs) between 1.73 and 2.57. MALDI-TOF MS also allowed to correct the morphological misidentification of 7 ticks, and to identify 32 engorged ticks that were not morphologically identifiable to the species level. This study supports MALDI-TOF MS as a reliable tool for tick identification and provides new data on tick species identification in Cameroon., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Background and Objectives : Hydroxychloroquine (HCQ) combined with azithromycin (AZM) has been widely administered to patients with COVID-19 despite scientific controversies. In particular, the potential of prolong cardiac repolarization when using this combination has been discussed. Materials and Methods : We report a pragmatic and simple safety approach which we implemented among the first patients treated for COVID-19 in our center in early 2020. Treatment contraindications were the presence of severe structural or electrical heart disease, baseline corrected QT interval (QTc) > 500 ms, hypokalemia, or other drugs prolonging QTc that could not be interrupted. Electrocardiogram and QTc was evaluated at admission and re-evaluated after 48 h of the initial prescription. Results : Among the 424 consecutive adult patients (mean age 46.3 ± 16.1 years; 216 women), 21.5% patients were followed in conventional wards and 78.5% in a day-care unit. A total of 11 patients (2.6%) had contraindications to the HCQ-AZ combination. In the remaining 413 treated patients, there were no arrhythmic events in any patient during the 10-day treatment regimen. QTc was slightly but statistically significantly prolonged by 3.75 ± 25.4 ms after 2 days of treatment ( p = 0.003). QTc prolongation was particularly observed in female outpatients <65 years old without cardiovascular disease. Ten patients (2.4%) developed QTc prolongation > 60 ms, and none had QTc > 500 ms. Conclusions : This report does not aim to contribute to knowledge of the efficacy of treating COVID-19 with HCQ-AZ. However, it shows that a simple initial assessment of patient medical history, electrocardiogram (ECG), and kalemia identifies contraindicated patients and enables the safe treatment of COVID-19 patients with HCQ-AZ. QT-prolonging anti-infective drugs can be used safely in acute life-threatening infections, provided that a strict protocol and close collaboration between infectious disease specialists and rhythmologists are applied.

Members of the Cimicidae family are significant pests for mammals and birds, and they have attracted medical and veterinary interest. A number of recent studies have investigated bed bugs, due to their dramatic resurgence all over the world. Indeed, bed bugs are of significant public health and socioeconomic importance since they lead to financial burdens and dermatological complications and may have mental and psychological consequences. It is important to note that certain cimicids with a preference for specific hosts (birds and bats) use humans as an alternative host, and some cimicids have been reported to willingly feed on human blood. In addition, members of the Cimicidae family can lead to economic burdens and certain species are the vectors for pathogens responsible for diseases. Therefore, in this review, we aim to provide an update on the species within the Cimicidae family that have varying medical and veterinary impacts, including their distribution and their associated microorganisms. Various microbes have been documented in bed bugs and certain important pathogens have been experimentally documented to be passively transmitted by bed bugs, although no conclusive evidence has yet associated them with epidemiological outbreaks. Additionally, among the studied cimicids (bat bugs, chicken bugs, and swallow bugs), only the American swallow bug has been considered to be a vector of several arboviruses, although there is no proven evidence of transmission to humans or animals. Further studies are needed to elucidate the reason that certain species in the Cimicidae family cannot be biologically involved in transmission to humans or animals. Additional investigations are also required to better understand the roles of Cimicidae family members in the transmission of human pathogens in the field.

Monitoring of tickborne diseases is critical for prevention and management. We analyzed 418 ticks removed from 359 patients during 2014-2021 in Marseille, France, for identification and bacteria detection. Using morphology, molecular methods, or matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, we identified 197 (47%) Ixodes, 136 (33%) Dermacentor, 67 (16%) Rhipicephalus, 8 (2%) Hyalomma, 6 (1%) Amblyomma, 2 (0.5%) Argas, and 2 (0.5%) Haemaphysalis tick species. We also detected bacterial DNA in 241 (58%) ticks. The most frequent bacterial pathogens were Rickettsia raoultii (17%) and R. slovaca (13%) in Dermacentor ticks, Borrelia spp. (9%) in Ixodes ticks, and R. massiliae (16%) in Rhipicephalus ticks. Among patients who were bitten, 107 had symptoms, and tickborne diseases were diagnosed in 26, including scalp eschar and neck lymphadenopathy after tick bite and Lyme borrelioses. Rapid tick and bacteria identification using a combination of methods can substantially contribute to clinical diagnosis, treatment, and surveillance of tickborne diseases.

In endemic malaria areas, Plasmodium is currently diagnosed mainly through the use of rapid diagnostic tests (RDTs). However, in Senegal, many causes of fever remain unknown. Tick-borne relapsing fever, an often-neglected public health problem, is the main cause of consultation for acute febrile illness after malaria and flu in rural areas. Our objective was to test the feasibility of extracting and amplifying DNA fragments by quantitative polymerase chain reaction (qPCR) from malaria-negative RDTs for Plasmodium falciparum (malaria Neg RDTs P.f) to detect Borrelia spp. and other bacteria. Between January and December 2019, malaria Neg RDTs P.f were collected on a quarterly basis in 12 health facilities in four regions of Senegal. The DNA extracted from the malaria Neg RDTs P.f was tested using qPCR and the results were confirmed by standard PCR and sequencing. Only Borrelia crocidurae DNA was detected in 7.22% (159/2,202) of RDTs. The prevalence of B. crocidurae DNA was higher in July (16.47%, 43/261) and August (11.21%, 50/446). The annual prevalence was 9.2% (47/512) and 5.0% (12/241) in Ngayokhem and Nema-Nding, respectively, health facilities in the Fatick region. Our study confirms that B. crocidurae infection is a frequent cause of fever in Senegal, with a high prevalence of cases in health facilities in the regions of Fatick and Kaffrine. Malaria Neg RDTs P.f are potentially a good source of pathogen sampling for the molecular identification of other causes of fever of unknown origin, even in the most remote areas.

Fleas are obligatory blood-sucking ectoparasites of medical and veterinary importance. The identification of fleas and associated flea-borne microorganisms, therefore, plays an important role in controlling and managing these vectors. Recently, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been reported as an innovative and effective approach to the identification of arthropods, including fleas. This study aims to use this technology to identify ethanol-preserved fleas collected in Vietnam and to use molecular biology to search for microorganisms associated with these fleas. A total of 502 fleas were collected from wild and domestic animals in four provinces in Vietnam. Morphological identification led to the recognition of five flea species, namely Xenopsylla cheopis , Xenopsylla astia , Pulex irritans , Ctenocephalides canis , and Ctenocephalides felis . The cephalothoraxes of 300 individual, randomly selected fleas were tested using MALDI-TOF MS and molecular analysis for the identification and detection of microorganisms. A total of 257/300 (85.7%) of the obtained spectra from the cephalothoraxes of each species were of good enough quality to be used for our analyses. Our laboratory MALDI-TOF MS reference database was upgraded with spectra achieved from five randomly selected fleas for every species of Ctenocephalides canis and Ctenocephalides felis . The remaining spectra were then queried against the upgraded MALDI-TOF MS database, which showed 100% correspondence between morphology and MALDI-TOF MS identification for two flea species ( Ctenocephalides canis and Ctenocephalides felis ). The MS spectra of the remaining species (three P. irritans , five X. astia , and two X. cheopis ) were visually generated low-intensity MS profiles with high background noise that could not be used to update our database. Bartonella and Wolbachia spp. were detected in 300 fleas from Vietnam using PCR and sequencing with primers derived from the gltA gene for Bartonella and the 16S rRNA gene for Wolbachia , including 3 Bartonella clarridgeiae (1%), 3 Bartonella rochalimae (1%), 1 Bartonella coopersplainsensis (0.3%), and 174 Wolbachia spp. endosymbionts (58%).

Background: Respiratory and gastrointestinal symptoms and febrile illness are the most common complaints among ill pilgrims attending the Grand Magal of Touba (GMT) in Senegal., Methods: Patients presenting with respiratory or gastrointestinal symptoms or febrile systemic illnesses were recruited between 2018 and 2021 at a healthcare centre close to Touba. Respiratory, gastrointestinal and blood samples were tested for potential pathogens using qPCR., Results: 538 patients were included. 45.5% of these were female, with a median age of 17 years. Of the 326 samples collected from patients with a cough, 62.8% tested positive for at least one virus, including influenza viruses (33.1%). A high positivity rate of bacterial carriage was observed for Haemophilus influenzae (72.7%), Streptococcus pneumoniae (51.2%) and Moraxella catarrhalis (46.0%). Of the 95 samples collected from patients with diarrhoea, 71.3% were positive, with high rates of bacterial carriage, ranging from 4.2% for Tropheryma whipplei to 45.3% for Entero-pathogenic Escherichia coli. Of the 141 blood samples collected from patients with fever, 31.9% were positive including Plasmodium falciparum (21.3%), Borrelia sp. (5.7%) and dengue virus (5.0%)., Conclusion: This study provides insight into the aetiology of most common infections at the GMT on which to base therapeutic options., Competing Interests: Declaration of competing interest A conflicting interest exists when professional judgement concerning a primary interest (such as patient's welfare or the validity of research) may be influenced by a secondary interest (such as financial gain or personal rivalry). It may arise for the authors when they have financial interest that may influence their interpretation of their results or those of others. Examples of potential conflicts of interest include employment, consultancies, stock ownership, honoraria, paid expert testimony, patent applications/registrations, and grants or other funding., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Background: Urogenital schistosomiasis is a major public health concern in sub-Saharan Africa. In Senegal, the disease is endemic in all regions of the country. Recently, WHO strongly recommended including pre-school children and women of reproductive age during a mass drug administration campaign. It is important to describe the burden of the disease in these group at risk using innovative diagnostic tools. This study aimed to assess the use of real-time PCR in the detection of schistosomiasis cases at the community level in a seasonal transmission area., Methods: A cross-sectional survey was carried out in Niakhar located in the centre of Senegal. Pre-schoolchildren, school-aged children and female adolescents and adults were invited to participate in the study in April 2018. Urine samples were collected and examined using Hemastix reagent strips, filtration technique and real-time PCR. Schistosoma haematobium was detected, identified by targeting the Dra1 gene. The prevalence of urogenital schistosomiasis was determined for each group and the performance of the real-time PCR was compared with the conventional techniques., Results: A total of 428 participants were enrolled in this study including 87 (20.4%) pre-school children (1-5 years), 262 (61.3%) school-aged children between (5-14 years), 17 (3.9%) adolescents (15-17 years) and 62 (14.4%) female adults. The comparison of the diagnostic techniques has shown that the prevalence of urogenital schistosomiasis is higher using molecular technique (34.6%) compared to microscopy (20.3%). The percentage rate of haematuria using Hemastix was 23.1%. School-aged children between 5 and 14 years old were the most affected with 29.0% and 43.1% under microscopy and RT-PCR, respectively. In female participants, microscopic prevalence decreases with age, from 21.4% in school-aged children to 17.6% in adolescents and 9.7% in adults. There was good correlation between the number of eggs per 10 ml and the cycle threshold range., Conclusion: These results show the importance of using molecular tools in the surveillance of schistosomiasis particularly in pre-school children and women of reproductive age., (© 2023. The Author(s).)

Human infestations by bed bugs have upsurged globally in recent decades, including in African countries, where recent reports pointed out an increase in infestation. Sympatric dwelling has been described for two species of bed bug parasitizing humans: Cimex hemipterus (the tropical bed bug) and C. lectularius . Identification of these two species is based on morphological characteristics, and gene sequencing, and may also rely on Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS). The present work aimed to assess whether MALDI-TOF MS was applicable for species level identification of immature stages of Cimex . Arthropods were collected in domestic settings in Nouakchott, Mauritania. Identification used morphological keys and MALDI-TOF MS identification was assessed for immature stages. Quantitative PCR and sequencing assays were used to detect arthropod-associated bacteria in each specimen. A total of 92 arthropods were collected, all morphologically identified as C. hemipterus (32 males, 14 females and 45 immature stages). A total of 35/45 specimens produced good quality MALDI-TOF MS spectra. Analysis allowed species level identification of all immature C. hemipterus after their spectra were entered into our in-house MALDI-TOF MS arthropod spectra database. Molecular screening allowed detection of Wolbachia DNA in each specimen. These results suggested that MALDI-TOF MS is a reliable tool for species level identification of Cimex specimens, including immature specimens. Future studies should assess this approach on larger panels of immature specimens for different Cimex species and focus on the precise staging of their different immature developmental stages.

Lice are host-specific insects. Human lice include Pediculus humanus (body lice) which are known to be vectors of serious human bacterial infectious diseases including epidemic typhus, relapsing fever, trench fever and plague; Pediculus humanus capitis (head lice) that frequently affect children; and Pthirus pubis, commonly known as crab lice. In Africa, human infections transmitted by lice remained poorly known and therefore, underestimated, perhaps due to the lack of diagnostic tools and professional knowledge. In this paper we review current knowledge of the microorganisms identified in human lice in the continent of Africa, in order to alert health professionals to the importance of recognizing the risk of lice-related diseases., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to disclose., (Copyright © 2022. Published by Elsevier B.V.)

Background: Subcutaneous (SC) administration of antibiotics represents an attractive alternative to the intravenous (IV) route., Methods: We performed a systematic electronic search of PubMed and the Cochrane Library for all articles published prior to April 2022, using the key terms and MeSH terms 'subcutaneous', 'antibiotic' and the international non-proprietary name of antibiotics., Results: A total of 30 studies were selected including data on the efficacy and tolerability of antibiotics, and seven studies that were conducted in healthy subjects, for relevant information regarding the safety and tolerability of antibiotics. Comparative studies have shown that efficacy is similar for the SC and IV routes for ceftriaxone, teicoplanin and ertapenem. The SC use of other antibiotics such as ampicillin, ceftazidime, cefepime, piperacillin/tazobactam, metronidazole and fosfomycin has also been described. These results have largely been corroborated by pharmacokinetic/pharmacodynamic analyses, especially for time-dependent antibiotics. Complications of SC treatment are rarely severe, with no reports of bacteraemia or other invasive infection related to this route of administration. Therapeutic drug monitoring has been proposed to adapt the dose and avoid toxicity., Discussion: The rationale for using SC administration of ceftriaxone, ertapenem and teicoplanin is strong in patients with non-severe infections. It is already commonly practised in some countries, particularly in France. Other antibiotics could be administered subcutaneously, but further studies are needed to validate their use in clinical practice. Further research is needed to safely generalize and optimize this route of administration whenever possible. This would reduce the risk of catheter-related infections and their complications, together with the length of hospital stay., (© The Author(s) 2022. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We enrolled 136 patients with laboratory-confirmed monkeypox during June 4-August 31, 2022, at the University Hospital Institute Méditerranée Infection in Marseille, France. The median patient age was 36 years (interquartile range 31-42 years). Of 136 patients, 125 (92%) were men who have sex with men, 15 (11%) reported previous smallpox vaccinations, and 21 (15.5%) were HIV-positive. The most frequent lesion locations were the genitals (68 patients, 53%), perianal region (65 patients, 49%), and oral/perioral area (22 patients, 17%). Lesion locations largely corresponded with the route of contamination. Most (68%) patients had isolated anal, genital, or oral lesions when they were first seen, including 56 (61%) who had >1 positive site without a visible lesion. Concurrent sexually transmitted infections were diagnosed in 19 (15%) patients, and 7 patients (5%) were asymptomatic. We recommend vaccination campaigns, intensified testing for sexually transmitted infections, and increased contact tracing to control the ongoing monkeypox outbreak.

Ornithodoros sonrai ( O. sonrai ) ticks are the only known vectors of Borrelia crocidurae , an agent of tick-borne relapsing fever (TBRF) borreliosis. Rodents serve as principal natural reservoirs for Borrelia . Our research objective was to detect TBRF Borrelia and other zoonotic bacterial infections in ticks and in house mice Mus musculus domesticus , an invasive species currently expanding in rural northern Senegal. Real-time and conventional PCR were utilized for detecting Borrelia and other bacterial taxa. The analyses were performed on 253 specimens of O . sonrai and 150 samples of brain and spleen tissue from rodents. Borrelia crocidurae was found in one O. sonrai tick and 18 Mus musculus domesticus samples, with prevalences of 0.39 percent and 12 percent, respectively, as well as Ehrlichia sp. in one Mus musculus domesticus . Further, we were able to detect the presence of a potentially infectious novel species belonging to the Anaplasmataceae family for the first time in O. sonrai ticks. More attention should be paid to the house mouse and O. sonrai ticks, as they can be potential hosts for novel species of pathogenic bacteria in humans.

Mosquito-borne diseases pose a significant threat to humans in almost every part of the world. Key factors such as global warming, climatic conditions, rapid urbanisation, frequent human relocation, and widespread deforestation significantly increase the number of mosquitoes and mosquito-borne diseases in Vietnam, and elsewhere around the world. In southeast Asia, and notably in Vietnam, national mosquito control programmes contribute to reducing the risk of mosquito-borne disease transmission, however, malaria and dengue remain a threat to public health. The aim of our review is to provide a complete checklist of all Vietnamese mosquitoes that have been recognised, as well as an overview of mosquito-borne diseases in Vietnam. A total of 281 mosquito species of 42 subgenera and 22 genera exist in Vietnam. Of those, Anopheles , Aedes , and Culex are found to be potential vectors for mosquito-borne diseases. Major mosquito-borne diseases in high-incidence areas of Vietnam include malaria, dengue, and Japanese encephalitis. This review may be useful to entomological researchers for future surveys of Vietnamese mosquitoes and to decision-makers responsible for vector control tactics.

We investigated respiratory pathogens among ill Hajj pilgrims from Marseille. We also discuss the potential role of point-of-care (POC) rapid molecular diagnostic tools for this purpose. Clinical data were collected using a standardised questionnaire. Influenza A and B viruses, human rhinovirus and human coronaviruses, Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae and Klebsiella pneumoniae were investigated using real-time PCR in respiratory samples obtained during travel, at the onset of symptoms. 207 participants were included. A cough, expectoration, rhinitis and a sore throat were the most frequent respiratory symptoms, followed by loss of voice and dyspnoea. 38.7% and 25.1% of pilgrims reported a fever and influenza-like symptoms, respectively. 59.4% pilgrims received antibiotics. Rhinovirus (40.6%) was the most frequent pathogen, followed by S. aureus (35.8%) and H. influenzae (30.4%). Virus and bacteria co-infections were identified in 28.5% of participants. 25.1% pilgrims who were positive for respiratory bacteria did not receive antibiotic treatment. In the context of the Hajj pilgrimage, it is important to detect infections that can be easily managed with appropriate treatment, and those that can affect prognosis, requiring hospitalisation. POC rapid molecular diagnostic tools could be used for patient management at small Hajj medical missions and to rationalise antibiotic consumption among Hajj pilgrims.

This study used MALDI-TOF MS and molecular tools to identify tick species infesting camels from Tamanrasset in southern Algeria and to investigate their associated microorganisms. Ninety-one adult ticks were collected from nine camels and were morphologically identified as Hyalomma spp., Hyalomma dromedarii , Hyalomma excavatum , Hyalomma impeltatum and Hyalomma anatolicum . Next, the legs of all ticks were subjected to MALDI-TOF MS, and 88/91 specimens provided good-quality MS spectra. Our homemade MALDI-TOF MS arthropod spectra database was then updated with the new MS spectra of 14 specimens of molecularly confirmed species in this study. The spectra of the remaining tick specimens not included in the MS database were queried against the upgraded database. All 74 specimens were correctly identified by MALDI-TOF MS, with logarithmic score values ranging from 1.701 to 2.507, with median and mean values of 2.199 and 2.172 ± 0.169, respectively. One H. impeltatum and one H. dromedarii (2/91; 2.20%) tested positive by qPCR for Coxiella burnetii , the agent of Q fever. We also report the first detection of an Anaplasma sp. close to A. platys in H. dromedarii in Algeria and a potentially new Ehrlichia sp. in H. impeltatum .

Leishmania infantum is the agent of visceral leishmaniasis in the Mediterranean basin. It is transmitted by sandflies of the subgenus Larroussius. Although Phlebotomus perniciosus is the most important vector in this area, an atypical Ph. perniciosus easily confused with Ph. longicuspis has been observed in North Africa. MALDI-TOF MS, an important tool for vector identification, has recently been applied for the identification of sandflies. Spectral databases presented in the literature, however, include only a limited number of Larroussius species. Our objective was to create an in-house database to identify Mediterranean sandflies and to evaluate the ability of MALDI-TOF MS to discriminate close species or atypical forms within the Larroussius subgenus. Field-caught specimens (n = 94) were identified morphologically as typical Ph. perniciosus (PN; n = 55), atypical Ph. perniciosus (PNA; n = 9), Ph. longicuspis (n = 9), Ph. ariasi (n = 9), Ph. mascittii (n = 3), Ph. neglectus (n = 5), Ph. perfiliewi (n = 1), Ph. similis (n = 9) and Ph. papatasi (n = 2). Identifications were confirmed by sequencing of the mtDNA CytB region and sixteen specimens were included in the in-house database. Blind assessment on 73 specimens (representing 1073 good quality spectra) showed a good agreement (98.5%) between MALDI-TOF MS and molecular identification. Discrepancies concerned confusions between Ph. perfiliewi and Ph. perniciosus. Hierarchical clustering did not allow classification of PN and PNA. The use of machine learning, however, allowed discernment between PN and PNA and between the lcus and lcx haplotypes of Ph. longicuspis (accuracy: 0.8938 with partial-least-square regression and random forest models). MALDI-TOF MS is a promising tool for the rapid and accurate identification of field-caught sandflies. The use of machine learning could allow to discriminate similar species.

Mosquitoes (Diptera: Culicidae ) are of significant public health importance because of their ability to transmit major diseases to humans and animals, and are considered as the world's most deadly arthropods. In recent decades, climate change and globalization have promoted mosquito-borne diseases' (MBDs) geographic expansion to new areas, such as North African countries, where some of these MBDs were unusual or even unknown. In this review, we summarize the latest data on mosquito vector species distribution and MBDs affecting both human and animals in North Africa, in order to better understand the risks associated with the introduction of new invasive mosquito species such as Aedes albopictus . Currently, 26 mosquito species confirmed as pathogen vectors occur in North Africa, including Aedes (five species), Culex (eight species), Culiseta (one species) and Anopheles (12 species). These 26 species are involved in the circulation of seven MBDs in North Africa, including two parasitic infections (malaria and filariasis) and five viral infections (WNV, RVF, DENV, SINV and USUV). No bacterial diseases have been reported so far in this area. This review may guide research studies to fill the data gaps, as well as helping with developing effective vector surveillance and controlling strategies by concerned institutions in different involved countries, leading to cooperative and coordinate vector control measures.

This study aimed to detect and identify microorganisms in ticks collected in the Western Highlands of Cameroon. Quantitative real-time and standard PCR assays, coupled with sequencing, were used. A total of 944 ticks collected from cattle in five distinct sites in Cameroon were selected for the analyses. They belonged to five genera ( Amblyomma , Hyalomma , Rhipicephalus , Haemaphysalis, and Ixodes ) and twelve species. Real-time PCR revealed that 23% ( n = 218) of the ticks were positive for Rickettsia spp., 15% ( n = 141) for bacteria of the Anaplasmataceae family, 3% ( n = 29) for Piroplasmida, 0.5% ( n = 5) for Coxiella burnetii, 0.4% ( n = 4) for Borrelia spp., and 0.2% ( n = 2) for Bartonella spp. The co-infection rate (3.4%, n = 32) involved mainly Rickettsia spp. and Anaplasmataceae. Of the Rickettsia spp. positive ticks, the targeted PCR and sequencing yielded Rickettsia africae (78.9%) , Rickettsia aeschlimannii (6.4%), Rickettsia massiliae (7.8%), Candidatus Rickettsia barbariae (0.9%), and Rickettsia sp. (0.9%). Anaplasmataceae included Anaplasma marginale (4.3%), Anaplasma platys (1.4%), Anaplasma centrale (0.7%), Ehrlichia ruminantium (0.7%), Wolbachia sp., Candidatus Ehrlichia rustica (13.5%), Candidatus Ehrlichia urmitei (7%), and an uncultured Ehrlichia sp. (4.2%). Borrelia theileri was identified in one Rhipicephalus microplus tick. Unfortunately, Piroplasmida could not be identified to the species level. This study demonstrates that in Cameroon, ticks harbour a wide variety of microorganisms and present a risk of zoonotic diseases.

Background: Respiratory and gastrointestinal symptoms are frequent in pilgrims at the Grand Magal of Touba (GMT)., Methods: Pilgrims were prospectively investigated in 2017-2021 for demographics, chronic conditions, preventive measures, respiratory and gastrointestinal symptoms, and pathogen carriage using PCR assays., Results: 535 pilgrims were included. 54.8% and 13.3% reported respiratory and gastrointestinal symptoms, respectively. 18.4% acquired respiratory viruses, notably rhinovirus (10.1%) and coronaviruses (5.6%) and 39.9% bacteria, notably Haemophilus influenzae (18.9%) and Streptococcus pneumoniae (14.1%). The acquisition of gastrointestinal pathogens was lower, with enteroaggregative Escherichia coli (18.9%) and enteropathogenic Escherichia coli (10.5%) being the most frequent. A decrease was observed in the acquisition rates of pathogens in 2020-2021 GMT. Female pilgrims were more at risk of respiratory and gastrointestinal symptoms. Respiratory symptoms were associated with virus acquisition (aRR: 2.20, 95%CI [1.38-3.50]) and S. pneumoniae acquisition (aRR: 2.76, 95%CI = [1.64-4.62]). Using hand soap was associated with a decrease in the acquisition of rhinovirus (aRR: 0.42, 95%CI [0.22-0.80]) and coronavirus (aRR: 0.42, 95%CI [0.22-0.81]). Using face masks was associated with a decrease in reporting of respiratory symptoms (aRR: 0.54, 95% [0.35-0.86])., Conclusion: Hand washing with soap and wearing face masks should be recommended to GMT pilgrims., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Background: The control of snails intermediate hosts remains an effective strategy to limit schistosomiasis transmission despite the widespread mass de-worming campaign based on praziquantel. Therefore, the study of snail biology could help to improve snails control strategies. This study evaluated the development of Biomphalaria pfeifferi , Bulinus senegalensis and Bulinus truncatus in various water sources and their resistance to salinity., Methods: Five day-old juveniles individuals issued from adult snails non-shedding Schistosoma spp cercariae were breed in distilled water, commercial mineral water, ground pump water and well water. Snail's survival rate and size were measured over a period of 40 days. These two parameters were also measured over 30 days in increasing saline solutions (1 g/l, 3.5 g/l and 4 g/l) made of well water to which sodium chloride was added., Results: B. truncatus growth was not hampered by any water sources with a survival rate between 68% and 84% (log rank X 2 = 1.86, df = 3, p = 0.60). Despite a poor survival rate (8%) in distilled water, B. pfeifferi , has adapted to other water sources with a survival fraction between 88% and 96% (log rank X 2 = 61.94, df = 3, p < 0.0001). B. senegalensis development was very delicate with low survival rate of 4% in distilled water, 20% in well water and 24% in commercial mineral water and ground pump water (log rank X 2 = 13.24, df = 3, p = 0,004). For each species, even if the difference is not significant, the size of snails is larger with well water and pump water compared to distilled and commercial mineral water. B. pfeifferi survival rate was at 45% in both three saline solutions at day 30. B. senegalensis population collapsed at day 10 in 4 g/l saline solution and persisted until day 30 in both 1.5 g/l and 3.5 g/l solution. B. truncatus also persisted with a survival rate at 20% in 1.5 g/l but collapsed at day 5 and 15 in 4 g/l and 3.5 g/l solution, respectively., Conclusion: The differences in adaptation between snails species show the need to take into account the water sources for snail breeding in the laboratory. Further studies could help to determine the optimal water quality for each snail species in order to standardize breeding conditions. This study could contribute to the understanding of the dynamics and distribution of snails in natural conditions., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

Objective of this study is to find the optimal conditions for preparing the samples, resulting in quality, reproducible and specific MS spectra of the ticks, with a shelf life in 70% ethanol of more than ten years. Amblyomma (Am.) variegatum species which had been stored in alcohol for more than twenty years and for which numerous specimens were available were used to compare the performance of four protocols tested. Spectra of insufficient quality were obtained from Am. variegatum legs preserved in alcohol for long periods with the reference protocol, named DO that we had set up years ago. The same observation was made on the spectra from Am. variegatum legs from dry (evaporated alcohol, DO-mod protocol). With new protocols named ReDO and PReDO the spectra were of good quality with high intensities (> 3000 a.u.). Blind testing showed that 94%, and 93% of the spectra were correctly identified with relevant log score values (LSVs ≥1.8), respectively for ReDO and PReDO protocols. All soft ticks treated in this study by PReDO protocol exhibited low intensity spectra with background noise. This study revealed that MALDI-TOF MS is able to identify hard ticks stored during decades in alcohol or dry (evaporated alcohol). SIGNIFICANCE OF THE STUDY: The correct identification of ticks, including vectors responsible for the transmission of infectious diseases in humans and animals is essential for their control. MALDI-TOF MS, a proteomic tool that has emerged in recent years, has become an innovative, accurate and alternative tool for the identification of arthropods, including ticks. However, previous studies reported that preservation of arthropods in alcohol modified the MS spectra obtained from specimens of the same species freshly collected or frozenly stored. In this study, a standard protocol was established for the identification of tick collections which had been stored for more than ten years in alcohol. Four different protocols were assessed. The analysis of the results showed that among the four protocols tested, two protocols named ReDO (Rehydration and incubation of the legs in 40 μl of HPLC water for 12 h in a dry bath at 37°) and PreDO (Drying of the legs for 12 h in a dry bath at 37 °C followed by rehydration and incubation in 40 μl of HPLC water for 12 h.) seem to be more appropriate for the MALDI-TOF MS identification of ticks from old collections preserved in alcohol or dry. This study is promising for the future, as it will make it possible to create a MALDI-TOF MS database from a wide range of ticks which have been stored for a long time in alcohol or which are dry stored in laboratories and museums around the world., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background: Prognosis of herpetic encephalitis remains severe, with a high proportion of deaths and sequelae. Its treatment is based on acyclovir, but the precise and most effective modalities of this treatment are not established. The objective of this study was to determine them., Methods: For this, we carried out a descriptive, retrospective, monocentric study, using the current coding database at Marseille University Hospitals. Cohort was intended to be exhaustive for the disease, from January 2000 to June 2019, including patients hospitalized in intensive care and conventional hospitalization sector. Patients (n = 76) included were at least 16 years of age and had a clinical presentation, cerebral Magnetic Resonance Imaging, and/or electroencephalogram abnormalities consistent with herpetic encephalitis confirmed by a positive HSV-PCR in the CSF. Clinical data and treatment, including the doses actually administered to the patient, were compared according to patient's outcome., Results: The mortality rate was 12%, whereas 49% had complete recovery and 39% sequelae impeding independence. Poor outcome was statistically associated with persistence of confusion, aphasia, and impaired consciousness lasting more than 5 days, superinfection, status epilepticus, and length of stay in intensive care unit. A statistical decision tree, constructed using the Classification And Regression Tree model, to prioritize treatment management, showed two main factors that influence the outcome: the patient's weight, and the average daily acyclovir dose actually administered., Conclusion: These results suggest to modify acyclovir management in herpetic encephalitis, for low-weight patients (< 79 kg) with a minimum dosage of 2550 mg/day (850 mg/ 8 h), when possible., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.)

Background: In the last decade, an innovative approach has emerged for arthropod identification based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Increasing interest in applying the original technique for arthropod identification has led to the development of a variety of procedures for sample preparation and selection of body parts, among others. However, the absence of a consensual strategy hampers direct inter-study comparisons. Moreover, these different procedures are confusing to new users. Establishing optimized procedures and standardized protocols for mosquito identification by MALDI-TOF MS is therefore a necessity, and would notably enable the sharing of reference MS databases. Here, we assess the optimal conditions for mosquito identification using MALDI-TOF MS profiling., Methods: Three homogenization methods, two of which were manual and one automatic, were used on three distinct body parts (legs, thorax, head) of two mosquito laboratory strains, Anopheles coluzzii and Aedes aegypti, and the results evaluated. The reproducibility of MS profiles, identification rate with relevant scores and the suitability of procedures for high-throughput analyses were the main criteria for establishing optimized guidelines. Additionally, the consequences of blood-feeding and geographical origin were evaluated using both laboratory strains and field-collected mosquitoes., Results: Relevant score values for mosquito identification were obtained for all the three body parts assayed using MALDI-TOF MS profiling; however, the thorax and legs were the most suitable specimens, independently of homogenization method or species. Although the manual homogenization methods were associated with a high rate of identification on the three body parts, this homogenization mode is not adaptable to the processing of a large number of samples. Therefore, the automatic homogenization procedure was selected as the reference homogenization method. Blood-feeding status did not hamper the identification of mosquito species, despite the presence of MS peaks from original blood in the MS profiles of the three body parts tested from both species. Finally, a significant improvement in identification scores was obtained for field-collected specimens when MS spectra of species from the same geographical area were added to the database., Conclusion: The results of the current study establish guidelines for the selection of mosquito anatomic parts and modality of sample preparation (e.g. homogenization) for future specimen identification by MALDI-TOF MS profiling. These standardized operational protocols could be used as references for creating an international MS database., (© 2022. The Author(s).)

Objectives: We evaluated the 6-week mortality of SARS-CoV-2 hospitalized patients treated using a standardized protocol in 2020 in Marseille, France., Methods: A retrospective monocentric cohort study was conducted in the standard hospital wards at the Institut Hospitalo-Universitaire Méditerranée Infection, between March and December 2020 in adults with SARS-CoV-2 PCR-proven infection., Results: Of the 2111 hospitalized patients (median age, 67 [IQR 55-79] years; 1154 [54.7%] men), 271 were transferred to the intensive care unit (12.8%) and 239 died (11.3%; the mean age of patients who died was 81.2 (±9.9)). Treatment with hydroxychloroquine plus azithromycin (HCQ-AZ), used in 1270 patients, was an independent protective factor against death (0.68 [0.52 - 0.88]). This effect was consistent for all subgroups of age, comorbidities, severity of the disease and comedications with zinc or corticosteroids. Zinc was independently protective against death (0.39 [0.23 - 0.67]), in a subgroup analysis of patients treated with HCQ-AZ without dexamethasone. The use of high-flow oxygen therapy in elderly patients who were not eligible for intensive care unit transfer saved 19 patients (33.9%)., Conclusions: In our 2020 cohort, treating COVID-19 with HCQ-AZ was associated with lower mortality. These results need to be analyzed in the context of academic discussions about observational studies versus randomized clinical trials. More data will deserve to be analyzed in the SARS-Cov 2 variants, vaccination and post-vaccination era., Competing Interests: Prof. Dr Didier Raoult reports personal fees from Scientific board member of Eurofins company, Founder and shareholder of a microbial culture company (Culture Top), received personal fees from Hitachi High-Technologies Corporation, Tokyo, Japan, from 2018 to 2020, Founder and shareholder of Biotechnology “Techno-jouvence”, Founder and shareholder of a Biotech company “Gene and Green TK, Founder and shareholder of rapid diagnosis of infectious diseases company “Pocramé”, outside the submitted work. The authors declare no other competing interests. Funding sources had no role in the design and conduct of the study; collection, management, analysis, and interpretation of the data; and preparation, review, or approval of the manuscript., (© 2022 Lagier et al.)

Bed bugs are known to carry several microorganisms. The purpose of this study was to assess the prevalence of bed bug infestation in two rural areas of Senegal and determine the species present in the population. A screening was conducted to detect some arthropod associated pathogenic bacteria in bed bugs and to evaluate the prevalence of endosymbiont carriage. One survey took place in 17 villages in Niakhar and two surveys in Dielmo and Ndiop and surroundings area in the same 20 villages. Bed bugs collected were identified morphologically and by MALDI-TOF MS tools. Microorganisms screening was performed by qPCR and confirmed by sequencing. During the survey in the Niakhar region, only one household 1/255 (0.4%) in the village of Ngayokhem was found infested by bed bugs. In a monitoring survey of the surroundings of Dielmo and Ndiop area, high prevalence was found during the two rounds of surveys in 65/314 (21%) in 16/20 villages (January-March) and 93/351 (26%) in 19/20 villages (December). All bed bugs were morphologically identified as the species Cimex hemipterus, of which 285/1,637 (17%) were randomly selected for MALDI-TOF MS analysis and bacteria screening. Among the Bacteria tested only Wolbachia (Alphaproteobacteria, Rickettsiales, Rickettsiaceae) DNA was found in 248/276 (90%) of the bedbugs. We briefly describe a high level of non-generalized bed bug infestation in rural Senegal and the diversity of Wolbachia strains carried by C. hemipterus. This study opens perspectives for raising household awareness of bed bug infestations and possibilities for appropriate control., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Persistent fever, defined as fever lasting for 7 days or more at first medical evaluation, has been hardly investigated as a separate clinical entity in the tropics. This study aimed at exploring the frequencies and diagnostic predictors of the ubiquitous priority (i.e., severe and treatable) infections causing persistent fever in the tropics., Methods: In six different health settings across four countries in Africa and Asia (Sudan, Democratic Republic of Congo [DRC], Nepal, and Cambodia), consecutive patients aged 5 years or older with persistent fever were prospectively recruited from January 2013 to October 2014. Participants underwent a reference diagnostic workup targeting a pre-established list of 12 epidemiologically relevant priority infections (i.e., malaria, tuberculosis, HIV, enteric fever, leptospirosis, rickettsiosis, brucellosis, melioidosis, relapsing fever, visceral leishmaniasis, human African trypanosomiasis, amebic liver abscess). The likelihood ratios (LRs) of clinical and basic laboratory features were determined by pooling all cases of each identified ubiquitous infection (i.e., found in all countries). In addition, we assessed the diagnostic accuracy of five antibody-based rapid diagnostic tests (RDTs): Typhidot Rapid IgM, Test-it TM Typhoid IgM Lateral Flow Assay, and SD Bioline Salmonella typhi IgG/IgM for Salmonella Typhi infection, and Test-it TM Leptospira IgM Lateral Flow Assay and SD Bioline Leptospira IgG/IgM for leptospirosis., Results: A total of 1922 patients (median age: 35 years; female: 51%) were enrolled (Sudan, n = 667; DRC, n = 300; Nepal, n = 577; Cambodia, n = 378). Ubiquitous priority infections were diagnosed in 452 (23.5%) participants and included malaria 8.0% (n = 154), tuberculosis 6.7% (n = 129), leptospirosis 4.0% (n = 77), rickettsiosis 2.3% (n = 44), enteric fever 1.8% (n = 34), and new HIV diagnosis 0.7% (n = 14). The other priority infections were limited to one or two countries. The only features with a positive LR ≥ 3 were diarrhea for enteric fever and elevated alanine aminotransferase level for enteric fever and rickettsiosis. Sensitivities ranged from 29 to 67% for the three RDTs targeting S. Typhi and were 9% and 16% for the two RDTs targeting leptospirosis. Specificities ranged from 86 to 99% for S. Typhi detecting RDTs and were 96% and 97% for leptospirosis RDTs., Conclusions: Leptospirosis, rickettsiosis, and enteric fever accounted each for a substantial proportion of the persistent fever caseload across all tropical areas, in addition to malaria, tuberculosis, and HIV. Very few discriminative features were however identified, and RDTs for leptospirosis and Salmonella Typhi infection performed poorly. Improved field diagnostics are urgently needed for these challenging infections., Trial Registration: NCT01766830 at ClinicalTrials.gov., (© 2022. The Author(s).)