Vector Exposure Research Articles

Presumptive West Nile Virus infection in a Komodo Dragon (Varanus komodoensis)

Abstract A 3-yr-old, male Komodo dragon (Varanus komodoensis) presented acutely in early October 2021 with an abnormal posture and asymmetric tetraparesis. The subadult lizard had spent the previous 4 months either in an outdoor space or in an indoor space with open windows. Physical examination, radiographs, and complete blood count and chemistry panel failed to reveal the cause of the neurologic signs. West Nile virus (WNV) plaque reduction serum neutralization on presentation was negative (1:20), as was WNV polymerase chain reaction (PCR) testing and viral isolation on whole blood. Empirical treatment with a daily corticosteroid, course of antibiotics, and encouragement of mild exercise was initiated. Magnetic resonance imaging of the brain and spinal cord was unremarkable, and a lumbar cerebrospinal fluid (CSF) tap revealed a mononuclear pleocytosis and protein elevation. Follow-up WNV serology revealed rising titer and seroconversion 13 days after presentation (1:80) and 32 days after presentation (1:640); however, WNV PCR testing of CSF fluid was negative. Thi Komodo dragon gradually improved over six weeks before being put back on public display 74 days after presentation. The WNV titer remained high (1:640) through the winter of 2021-22, and an anamnestic response was observed following a vaccination series with a killed equine vaccine in the spring of 2022. There is a previous brief report of WNV infection in a captive crocodile monitor (Varanus salvadorii), and an unpublished case in the same species at the first author’s institution. This is the first report of neurologic disease from presumptive WNV infection in a Komodo dragon. We recommend that WNV infection be a differential diagnosis if a varanid lizard present with acute neurologic signs with a history of arthropod vector exposure, particularly in the summer or fall.

Malaria burden and residual transmission: two thirds of mosquito bites may not be preventable with current vector control tools on Bioko Island, Equatorial Guinea

ObjectivesThis study assesses exposure to malaria vector mosquitos that is nonpreventable through use of nets, the contribution of outdoor and indoor biting towards residual vector exposure, and the risk factors for being bitten and for being infected with malaria parasites on Bioko Island, Equatorial Guinea. MethodsHuman behavior and malaria infection data were collected from 13,735 randomly selected residents during cross-sectional surveys, concomitantly with entomological human landing catches, indoors and outdoors, in 20 locations on the Island. Self-reported time of going indoors, going to bed and whether using a net were analyzed to impute for each respondent the number of bites received outdoors and indoors during the night before the survey. ResultsOn average, each person received 2.7 (95% CI: 2.6-2.8) bites per night outdoors, 8.5 (8.3 to 8.7) bites indoors if not using a net, and 4.7 (4.5 to 4.8) bites indoors if using a net. Malaria infection was associated with more bites, regardless of whether received indoors or outdoors. Older age, male gender, not using a net, rural location, and going indoors later increased the risk of being bitten. The proportion of bites not averted by using a net was estimated as 66% (61 to 71). ConclusionsA large proportion of biting, mostly indoors, may not be preventable by bednets. Tools targeting indoor biting should be prioritized in Bioko. Novel vector control tools are urgently needed to reduce overall exposure to mosquito bites.

Comparative Analysis of Phlebotomus argentipes Vector of Leishmaniasis in India and Sri Lanka.

Phlebotomus argentipes is the predominant sandfly vector of leishmaniasis in the Indian subcontinent. India and Sri Lanka primarily report visceral and cutaneous leishmaniasis caused by Leishmania donovani. We compared Ph. argentipes from two locations, focusing on its morphological, molecular, and salivary protein characteristics. Sandflies were captured using CDC light traps and cattle-baited net traps. Species identification and morphological comparisons were carried out using standard taxonomic keys. DNA extracted from 12 Sri Lankan sandfly samples was PCR-amplified and sequenced for the variable region of Cytochrome oxidase subunit I. Existing DNA sequences of India from GenBank were utilized for a phylogenetic analysis between Sri Lanka and India. Salivary protein profiles were studied using SDS-PAGE, Western blot, and electrospray ionization/LC/MS/MS. The morphological similarities observed between female Ph. argentipes from India and Sri Lanka suggest the presence of Ph. argentipes var. glaucus. A phylogenetic analysis showed genetic divergence between Ph. argentipes populations, but both shared a similar salivary protein profile. A common, strong 30 kDa immunogenic band comprised PagSP05, PagSP06, and PagSP17 proteins of Ph. argentipes. The similarity between the immunogenic salivary proteins suggests their potential use as common markers for vector exposure or immune response stimulants across regions. The use of multiple samples for each category of serum would improve the comprehensiveness of the immunogenic profiles obtained.

Small dams drive Anopheles abundance during the dry season in a high malaria burden area of Malawi.

This study explores the influence of small dams on the exposure to malaria vectors during the dry season in Kasungu district, Malawi, an area recently identified as high priority for malaria interventions by the National Malaria Control Programme. Small dam impoundments provide communities with a continuous supply of water for domestic and agricultural activities across sub-Saharan Africa and are considered vital to food security and climate change resilience. However, these permanent water bodies also create ideal breeding sites for mosquitoes in typically arid landscapes. The study focuses on a specific dam impoundment and its vicinity, aiming to assess its spatial and temporal influence on indoor vector densities. From May to August 2021, CDC light traps were used to measure indoor mosquito densities for two consecutive nights per month in three communities located at increasing distances from the dam (0, ~1 and ~2 km). Simultaneously, drone imagery was captured for each community, enabling the identification of additional standing water within approximately 400 m of selected households. Larval sampling was carried out within the impoundment periphery and in additional water bodies identified in the drone imagery. Generalised linear mixed models (GLMMs) were employed to analyse the indoor Anopheles abundance data, estimating the effects of household structure (open/closed eaves), month, temperature and water proximity on malaria vector exposure. Throughout 685 trapping nights, a total of 1256 mosquitoes were captured, with 33% (412) being female Anopheles. Among these, 91% were morphologically identified as Anopheles funestus s.l., and 5% as Anopheles gambiae s.l. Catches progressively decline in each consecutive trapping month as the environment became drier. This decline was much slower in Malangano, the community next to the dam, with abundance being notably higher in June and July. Further, the majority of An. gambiae s.l. were caught in May, with none identified in July and August. Anopheles larvae were found both in the impoundment and other smaller water bodies such as irrigation wells in each survey month; however, the presence of these smaller water bodies did not have a significant impact on adult female mosquito catches in the GLMM. The study concludes that proximity to the dam impoundment was the primary driver of differences between survey communities with the abundance in Chikhombwe (~1 km away) and Chiponde (~2 km away) being 0.35 (95% confidence interval [CI], 0.19-0.66) and 0.28 (95% CI, 0.16-0.47) lower than Malangano, respectively, after adjusting for other factors. These findings underscore the importance of targeted interventions, such as larval source management or housing improvements, near small dams to mitigate malaria transmission risks during the dry season. Further research is needed to develop cost-effective strategies for vector control within and around these impoundments.

Update on Managing the Risks of Exposure to Lentiviral and Retroviral Vectors.

This paper aims to review the risks associated with using lentiviral and retroviral vectors in research and clinical settings and to propose an update to an effective treatment plan. Risks of exposure were evaluated based on vector design, safety features, viral tropism, transgene, and means and modes of transmission. These risks were weighed against the potential risks and benefits of current HIV medications. We recommend the following post-exposure prophylactic treatment for significant lentiviral vector exposures: 1) dolutegravir 50 mg. taken once a day for 7 days; and 2) tenofovir disoproxil fumarate 300 mg. taken once a day for 7 days (28 days of both medications for replication-competent vectors). Due to the highly efficient delivery of transgenes by modern lentiviral and retroviral vectors, post-exposure prophylaxis is indicated to prevent vector integration and oncogenic risks.

Seasonal dynamics of sand flies in Lençóis Maranhenses National Park: a tourist destination and endemic area for leishmaniasis.

Numerous sand fly species have been reported in Lençóis Maranhenses National Park (LMNP) in Northeast Brazil, including important Leishmania vectors, making the park an endemic area for tegumentary and visceral leishmaniasis. We evaluated sand fly abundance monthly over 7 years, correlating it with environmental variables and monthly tourist numbers in LMNP. Sand fly species were observed throughout the year, with Lutzomyia longipalpis Lutz & Neiva and Nyssomyia whitmani Antunes and Coutinho (Diptera: Psychodidae) being the most abundant species, especially from April to June. In addition to seasonal abundance patterns, Lu. longipalpis showed increasing abundance throughout 2013 until April 2014, whereas Ny. whitmani exhibited a consistent increase throughout the study period. Redundancy analysis indicated that monthly sand fly abundances increased with humidity but decreased with mean temperature and wind speed. Ecotourists mainly visit the park from May to September when interdune lagoons are full, coinciding with high-frequency vector activity during the rainy months (May-June). Tourists also visit in January and February during school holidays, when the rains begin and sand fly abundance increases, and in July, when sand fly abundance decreases. To date, no instances of infected tourists have been recorded, likely because visits to LMNP occur during the day when sand flies are inactive. However, there is a potential risk of vector exposure if tourists engage in nighttime cultural visits to villages around the park, where leishmaniasis cases occur annually.

Quasi-perfusion studies for intensified lentiviral vector production using a continuous stable producer cell line

Quasi-perfusion culture was employed to intensify lentiviral vector (LV) manufacturing using a continuous stable producer cell line in an 8-day process. Initial studies aimed to identify a scalable seeding density, with 3, 4, and 5 x 104 cells cm-2 providing similar specific productivities of infectious LV. Seeding at 3 x 104 cells cm-2 was selected, and the quasi-perfusion was modulated to minimise inhibitory metabolite accumulation and vector exposure at 37 °C. Similar specific productivities of infectious and physical LV were achieved at 1, 2, and 3 vessel volumes per day (VVD), with 1 VVD selected to minimise downstream processing volumes. The optimised process was scaled 50-fold to 1,264 cm2 flasks, achieving similar LV titres. However, scaling up beyond this to a 6,320 cm2 multilayer flask reduced titres, possibly from suboptimal gas exchange. Across three independent processes in 25 cm2 to 6,320 cm2 flasks, reproducibility was high with a coefficient of variation of (7.7 ± 2.9)% and (11.9 ± 3.0)% for infectious and physical LV titres, respectively. The optimised flask process was successfully transferred to the iCELLis™ Nano (Cytiva) fixed-bed bioreactor, with quasi-perfusion at 1 VVD yielding 1.62 x 108 TU.

Early induction of cytokine release syndrome by rapidly generated CAR T cells in preclinical models

Cytokine release syndrome (CRS) is a significant side-effect of conventional chimeric antigen receptor (CAR) T-cell therapy. To facilitate patient accessibility, short-term (st) CAR T cells, which are administered to patients only 24 h after vector exposure, are in focus of current investigations. Their impact on the incidence and severity of CRS has been poorly explored. Here, we evaluated CD19-specific stCAR T cells in preclinical models. In co-culture with tumor cells and monocytes, stCAR T cells exhibited anti-tumoral activity and potent release of CRS-related cytokines (IL-6, IFN-γ, TNF-α, GM-CSF, IL-2, IL-10). When administered to NSG-SGM3 mice, stCAR T cells, but not conventional CAR T cells, induced severe acute adverse events within 24 h, including hypothermia and weight loss, as well as high body scores, independent of the presence of tumor target cells. Human (IFN-γ, TNF-α, IL-2, IL-10) and murine (MCP-1, IL-6, G-CSF) cytokines, typical for severe CRS, were systemically elevated. Our data highlight potential safety risks of rapidly manufactured CAR T cells and suggest NSG-SGM3 mice as sensitive model for their preclinical safety evaluation.

Protective and curative effect of an experimental biocomplex against Flavescence dorée phytoplasma infection and transmission by Scaphoideus titanus under laboratory conditions

Flavescence dorée (FD) is one of the most devastating diseases threatening grapevine cultivation in Europe. The causal agents are phytoplasmas, which reside in phloem vessels of infected plants and in the body of leafhopper vectors, the most important being Scaphoideus titanus. Current control mainly relies on vector management, which is often insufficient to prevent from disease outbreaks. Suitable methods to interfere with phytoplasmas in planta are constantly sought to support insecticide sprays. We tested a biocomplex containing trace elements and soluble sugars, with potential systemic spread and antimicrobial activity, with the aim to verify its capability to penetrate plant phloem and to interfere with phytoplasma survival in vector-inoculated plants, using two FD isolates (FD-C and FD-D). The penetration into the plant phloem, as well as its acquisition by phloem-feeding vectors, were tested using an iridium-marked biocomplex. Phytoplasma infection was detected by real Time PCR and compared in plants exposed to biocomplex treatment before inoculation (short-time and mid-time preventive application), and to treatment after inoculation (curative application). We also assessed if the biocomplex interferes with the vector competence of S. titanus, by conducting phytoplasma transmission trials with sequential inoculation steps, where untreated and biocomplex-treated grapevine plantlets were sequentially offered to infectious leafhoppers. Foliar treatment with the biocomplex supported the penetration of phloem tissues, and the product was successfully acquired by the experimental phytoplasma vector Euscelidius variegatus after feeding on the phloem of treated plants. When the biocomplex was applied on grapevines, it induced a significant reduction of phytoplasma infection regardless of FD isolate, both as a preventive and curative treatment, indicating an antibacterial activity in the plant. Moreover, sequential inoculation experiments showed a decrease of S. titanus inoculation rates after vector exposure to a biocomplex-treated plant, suggesting a reduction of transmission efficiency that was higher using the FD-C rather than FD-D isolate. Our results indicate the tested biocomplex as a promising tool for the creation of anti-phytoplasma formulations to support vector management in FD containment.

Molecular prevalence of Anaplasma spp. in cattle and assessment of associated risk factors in Northeast Thailand.

Anaplasma spp. are common rickettsia species described in ruminant hosts, including cattle. The clinical signs of anaplasmosis range from asymptomatic to mortality. However, there are insufficient studies on epidemiology surveys of this blood pathogen. This study aimed to estimate the prevalence and risk factors of anaplasmosis in beef and dairy cattle in Northeast, Thailand. A total of 187 blood samples of beef and dairy cattle were collected from five provinces in Northeast Thailand. Anaplasma spp. infections were screened by microscopic examination and polymerase chain reaction targeting specific genes (msp4 gene for Anaplasma marginale and 16S rRNA gene for Anaplasma platys and Anaplasma bovis). Moreover, the associated risk factors for the infections were evaluated. Overall, blood samples from cattle revealed that 17.6% (33/187) were positive for Anaplasma spp. by microscopic examination and 20.8% (39/187) were positive by DNA amplification. Of these 20.8%, 17.6% were A. marginale and 3.2% were A. platys. However, A. bovis infection was not detected. Infection with Anaplasma spp. and A. marginale showed a significant association with breed and gender (p < 0.05) while age and packed cell volume levels showed no significant statistical relationship between Anaplasma spp. infected and uninfected animals. This study indicated that anaplasmosis is distributed in beef and dairy cattle in Thailand; therefore, prevention and control strategies for these pathogens should be improved. This information will benefit veterinarians and cowherds by avoiding vector exposure and eliminating tick breeding sites.

Exclusion of Horizontal and Vertical Transmission as Major Sources of Trypanosoma Cruzi Infections in a Breeding Colony of Rhesus Macaques (Macaca Mulatta).

The vector-borne protozoal parasite Trypanosoma cruzi causes Chagas disease in humans and animals. This parasite is endemic to the southern United States where outdoor-housed NHP at biomedical facilities are at risk of infection. In addi- tion to the direct morbidity caused by T. cruzi, infected animals are of limited biomedical research use because infections can produce confounding pathophysiologic changes even in animals with no clinical disease. In part due to concerns for direct T. cruzi transmission between animals, infected NHP at some institutions have been culled, removed, or otherwise isolated from uninfected animal populations. However, data that document horizontal or vertical transmission in captive NHP in the United States are not available. To evaluate the potential for inter-animal transmission and to identify environmental factors that affect the distribution of new infections in NHPs, we conducted a retrospective epidemiologic study of a rhesus macaque ( Macaca mulatta ) breeding colony in south Texas. We used archived biologic samples and husbandry records to identify the time and location of macaque seroconversion. These data were used to perform a spatial analysis of how geographic location and animal associations affected the spread of disease and to infer the importance of horizontal or vertical routes of transmission. The majority of T. cruzi infections were spatially clustered, suggesting that environmental factors promoted vector exposure in various areas of the facility. Although we cannot not rule out horizontal transmission, our data suggest that horizontal transmission was not a critical route for spread for the disease. Vertical transmission was not a contributing factor in this colony. In conclusion, our findings suggest that local triatome vectors were the major source of T. cruzi infections in captive macaques in our colony. Therefore, limiting contact with vectors, rather than segregation of infected macaques, is a key strategy for disease prevention at institutions that house macaques outdoors in the southern United States.

PK/PD Modeling to Inform Clinical Development of an Adeno-associated Virus Gene Transfer Therapy for Duchenne Muscular Dystrophy (S48.005)

<h3>Objective:</h3> To evaluate the pharmacokinetic (PK)/pharmacodynamic (PD) relationship between tissue vector genome exposure, biological efficacy and functional outcome in an animal model of Duchenne muscular dystrophy (DMD) (DMD^mdx mice) following treatment with delandistrogene moxeparvovec (SRP-9001). <h3>Background:</h3> Delandistrogene moxeparvovec is an investigational gene transfer therapy developed to address the root cause of DMD through targeted skeletal and cardiac muscle expression of SRP-9001 dystrophin protein, which contains key functional domains of dystrophin. <h3>Design/Methods:</h3> We evaluated vector biodistribution, expression, and clinical dose selection of delandistrogene moxeparvovec using a novel application of a PK and PD modeling approach applied to data collected from DMD^mdx mice. We analyzed PK/PD relationships between dose, tissue vector genome exposure, SRP-9001 dystrophin protein expression (percent dystrophin positive fibers [PDPF] and western blot), and functional improvement (relative specific force from tibialis anterior and diaphragm). <h3>Results:</h3> Linear kinetics with a dose-proportional increase in tissue drug exposure were demonstrated across the nearly 10-fold dose range (4.43×10¹³–4.01×10¹⁴ vg/kg), and in all tissues. The relationship between tissue vector exposure and PD endpoints (PDPF, motor function outcome) showed a saturable response across a wide range of vector exposures. The vector exposure at 1.33×10¹⁴ vg/kg (the clinical dose) approached the maximum treatment response. Relative specific force and PDPF were significantly correlated (<i>P</i>=4.43×10⁻⁶). However, the relationship appeared to be nonlinear, with increased PDPF expression approaching the maximal functional improvement. Relative specific force and western blot were not significantly correlated. <h3>Conclusions:</h3> For the first time, biodistribution, biomarker and functional efficacy data were used to quantify and demonstrate PK/PD relationships for an adeno-associated virus (AAV)-based gene transfer therapy in a DMD animal model. The results continue to support the expected therapeutic benefit and clinical dose of delandistrogene moxeparvovec, an AAV-based gene transfer therapy. <b>Disclosure:</b> Mrs. East has received personal compensation for serving as an employee of Sarepta Therapeutics. Rachael Potter has received stock or an ownership interest from Sarepta Therapeutics. Dr. Snedeker has received personal compensation for serving as an employee of Sarepta Therapeutics. Dr. Snedeker has stock in Sarepta Therapeutics. Dr. Haile has nothing to disclose. Christopher Wier has received personal compensation for serving as an employee of Sarepta Therapeutics. Christopher Wier has stock in Sarepta Therapeutics. Ms. Rodino-Klapac has received personal compensation for serving as an employee of Sarepta Therapeutics, Inc.. Ms. Rodino-Klapac has received stock or an ownership interest from Sarepta Therapeutics. Ms. Rodino-Klapac has received intellectual property interests from a discovery or technology relating to health care.

Analysis of Vector Shedding Following Treatment with Delandistrogene Moxeparvovec (SRP-9001), an Investigational rAAVrh74-Based Gene Therapy for Duchenne Muscular Dystrophy (DMD) (P5-8.013)

<h3>Objective:</h3> To evaluate extent and magnitude of vector shedding following delandistrogene moxeparvovec (SRP-9001) administration. <h3>Background:</h3> Delandistrogene moxeparvovec is an investigational gene transfer therapy developed to address the root cause of DMD through targeted skeletal and cardiac muscle expression of SRP-9001 dystrophin protein, which contains key functional domains of dystrophin. The hypothetical risk of seroconversion in naive individuals through exposure to vector shed from treated patients is a point to consider for healthcare providers and the Duchenne community. <h3>Design/Methods:</h3> We evaluated extent of vector shedding and clearance following a single administration of delandistrogene moxeparvovec (1.33×10¹⁴ vg/kg body weight) in ENDEAVOR (NCT04626674; Study 103; N=20), a Phase 1 study in patients with DMD. Delandistrogene moxeparvovec vector exposure in saliva, urine, and feces was quantified by droplet digital polymerase chain reaction. In a nonclinical study, we tested naive mice to determine risk of AAVrh74 seroconversion following mucosal vector exposure, with doses based on exposure levels (vector genome copies) demonstrated in nonclinical and clinical studies. Mice were exposed to AAVrh74.CMV.eGFP via optic exposure and intramuscularly. Antibody levels were measured by AAVrh74 ELISA at baseline and 4 weeks post-delivery. <h3>Results:</h3> Following treatment in ENDEAVOR, the percentage decrease in amount of vector shed was &gt;99% in saliva (n=12), urine (n=18), and feces (n=11) by Week 4. In mice, topical optical delivery of the AAVrh74 vector at relevant concentrations did not produce seropositivity or detectable vector genomes throughout tissues at doses based on clinical vector shedding levels. <h3>Conclusions:</h3> Clinical findings show that peak vector shedding occurs in the first few days after delandistrogene moxeparvovec administration and exponentially declines to insignificant levels by Week 4. In mice, seroconversion was not observed by relevant exposure route at relevant concentrations. Results suggest that the risk of seroconversion following exposure to vector shed by individuals treated with AAVrh74-based gene therapy may be very low. <b>Disclosure:</b> Jyoti Malhotra has nothing to disclose. Dr. Smith has received personal compensation for serving as an employee of Sarepta Therapeutics, Inc. Dr. Smith has stock in Sarepta Therapeutics. Sarah Lewis has received stock or an ownership interest from Sarepta. Mrs. Zhang has nothing to disclose. Mr. Asher has received personal compensation for serving as an employee of Sarepta Therapeutics. Mr. Asher has stock in Sarepta Therapeutics. Shufang Wang has received personal compensation for serving as an employee of Sarepta Therapeutics INC. Shufang Wang has received stock or an ownership interest from Sarepta Therapeutics INC. Mrs. East has received personal compensation for serving as an employee of Sarepta Therapeutics. Rachael Potter has received stock or an ownership interest from Sarepta Therapeutics. Ms. Rodino-Klapac has received personal compensation for serving as an employee of Sarepta Therapeutics, Inc.. Ms. Rodino-Klapac has received stock or an ownership interest from Sarepta Therapeutics. Ms. Rodino-Klapac has received intellectual property interests from a discovery or technology relating to health care.

Entomological parameters and population structure at a microgeographic scale of the main Colombian malaria vectors Anopheles albimanus and Anopheles nuneztovari.

Population subdivision among several neotropical malaria vectors has been widely evaluated; however, few studies have analyzed population variation at a microgeographic scale, wherein local environmental variables may lead to population differentiation. The aim of the present study was to evaluate the genetic and geometric morphometric structure of Anopheles nuneztovari and Anopheles albimanus in endemic localities of northwestern Colombia. Genetic and phenetic structures were evaluated using microsatellites markers and wing geometric morphometrics, respectively. In addition, entomological indices of importance in transmission were calculated. Results showed that the main biting peaks of Anopheles nuneztovari were between 20:00 and 22:00, whereas Anopheles albimanus exhibited more variation in biting times among localities. Infection in An. nuneztovari by Plasmodium spp. (IR: 4.35%) and the annual entomological inoculation rate (30.31), indicated high vector exposure and local transmission risk. We did not detect Plasmodium-infected An. albimanus in this study. In general, low genetic and phenetic subdivision among the populations of both vectors was detected using a combination of phenotypic, genetic and environmental data. The results indicated high regional gene flow, although local environmental characteristics may be influencing the wing conformation differentiation and behavioral variation observed in An. albimanus. Furthermore, the population subdivision detected by microsatellite markers for both species by Bayesian genetic analysis provides a more accurate picture of the current genetic structure in comparison to previous studies. Finally, the biting behavior variation observed for both vectors among localities suggests the need for continuous malaria vector surveys covering the endemic region to implement the most effective integrated local control interventions.

Spatial, environmental, and individual associations with Anopheles albimanus salivary antigen IgG in Haitian children.

IgG serology can be utilized to estimate exposure to Anopheline malaria vectors and the Plasmodium species they transmit. A multiplex bead-based assay simultaneously detected IgG to Anopheles albimanus salivary gland extract (SGE) and four Plasmodium falciparum antigens (CSP, LSA-1, PfAMA1, and PfMSP1) in 11,541 children enrolled at 350 schools across Haiti in 2016. Logistic regression estimated odds of an above-median anti-SGE IgG response adjusting for individual- and environmental-level covariates. Spatial analysis detected statistically significant clusters of schools with students having high anti-SGE IgG levels, and spatial interpolation estimated anti-SGE IgG levels in unsampled locations. Boys had 11% (95% CI: 0.81, 0.98) lower odds of high anti-SGE IgG compared to girls, and children seropositive for PfMSP1 had 53% (95% CI: 1.17, 2.00) higher odds compared to PfMSP1 seronegatives. Compared to the lowest elevation, quartiles 2-4 of higher elevation were associated with successively lower odds (0.81, 0.43, and 0.34, respectively) of high anti-SGE IgG. Seven significant clusters of schools were detected in Haiti, while spatially interpolated results provided a comprehensive picture of anti-SGE IgG levels in the study area. Exposure to malaria vectors by IgG serology with SGE is a proxy to approximate vector biting in children and identify risk factors for vector exposure.

VP.82 PK/PD modelling to inform clinical development of an adeno-associated virus gene transfer therapy for Duchenne muscular dystrophy

Delandistrogene moxeparvovec (SRP-9001) is an investigational gene transfer therapy for Duchenne muscular dystrophy (DMD) developed for targeted skeletal and cardiac muscle expression of micro-dystrophin—a shortened, functional dystrophin protein. We evaluated vector biodistribution, expression, and clinical dose selection of delandistrogene moxeparvovec using a novel application of a pharmacokinetic (PK) and pharmacodynamic (PD) modelling approach applied to data collected from an animal model of DMD (DMD^MDX mice). We analysed PK/PD relationships between dose, tissue vector genome exposure, micro-dystrophin protein expression (percentage dystrophin positive fibres [PDPF] and western blot), and functional improvement (relative specific force from tibialis anterior and diaphragm). Linear kinetics with a dose-proportional increase in tissue drug exposure was demonstrated across the nearly 10-fold dose range (4.43x10¹³–4.01x10¹⁴ vg/kg), and in all tissues. The relationship between tissue vector exposure and PD endpoints (PDPF, motor function outcome) showed a saturable response across a wide range of vector exposures. The vector exposure at 1.33x10¹⁴ vg/kg (the clinical dose) approached the maximum treatment response. Relative specific force and PDPF were significantly correlated (P=4.43x10^-6). However, the relationship appeared to be nonlinear, with increased PDPF expression approaching the maximal functional improvement. Relative specific force and western blot were not significantly correlated. For the first time, biodistribution, biomarker, and functional efficacy data were used to quantify and demonstrate PK/PD relationships for an adeno-associated virus (AAV)-based gene transfer therapy in a DMD animal model. The results continue to support the expected therapeutic benefit and clinical dose of delandistrogene moxeparvovec, an AAV-based gene transfer therapy.

High Prevalence of Plasmodium Infection in Fighting Cocks in Thailand Determined with a Molecular Method.

Avian malaria caused by Plasmodium and the malaria-like parasites of the genus Haemoproteus has been regularly described in multiple regions worldwide. These parasites significantly affect many avian taxa, including domestic chickens and fighting cocks. There are limited epidemiological studies of these blood parasites in vertebrate hosts, especially in Thailand. This study used microscopic examination of blood samples and PCR amplification exploiting primers for nucleotide sequences of Plasmodium or Haemoproteus species based on the cytochrome b gene to determine the occurrence of Plasmodium spp. in fighting cocks. Examination of 249 blood samples of fighting cocks revealed that 41.37% (103/249) were positive for malaria by microscopic examination and 88.76% (221/249) were positive by DNA amplification. Sequencing and DNA analysis of 61 PCR products revealed that infection by Plasmodium juxtanucleare was the most common avian malaria in fighting cocks in Thailand followed by infections by Plasmodium gallinaceum; however, Haemoproteus infection was not discovered. This study indicated that plasmodiasis is widespread in fighting cocks in Thailand although the prevalence was not clearly determined; therefore, prevention and control strategies for these protozoa should be improved, especially those for avoiding vector exposure and eliminating mosquito breeding sites.

Identification of potential inhibitors of Zika virus targeting NS3 helicase using molecular dynamics simulations and DFT studies.

Despite the various research efforts towards the drug discovery program for Zika virus treatment, no antiviral drugs or vaccines have yet been discovered. The spread of the mosquito vector and ZIKV infection exposure is expected to accelerate globally due to continuing global travel. The NS3-Hel is a non-structural protein part and involved in different functions such as polyprotein processing, genome replication, etc. It makes an NS3-Hel protein an attractive target for designing novel drugs for ZIKV treatment. This investigation identifies the novel, potent ZIKV inhibitors by virtual screening and elucidates the binding pattern using molecular docking and molecular dynamics simulation studies. The molecular dynamics simulation results indicate dynamic stability between protein and ligand complexes, and the structures keep significantly unchanged at the binding site during the simulation period. All inhibitors found within the acceptable range having drug-likeness properties. The synthetic feasibility score suggests that all screened inhibitors can be easily synthesizable. Therefore, possible inhibitors obtained from this study can be considered a potential inhibitor for NS3 Hel, and further, it could be provided as a lead for drug development.

Seroprevalence of vector-borne pathogens in outdoor workers from southern Italy and associated occupational risk factors

BackgroundVector-borne diseases (VBDs) represent an emerging global threat to public health due to the geographical expansion of arthropod vectors. The study aims to assess the seroprevalence of selected vector-borne pathogens (VBPs) in different groups of outdoor workers and the occupational risk factors for exposure to arthropod bites.MethodsA cross-sectional study was conducted on 170 workers recruited in two different regions of southern Italy, including farmers, forestry workers, veterinarians, geologists/agronomists and administrative employees, and tested for IgG antibodies against Bartonella henselae, Borrelia spp. Coxiella burnetii and Rickettsia conorii, using a chemiluminescent immunoassay (CLIA). The relationship among job characteristics, tick exposure and the prevalence of seropositive subjects for each pathogen was investigated by applying categorical principal component analysis (CATPCA).ResultsA high seroprevalence for C. burnetii (30.0%) and R. conorii (15.3%) was reported, mainly in farmers (67.7% and 54.8%, respectively) and forestry workers (29.0% and 16.1%, respectively), while a low prevalence was observed for B. henselae and Borrelia spp. (8.8% and 4.1%, respectively). The regression equation by CATPCA was significant for C. burnetii and R. conorii (P < 0.001), showing a positive association with job, tick bite exposure, working area and contact with animals.ConclusionsThese findings highlight the need of activating an appropriate occupational health response for minimizing the risk of arthropod vector exposure in workplaces, considering specific preventive measures in particular in high-risk job categories.Graphical

Mosquito alert: leveraging citizen science to create a GBIF mosquito occurrence dataset.

The MosquitoAlertdataset includes occurrence records of adult mosquitoes collected worldwide in 2014-2020 through Mosquito Alert, a citizen science system for investigating and managing disease-carrying mosquitoes. Records are linked to citizen science-submitted photographs and validated by entomologists to determine the presence of five targeted European mosquito vectors: Aedes albopictus, Ae. aegypti, Ae. japonicus, Ae. koreicus, and Culex pipiens. Most records are from Spain, reflecting Spanish national and regional funding, but since autumn 2020 substantial records from other European countries are included, thanks to volunteer entomologists coordinated by the AIM-COST Action, and to technological developments to increase scalability. Among other applications, the Mosquito Alert dataset will help develop citizen science-based early warning systems for mosquito-borne disease risk. It can also be reused for modelling vector exposure risk, or to train machine-learning detection and classification routines on the linked images, to assist with data validation and establishing automated alert systems.