The animal reservoir of SARS-CoV-2 is unknown despite reports of SARS-CoV-2-related viruses in Asian Rhinolophus bats
, including the closest virus from R. affinis, RaTG13 (refs.
), and pangolins
. ...SARS-CoV-2 has a mosaic genome, to which different progenitors contribute. The spike sequence determines the binding affinity and accessibility of its receptor-binding domain to the cellular angiotensin-converting enzyme 2 (ACE2) receptor and is responsible for host range
. SARS-CoV-2 progenitor bat viruses genetically close to SARS-CoV-2 and able to enter human cells through a human ACE2 (hACE2) pathway have not yet been identified, although they would be key in understanding the origin of the epidemic. Here we show that such viruses circulate in cave bats living in the limestone karstic terrain in northern Laos, in the Indochinese peninsula. We found that the receptor-binding domains of these viruses differ from that of SARS-CoV-2 by only one or two residues at the interface with ACE2, bind more efficiently to the hACE2 protein than that of the SARS-CoV-2 strain isolated in Wuhan from early human cases, and mediate hACE2-dependent entry and replication in human cells, which is inhibited by antibodies that neutralize SARS-CoV-2. None of these bat viruses contains a furin cleavage site in the spike protein. Our findings therefore indicate that bat-borne SARS-CoV-2-like viruses that are potentially infectious for humans circulate in Rhinolophus spp. in the Indochinese peninsula.
I attempt in this essay to shed new light on the origins of Louis Pasteur's uniquely progressive mind and spirit and the various factors in his background and upbringing that shaped them. There is, ...however, very limited documentation on the early period of Pasteur's life, apart from his son-in-law's (René Vallery-Radot) biographical account, and this is considered by many as more akin to a work of hagiography. We do have, on the other hand, Pasteur's correspondence with his parents and sisters as compiled and annotated by his grandson, Louis Pasteur Vallery-Radot. Using these limited sources, combined with what we know about Pasteur's home environment and early education, his cultural influences (like the books he read), and his drawings and etchings, I have attempted in this essay to hypothesize regarding the influences during Pasteur's childhood, adolescence, and very early adult years as a scientist and how they contributed to the formation of Pasteur's mind and spirit, while fully acknowledging the extent to which this is based on indirect evidence and, occasionally, outright speculation.
Estimating the exposure of individuals to mosquito-borne diseases is a key measure used to evaluate the success of vector control operations. The gold standard is to use human landing catches where ...mosquitoes are collected off the exposed limbs of human collectors. This is however an unsatisfactory method since it potentially exposes individuals to a range of mosquito-borne diseases. In this study several sampling methods were compared to find a method that is representative of the human-biting rate outdoors, but which does not expose collectors to mosquito-borne infections. The sampling efficiency of four odour-baited traps were compared outdoors in rural Lao PDR; the human-baited double net (HDN) trap, CDC light trap, BG sentinel trap and Suna trap. Subsequently the HDN, the best performing trap, was compared directly with human landing catches (HLC), the 'gold standard', for estimating human-biting rates. HDNs collected 11-44 times more mosquitoes than the other traps, with the exception of the HLC. The HDN collected similar numbers of Anopheles (Rate Ratio, RR = 1.16, 95% Confidence Intervals, 95% CI = 0.61-2.20) and Culex mosquitoes (RR = 1.26, 95% CI = 0.74-2.17) as HLC, but under-estimated the numbers of Aedes albopictus (RR = 0.45, 95% CI = 0.27-0.77). Simpson's index of diversity was 0.845 (95% CI 0.836-0.854) for the HDN trap and 0.778 (95% CI 0.769-0.787) for HLC, indicating that the HDN collected a greater diversity of mosquito species than HLC. Both HLC and HDN can distinguish between low and high biting rates and are crude ways to measure human-biting rate. The HDN is a simple and cheap method to estimate the human-biting rate outdoors without exposing collectors to mosquito bites.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The mosquito Aedes aegypti is the primary vector of several arboviruses, such as dengue, chikungunya and Zika, and represents a major public health problem in Southeast Asia. In Laos, where dengue is ...reemerging, several Ae. aegypti populations from the capital Vientiane have shown resistance to the organophosphate temephos, a commonly-used larvicide for public health interventions.
Here, we tested the insecticide susceptibility of a wild larval population of Ae. aegypti against Bacillus thuringiensis israelensis (Bti), diflubenzuron, pyriproxyfen and spinosad. Residual efficacies of Bti (VectobacWG®), diflubenzuron (Killmos®) and temephos (Abate®) were then evaluated under simulated field conditions against the wild Ae. aegypti population.
The larval bioassays showed that the wild Ae. aegypti strain was moderately resistant to temephos and spinosad (resistance ratio, RR < 5) and fully susceptible to the other insecticides (RR = 1). The simulated field trial bioassays showed that all of the insecticides tested remained above the WHO acceptable larvicide threshold after 28 weeks.
These results suggest that Bti and diflubenzuron may be promising alternative larvicides for controlling dengue vectors in water-storage containers in Laos, especially against Ae. aegypti populations, in which resistance to temephos has been detected.
The first documented chikungunya virus (CHIKV) outbreak in Lao People's Democratic Republic (Lao PDR) occurred in 2012-2013. Since then, several imported and a few autochthonous cases were identified ...by the national arbovirus surveillance network. The present study aimed to summarize the main genetic features of the CHIKV strains detected in Lao PDR between 2014 and 2020. Samples from Lao patients presenting symptoms compatible with a CHIKV infection were centralized in Vientiane Capital city for real-time RT-PCR screening. Molecular epidemiology was performed by sequencing the E2-6K-E1 region. From 2014 to 2020, two Asian lineage isolates (e.g. French Polynesia; Indonesia), one ECSA-IOL lineage isolate (e.g. Thailand) and one unclassified (e.g. Myanmar) were imported in Vientiane Capital city. Sequences from the autochthonous cases recorded in the Central and Southern parts of the country between July and September 2020 belonged to the ECSA-IOL lineage and clustered with CHIKV strains recently detected in neighboring countries. These results demonstrate the multiple CHIKV introductions in Lao PDR since 2014 and provide evidence for sporadic and time-limited circulation of CHIKV in the country. Even if the circulation of CHIKV seems to be geographically and temporally limited in Lao PDR, the development of international tourism and trade may cause future outbreaks of CHIKV in the country and at the regional level.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Since Metchnikoff developed his views on the intestinal microflora, much effort has been devoted to understanding the role of gut microbiomes in metazoan physiology. Despite impressive data sets that ...have been generated by associating a phenotype-causing commensal community with its corresponding host phenotype, the field continues to suffer from descriptive and often contradictory reports. Hence, we cannot yet draw clear conclusions as to how the modifications of microbiomes cause physiological changes in metazoans. Unbiased, large-scale genetic screens to identify key genes, on both microbial and host sides, will be essential to gain mechanistic insights into gut-microbe interactions. The Drosophila genome-commensal microbiome genetic model has proven to be well suited to dissect the complex reciprocal cross talk between the host and its microbiota. In this review, we present a historical account, current views, and novel perspectives for future research directions based on the insights gleaned from the Drosophila gut-microbe interaction model.
The yellow fever mosquito Aedes aegypti is the major vector of dengue, yellow fever, Zika, and Chikungunya viruses. Worldwide vector control is largely based on insecticide treatments but, ...unfortunately, vector control programs are facing operational challenges due to mosquitoes becoming resistant to commonly used insecticides. In Southeast Asia, resistance of Ae. aegypti to chemical insecticides has been documented in several countries but no data regarding insecticide resistance has been reported in Laos. To fill this gap, we assessed the insecticide resistance of 11 Ae. aegypti populations to larvicides and adulticides used in public health operations in the country. We also investigated the underlying molecular mechanisms associated with resistance, including target site mutations and detoxification enzymes putatively involved in metabolic resistance.
Bioassays on adults and larvae collected in five provinces revealed various levels of resistance to organophosphates (malathion and temephos), organochlorine (DDT) and pyrethroids (permethrin and deltamethrin). Synergist bioassays showed a significant increased susceptibility of mosquitoes to insecticides after exposure to detoxification enzyme inhibitors. Biochemical assays confirmed these results by showing significant elevated activities of cytochrome P450 monooxygenases (P450), glutathione S-transferases (GST) and carboxylesterases (CCE) in adults. Two kdr mutations, V1016G and F1534C, were detected by qPCR at low and high frequency, respectively, in all populations tested. A significant negative association between the two kdr mutations was detected. No significant association between kdr mutations frequency (for both 1534C and 1016G) and survival rate to DDT or permethrin (P > 0.05) was detected. Gene Copy Number Variations (CNV) were detected for particular detoxification enzymes. At the population level, the presence of CNV affecting the carboxylesterase CCEAE3A and the two cytochrome P450 CYP6BB2 and CYP6P12 were significantly correlated to insecticide resistance.
These results suggest that both kdr mutations and metabolic resistance mechanisms are present in Laos but their impact on phenotypic resistance may differ in proportion at the population or individual level. Molecular analyses suggest that CNV affecting CCEAE3A previously associated with temephos resistance is also associated with malathion resistance while CNV affecting CYP6BB2 and CYP6P12 are associated with pyrethroid and possibly DDT resistance. The presence of high levels of insecticide resistance in the main arbovirus vector in Laos is worrying and may have important implications for dengue vector control in the country.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Construction of the Nam Theun 2 hydroelectric project and flooding of a 450 km(2) area of mountain plateau in south-central Lao PDR resulted in the resettlement of 6,300 people to newly built homes. ...We examined whether new houses would have altered risk of house entry by mosquitoes compared with traditional homes built from poorer construction materials.
Surveys were carried out in the Nam Theun 2 resettlement area and a nearby traditional rice farming area in 2010. Mosquitoes were sampled in bedrooms using CDC light traps in 96 resettlement houses and 96 traditional houses and potential risk factors for mosquito house entry were recorded. Risk of mosquito house entry was more than twice as high in traditional bamboo houses compared with those newly constructed from wood (Putative Japanese Encephalitis (JE) vector incidence rate ratio (IRR) = 2.26, 95% CI 1.38-3.70, P = 0.001; Anopheline IRR = 2.35, 95% CI: 1.30-4.23, P = 0.005). Anophelines were more common in homes with cattle compared against those without (IRR = 2.32, 95% CI: 1.29-4.17, P = 0.005).Wood smoke from cooking fires located under the house or indoors was found to be protective against house entry by both groups of mosquito, compared with cooking in a separate room beside the house (Putative JE vector IRR = 0.43, 95% CI: 0.26-0.73, P = 0.002; Anopheline IRR = 0.22, 95% CI: 0.10-0.51, P<0.001).
Construction of modern wooden homes should help reduce human-mosquito contact in the Lao PDR. Reduced mosquito contact rates could lead to reduced transmission of diseases such as JE and malaria. Cattle ownership was associated with increased anopheline house entry, so zooprophylaxis for malaria control is not recommended in this area. Whilst wood smoke was protective against putative JE vector and anopheline house entry we do not recommend indoor cooking since smoke inhalation can enhance respiratory disease.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Although the malaria burden in the Lao PDR has gradually decreased, the elimination of malaria by 2030 presents many challenges. Microscopy and malaria rapid diagnostic tests (RDTs) are used to ...diagnose malaria in the Lao PDR; however, some studies have reported the prevalence of sub-microscopic Plasmodium infections or asymptomatic Plasmodium carriers in endemic areas. Thus, highly sensitive detection methods are needed to understand the precise malaria situation in these areas.
A cross-sectional malaria field survey was conducted in 3 highly endemic malaria districts (Xaysetha, Sanamxay, Phouvong) in Attapeu province, Lao PDR in 2015, to investigate the precise malaria endemicity in the area; 719 volunteers from these villages participated in the survey. Microscopy, RDTs and a real-time nested PCR were used to detect Plasmodium infections and their results were compared. A questionnaire survey of all participants was also conducted to estimate risk factors of Plasmodium infection. Numbers of infections detected by the three methods were microscopy: P. falciparum (n = 1), P. vivax (n = 2); RDTs: P. falciparum (n = 2), P. vivax (n = 3); PCR: Plasmodium (n = 47; P. falciparum n = 4, P. vivax n = 41, mixed infection n = 2; 6.5%, 47/719). Using PCR as a reference, the sensitivity and specificity of microscopy were 33.3% and 100.0%, respectively, for detecting P. falciparum infection, and 7.0% and 100.0%, for detecting P. vivax infection. Among the 47 participants with parasitemia, only one had a fever (≥37.5°C) and 31 (66.0%) were adult males. Risk factors of Plasmodium infection were males and soldiers, whereas a risk factor of asymptomatic Plasmodium infection was a history of ≥3 malaria episodes.
There were many asymptomatic Plasmodium carriers in the study areas of Attapeu province in 2015. Adult males, probably soldiers, were at high risk for malaria infection. P. vivax, the dominant species, accounted for 87.2% of the Plasmodium infections among the participants. To achieve malaria elimination in the Lao PDR, highly sensitive diagnostic tests, including PCR-based diagnostic methods should be used, and plans targeting high-risk populations and elimination of P. vivax should be designed and implemented.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
...the PCR amplicons were purified with Performa DTR Gel Filtration Cartridges (Edge Bio, United States of America) and sequenced by an ABI Genetic Analyzer model 3130XL (Life Technologies, Japan). ...Furthermore, a recent study reported that 63% (28/44) of long-tailed macaques in Laos were infected with P. cynomolgi, which can also be infectious to humans 8,10. ...an investigation of human infection with P. cynomolgi is also needed in Laos. ...previous studies demonstrated that false-positive results for P. vivax and P. falciparum have been observed for P. knowlesi mono-infection with certain RDTs 11,12. First case of a naturally acquired human infection with Plasmodium cynomolgi.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
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