Vector Transmission Research Articles

A reporter Oropouche virus expressing ZsGreen from the M segment enables pathogenesis studies in mice.

Oropouche fever caused by Oropouche virus (OROV) is a significant zoonosis in Central and South America. Despite its public health significance, we lack high-throughput diagnostics, therapeutics, and a comprehensive knowledge of OROV biology. Reporter viruses are valuable tools to rapidly study virus dynamics and develop neutralization and antiviral screening assays. OROV is a tri-segmented bunyavirus, which makes generating a reporter virus challenging, as introducing foreign elements into the viral genome typically affects fitness. We previously demonstrated that the non-structural gene NSm on the OROV medium (M) segment is non-essential for replication in vitro. Taking advantage of this, we have now generated a recombinant OROV expressing fluorescent protein ZsGreen in place of NSm. This reporter OROV is both stable and pathogenic in IFNAR-/- mice and provides a powerful tool for OROV pathogenesis studies and assay development.IMPORTANCEEmerging and reemerging infectious agents such as zoonotic bunyaviruses are of global health concern. Oropouche virus (OROV) causes recurring outbreaks of acute febrile illness in the Central and South American human populations. Biting midges are the primary transmission vectors, whereas sloths and non-human primates are their reservoir hosts. As global temperatures increase, we will likely see an expansion in arthropod-borne pathogens such as OROV. Therefore, developing reagents to study pathogen biology to aid in identifying druggable targets is essential. Here, we demonstrate the feasibility and use of a fluorescent OROV reporter in mice to study viral dynamics and pathogenesis. We show that this reporter OROV maintains characteristics such as growth and pathogenicity similar to the wild-type virus. Using this reporter virus, we can now develop methods to assist OROV studies and establish various high-throughput assays.

The Influential Influenza: The "Russian Catarrh" Pandemic of 1781-1782.

The influenza pandemic of 1781-1782 was remarkably well-documented, with investigations and treatment records produced in Britain, France, Germany, Italy, and Russia. Everyone agreed that outbreak began in St. Petersburg in December 1781 and then spread across northern Europe, but the medical communities' consensus did not solve all issues. Two questions would inspire years of debate. The first concerned the transmission vector of the pandemic: was it the result of neo-Hippocratic, miasmatic, or contagionist exposure? This was perhaps the greatest concern of the late-eighteenth century, and multiple physicians hoped the latest influenza pandemic could provide an answer. The second was no less difficult - where did the disease originate? This was not only because geography affected both prophylactic measures and treatment options but also produced diplomatic and commercial consequences. Was a quarantine necessary, preventing commercial exchanges? Did the risk of infection result in peace negotiations being delayed, potentially extending the American Revolution and the ongoing naval conflict in the Atlantic? Even if a consensus could be reached that this was a "Russian" catarrh, this would not resolve the method of disease transmission. The pandemic of 1781-1782 was not a turning point in the arguments among neo-Hippocratic, miasmatic, and contagionist physicians, but rather reveals all three positions could be held simultaneously.

Triatomine (Hemiptera: Reduviidae) populations and Trypanosoma cruzi genotyping in peridomestic and sylvatic environments in the semiarid region of Sergipe, Northeastern, Brazil

We assessed the diversity of triatomines, the rates of natural infection, and the discrete typing units (DTUs) of Trypanosoma cruzi isolated from them in two municipalities in the state of Sergipe, Brazil. Active searches for triatomines were conducted in the peridomicily and wild enviroments of 10 villages within the two municipalities. Triatomines were taxonomically identified and their feces were extracted using the abdominal compression method. Parasite detection was performed using optical microscopy. For Trypanosoma cruzi genotyping via PCR-FFLB, 151 samples of the subspecies Triatoma brasiliensis macromelasoma and Triatoma brasiliensis were isolated from both municipalities. In total, 505 triatomines were collected, with Triatoma brasiliensis macromelasoma being the most frequent species (58.81 %). Triatoma b. brasiliensis was the only species in both peridomestic and wild environments. Regarding the other species, T. pseudomaculata was found only in the peridomestic environment; and T. b. macromelasoma and Psammolestes tertius were found in the wild environment. Three Discrete Typing Units were identified: TcI (87.51 %) detected in T. b. brasiliensis and T. b. macromelasoma, TcI+TcIII (10.41 %) in T. b. macromelasoma, and TcI+Trypanosoma rangeli (2.08 %) in T. b. macromelasoma. It is concluded that T. b. macromelasoma is the species collected most frequently in the studied region and the one that presents the highest rates of natural infection, highlighting its epidemiological importance for the vectorial transmission of Chagas disease in Sergipe.

Demonstration of Insect Vector-Mediated Transfer of a Betasatellite between Two Helper Viruses.

Begomoviruses, transmitted by the whitefly Bemisia tabaci, pose significant threats to global agriculture due to their severe impact on various crops. Among the satellite molecules associated with begomoviruses, betasatellites play a crucial role in enhancing disease severity and yield losses. The spread and association of these molecules with helper viruses in host plants are thus matters of concern. Here, we focus on the propagation of betasatellites and, more specifically, on their transfer between different helper viruses and hosts through vector transmission. Our results show that the cotton leaf curl Gezira betasatellite (CLCuGeB), initially acquired with its helper virus cotton leaf curl Gezira virus (CLCuGeV) from an okra plant, can be transmitted and assisted by a different helper virus, tomato yellow leaf curl virus (TYLCV), in a different host plant (tomato plant). The new association can be formed whether TYLCV and CLCuGeB encounter each other in a host plant previously infected with TYLCV or in whiteflies having acquired the different components separately. Our findings reveal two pathways by which betasatellites can be transferred between helper viruses and host plants and highlight the ability of betasatellites to spread in begomovirus-infected environments.

Characterization of the microbiome of Aedes albopictus populations in different habitats from Spain and São Tomé

The mosquito microbiome significantly influences vector competence, including in Aedes albopictus, a globally invasive vector. Describing the microbiome and Wolbachia strains of Ae. albopictus from different regions can guide area-specific control strategies. Mosquito samples from Spain and São Tomé were analyzed using 16S rRNA gene sequencing and metagenomic sequencing. Wolbachia infection patterns were observed by sex and population. Female mosquitoes were blood-fed, a factor considered in analyzing their microbiota. Results revealed a dominance of dual Wolbachia infections, strains A and B, in the microbiome of both populations of Ae. albopictus, especially among females. Both populations shared a core microbiome, although 5 and 9 other genera were only present in Spain and São Tomé populations, respectively. Genera like Pelomonas and Nevskia were identified for the first time in Aedes mosquitoes. This study is the first to describe the Ae. albopictus bacteriome in Spain and São Tomé, offering insights for the development of targeted mosquito control strategies. Understanding the specific microbiome composition can help in designing more effective interventions, such as microbiome manipulation and Wolbachia-based approaches, to reduce vector competence and transmission potential of these mosquitoes.

An Overview of Parasitic Disease: Dracunculiasis

Dracunculiasis, also known as guinea worm disease, is a parasitic illness that only affects isolated, rural communities without access to clean drinking water in thirteen sub-Saharan African countries. The WHO has identified it as one of the next diseases to be eradicated. Water contaminated with copepods, or water fleas, carrying Dracunculiasis medinensis larvae is the vector of guinea worm disease transmission. A year after humans consume contaminated water, an adult female worm emerges, usually from a lower extremity, causing excruciating ulcers that can cause severe mobility impairment for several weeks. This illness strikes during the height of agricultural activity every year. A village's economically productive population is typically impacted in large numbers at the same time, which lowers agricultural output and causes economic difficulties. Since there is no known cure or vaccine for guinea worm disease, prevention is the key to its eradication. Globally, guinea worm disease cases have decreased by 98% since 1986, mostly due to community-based initiatives. Through these initiatives, the community has been taught how to filter water to get rid of parasites and how to stop people who have ulcers from contaminating drinking water sources. Sustained high-level political, financial, and community support will be necessary for complete eradication. By using an ayurvedic treatment approach, dracunculiasis can also be prevented. Ayurvedic medicine uses a few medicinal plants that exhibit antiparasitic activity against dracunculiasis, including Neem, Turmeric, Garlic, Amala, Baheda, Harde, Vidanga, Gokshura, Rasna, Bilva root, and Bala root.

First Report of a Phytoplasma Strain in the Elm Yellows Group (16SrV) associated with Virginia Creeper in Maryland, USA.

Virginia creeper (Parthenocissus quinquefolia [L.] Planch.) is a deciduous flowering vine in the Vitaceae family. Native to eastern North America, it is often used ornamentally as a climbing vine or as ground cover due to its rapid growth and foliage color in the fall. In July of 2022, along exterior walls of a private property in Lanham, MD, two Virginia creeper (VC) vines were observed displaying symptoms of yellow mottling and premature reddening of leaves. To investigate the cause of these symptoms, two symptomatic leaf samples and one asymptomatic leaf samples from a third vine in the same vicinity were collected for further analysis. A Qiagen DNeasy Plant Mini Kit was used to extract total DNA from leaf tissues according to the manufacturer's instructions. A phytoplasma-specific real-time PCR (Hodgetts et al. 2009) was used to test the DNA extracts, which detected the presence of phytoplasmas in the two DNA samples derived from symptomatic vines. The near full-length of the 16S ribosomal RNA gene was then amplified by seminested PCR from these samples with primers P1/16S-SR followed by P1A/16S-SR (Deng, and Hiruki 1991; Lee et al. 2004) and Sanger sequenced using primers P1A and 16S-SR. Analysis of the obtained 16S rDNA sequences revealed no variation between the two plant samples, and one sequence was deposited in GenBank representing the phytoplasma strain named VC-MD1 (GenBank PP746981). A BLASTn search of the 16S rRNA gene sequence in the NCBI nucleotide database, showed 99.93% sequence identity with the phytoplasma strain AldY-WA1 (GenBank MZ557341) from red alder in Washington, a phytoplasma associated with VC plants in southern Florida (GenBank AF305198) (Harrison et al. 2001), and other strains detected in grapevines in Europe described as "flavescence dorée" phytoplasma (GenBank AF176319) (Davis, and Dally 2001). The virtual restriction fragment length polymorphism pattern derived from iPhyClassifier (Zhao et al. 2009), indicated that VC-MD1 is indeed a member of the 16SrV-C phytoplasma subgroup. To confirm the identification, the partial spc operon and the partial tuf gene were amplified as previously described (Lee et al. 2010; Makarova et al. 2012). Specifically, the spc operon region was amplified using a nested PCR approach with primer set L15F1A-a/MapR1 followed by L15F1A-b/MapR1A-b. Sequence data obtained from the two loci were deposited to GenBank with accession numbers PP746982 (spc) and PP746983 (tuf). BLAST searches querying the nucleotide sequences of the spc operon and tuf gene showed 95.39% and 99.05% identity, respectively, to the corresponding loci of 'Candidatus Phytoplasma rubi' strain RS (GenBank CP114006) and hemp dogbane yellows phytoplasma strain HD1 (GenBank FR686506). Phylogenetic analysis based on secY and tuf gene sequences suggest that the VC-MD1 strain is evolutionary closest to 16SrV-C phytoplasma strains detected in various hosts in the United States, including HD1 and AldY-WA1. These North American strains cluster together on a distinct branch within the elm yellows group phytoplasmas. For the State of Maryland, this detection represents the first report of a phytoplasma strain member of the16SrV-C subgroup infecting VC plants. A phytoplasma of the same subgroup was previously detected in Florida in asymptomatic VC vines (Harrison et al. 2001). The 16S rRNA gene sequences of the two VC phytoplasma strains are nearly identical, differing by just a single nucleotide. The disease transmission vectors of the VC-MD1 strain and the prevalence of the disease in the region remains undetermined.

VL-HIV co-infection with Leishmania containing skin lesions resembling para-kala-azar dermal leishmaniasis.

Leishmaniases are a group of neglected vector-borne infectious diseases that are among the six priority endemic diseases worldwide. Visceral leishmaniasis (VL) is the most severe clinical manifestation, characterized by systemic and chronic visceral involvement and high mortality in immunosuppressed and untreated patients. VL can be complicated into post-kala-azar dermal leishmaniasis (PKDL), and when dermatologic disorders occur simultaneously with active VL, an intermediate clinical form called para-kala-azar dermal leishmaniasis (para-KDL) occurs. This clinical form is of great epidemiological relevance, as humans act as a source of infection for vectorial transmission. In the Americas, Brazil is among the seven countries responsible for more than 90% of VL cases, though reports of PKDL and para-KDL are rare. This paper presents three cases of VL-HIV co-infection with Leishmania-containing skin lesions resembling para-kala-azar dermal leishmaniasis. The cases were investigated by the team from the Infectious Diseases Department of University Hospital (HUMAP/UFMS) in Mato Grosso do Sul, Brazil. The three patients exhibited skin lesions where amastigote forms of L. (L.) infantum were identified. All cases exhibited similar clinical manifestations of para-KDL, including fever, hepatosplenomegaly, pancytopenia, and disseminated skin lesions. The study described the prevalence of comorbidities, the incidence of VL relapse, and the therapeutic regimen in relation to the outcomes. The study underscores the importance of follow-up and secondary prophylaxis in patients with VL, which are essential for the efficacy of the treatment. Furthermore, the study provides insight into the potential epidemiological profile of para-KDL cases in Brazil, which contributes to the development of more efficient clinical management strategies for patients.

16S rRNA metabarcoding for the identification of tick-borne bacteria in ticks in the Republic of Korea

Ticks are blood-sucking ectoparasites that act as vectors for transmission of various pathogens. The purpose of this study was to assess tick-borne bacteria, whether pathogenic or not, in ticks distributed in Korea using 16S rRNA metabarcoding and to confirm the results by PCR. Questing ticks were collected from four provinces in Korea in 2021 using the flagging method. After pooling the DNAs from the 61 tick pools (including 372 ticks), the bacterial 16S rRNA V3–V4 hypervariable region was amplified and sequenced using the MiSeq platform. Rickettsia, Ehrlichia, and the endosymbiont Wolbachia were confirmed by conventional PCR and molecular analysis. In total, 6907 ticks (534 pools) were collected and identified as belonging to five species (Haemaphysalis spp., H. longicornis, H. flava, I. nipponensis, and A. testudinarium). Through 16S rRNA metabarcoding, 240 amplicon sequence variants were identified. The dominant taxa were Rickettsiella and Coxiella. Additionally, pathogenic bacteria such as Rickettsia and Ehrlichia, endosymbiotic bacteria such as Wolbachia and Spiroplasma were identified. Polymerase chain reaction (PCR) was performed to confirm the presence of Rickettsia, Ehrlichia, Bartonella, and Wolbachia in individual ticks. Overall, 352 (65.92%) of 534 pools tested positive for at least one of the screened tick-borne bacteria. Rickettsia was the most prevalent (61.42%), followed by Wolbachia (5.05%). Ehrlichia was detected in 4.86% of tested samples, whereas Bartonella was not detected. In this study, 16S rRNA metabarcoding revealed the presence of Rickettsia, Wolbachia, and Ehrlichia, in that order of abundance, while showing absence of Bartonella. These results were confirmed to exhibit the same trend as that of the conventional PCR. Therefore, large-scale screening studies based on pooling, as applied in this study, will be useful for examining novel or rare pathogens present in various hosts and vectors.

Impacts of coingesting two tomato-infecting begomoviruses on virus infection in Bemisia tabaci and vector transmission

Impacts of coingesting two tomato-infecting begomoviruses on virus infection in Bemisia tabaci and vector transmission

Preparation and antibacterial activity of Ag-doped TiO2 coating on the porous surface of aluminum

Aluminum (Al) and its alloys have been widely used in daily life, but the absence of antibacterial properties makes them potential vectors for bacterial transmission. In this study, an Ag-doped TiO2 coating was successfully prepared on the porous surface of etched Al foil using the sol-gel method. The results indicated that the porous layer allowed the gel to permeate through the holes, thereby forming an interface carrying these antibacterial agents with a thickness of about 10 μm. The Ag dopant promoted the transformation of titania from rutile to anatase during the calcination process, with the rutile being absent when the atomic ratio of Ag to Ti reached 0.10. Meanwhile, the doped Ag+ was thermally decomposed into metallic Ag0. As a result, doping Ag enhanced the photocatalytic efficacy of TiO2 coating. Moreover, the antibacterial rate of this coating against Escherichia coli and Staphylococcus aureus under dark conditions could reach more than 99%, and the corrosion resistance of the Al foil was also slightly improved by it.

Effect of indoor residual spraying on sandfly abundance and incidence of visceral leishmaniasis in India, 2016–22: an interrupted time-series analysis and modelling study

Efforts to eliminate visceral leishmaniasis in India mainly consist of early detection and treatment of cases and indoor residual spraying with insecticides to kill the phlebotomine sandfly Phlebotomus argentipes that transmits the causative Leishmania protozoa. In this modelling study, we aimed to estimate the effect of indoor residual spraying (IRS) on vector abundance and transmission of visceral leishmaniasis in India. In this time-series analysis and modelling study, we assessed the effect of IRS on vector abundance by using indoor vector-abundance data (from 2016 to 2022) and IRS quality-assurance data (from 2017-20) from 50 villages in eight endemic blocks in India where IRS was implemented programmatically. To assess a potential dose-response relation between insecticide concentrations and changes in sandfly abundance, we examined the correlation between site-level insecticide concentrations and the site-level data for monthly sandfly abundances. We used mathematical modelling to link vector data to visceral leishmaniasis case numbers from the national Kala-Azar Management Information System registry (2013-21), and to predict the effect of IRS on numbers of averted cases and deaths. IRS was estimated to reduce indoor sandfly abundance by 27% (95% CI 20-34). Concentrations of insecticides on walls were significantly-but weakly-associated with the degree of reduction in vector abundance, with a reduction of -0·0023 (95% CI -0·0040 to -0·0007) sandflies per mg/m2 insecticide (p=0·0057). Reported case numbers of visceral leishmaniasis were well explained by trends in vector abundance. Village-wide IRS in response to a newly detected case of visceral leishmaniasis was predicted to reduce disease incidence by 6-40% depending on the presumed reduction in vector abundance modelled. Indoor residual spraying has substantially reduced sandfly abundance in India, which has contributed to reductions in visceral leishmaniasis and related deaths. To prevent the re-emergence of visceral leishmaniasis as a public health problem, surveillance of transmission and sandfly abundance is warranted. Bill & Melinda Gates Foundation. For the Hindi translation of the abstract see Supplementary Materials section.

Abundance and Parasitic Load of Housefly (Musca Domestica) in Amansea Area of Anambra State, Nigeria

Houseflies (Musca domestica) are major transmitters of pathogenic organisms to humans worldwide. This study was carried out to determine the relative abundance of housefly and its parasitic load in four different sites in the Amansea area of Anambra state, Nigeria. The housefly samples were collected with the aid of fly trap and sweep net and analyzed using sedimentation methods. Data obtained was calculated and expressed as simple percentages while chi-square was used to test for differences in prevalence rates of parasites in the study areas. Out of the 259 houseflies examined, 74 were infested with a prevalence of 28.57%. Houseflies were more abundant in the abattoir 92(35.52) followed by market area 71(27.41%), refuse dump sites (19.69%) while holy family hostels recorded the lowest abundance (17.37) (P=0.000). Houseflies infested with parasites were mostly found in holy family hostel 31(68.89) while the market area had the least infested house flies 5(7.04%). Two protozoan parasites (Entamoeba histolytica and Entamoeba coli) and two nematodes (Strongyloides stercoralis and Ascaris lumbricoides) were the parasites seen from the microscopic examination of samples. E. histolytica (7.61%) and E. coli (5.43%) were the parasites seen in Abattoir while S. stercoralis (7.04%) was the only parasite in the market area. E. histolytica (17.78%) and A. lumbricoides (51.11%) were seen in holy family hostel while E. coli (11.76%), S. stercoralis (7.84%) and A. lumbricoides (31.37%) were the parasites recovered from the houseflies in the Refuse dump site. The infestation of houseflies in the area is a threat to human health due to their ability to act as mechanical vectors for transmission of infectious pathogens. Hence, proper disposal and prompt removal of waste by environmental officers should be prioritized in the area as well as mass education of the public on the health implications of poor hygiene and poor sanitary practices.

GIS-ODE: linking dynamic population models with GIS to predict pathogen vector abundance across a country under climate change scenarios.

Mechanistic mathematical models such as ordinary differential equations (ODEs) have a long history for their use in describing population dynamics and determining estimates of key parameters that summarize the potential growth or decline of a population over time. More recently, geographic information systems (GIS) have become important tools to provide a visual representation of statistically determined parameters and environmental features over space. Here, we combine these tools to form a 'GIS-ODE' approach to generate spatiotemporal maps predicting how projected changes in thermal climate may affect population densities and, uniquely, population dynamics of Ixodes ricinus, an important tick vector of several human pathogens. Assuming habitat and host densities are not greatly affected by climate warming, the GIS-ODE model predicted that, even under the lowest projected temperature increase, I. ricinus nymph densities could increase by 26-99% in Scotland, depending on the habitat and climate of the location. Our GIS-ODE model provides the vector-borne disease research community with a framework option to produce predictive, spatially explicit risk maps based on a mechanistic understanding of vector and vector-borne disease transmission dynamics.

The evolutionary and molecular history of a chikungunya virus outbreak lineage.

In 2018-2019, Thailand experienced a nationwide spread of chikungunya virus (CHIKV), with approximately 15,000 confirmed cases of disease reported. Here, we investigated the evolutionary and molecular history of the East/Central/South African (ECSA) genotype to determine the origins of the 2018-2019 CHIKV outbreak in Thailand. This was done using newly sequenced clinical samples from travellers returning to Sweden from Thailand in late 2018 and early 2019 and previously published genome sequences. Our phylogeographic analysis showed that before the outbreak in Thailand, the Indian Ocean lineage (IOL) found within the ESCA, had evolved and circulated in East Africa, South Asia, and Southeast Asia for about 15 years. In the first half of 2017, an introduction occurred into Thailand from another South Asian country, most likely Bangladesh, which subsequently developed into a large outbreak in Thailand with export to neighbouring countries. Based on comparative phylogenetic analyses of the complete CHIKV genome and protein modelling, we identified several mutations in the E1/E2 spike complex, such as E1 K211E and E2 V264A, which are highly relevant as they may lead to changes in vector competence, transmission efficiency and pathogenicity of the virus. A number of mutations (E2 G205S, Nsp3 D372E, Nsp2 V793A), that emerged shortly before the outbreak of the virus in Thailand in 2018 may have altered antibody binding and recognition due to their position. This study not only improves our understanding of the factors contributing to the epidemic in Southeast Asia, but also has implications for the development of effective response strategies and the potential development of new vaccines.

Antimicrobial activity of NK cells to Trypanosoma cruzi infected human primary Keratinocytes.

Infection with the protozoan parasite Trypanosoma cruzi is causative for Chagas disease, which is a highly neglected tropical disease prevalent in Latin America. Humans are primary infected through vectorial transmission by blood-sucking triatomine bugs. The parasite enters the human host through mucous membranes or small skin lesions. Since keratinocytes are the predominant cell type in the epidermis, they play a critical role in detecting disruptions in homeostasis and aiding in pathogen elimination by the immune system in the human skin as alternative antigen-presenting cells. Interestingly, keratinocytes also act as a reservoir for T. cruzi, as the skin has been identified as a major site of persistent infection in mice with chronic Chagas disease. Moreover, there are reports of the emergence of T. cruzi amastigote nests in the skin of immunocompromised individuals who are experiencing reactivation of Chagas disease. This observation implies that the skin may serve as a site for persistent parasite presence during chronic human infection too and underscores the significance of investigating the interactions between T. cruzi and skin cells. Consequently, the primary objective of this study was to establish and characterize the infection kinetics in human primary epidermal keratinocytes (hPEK). Our investigation focused on surface molecules that either facilitated or hindered the activation of natural killer (NK) cells, which play a crucial role in controlling the infection. To simulate the in vivo situation in humans, an autologous co-culture model was developed to examine the interactions between T. cruzi infected keratinocytes and NK cells. We evaluated the degranulation, cytokine production, and cytotoxicity of NK cells in response to the infected keratinocytes. We observed a strong activation of NK cells by infected keratinocytes, despite minimal alterations in the expression of activating or inhibitory ligands on NK cell receptors. However, stimulation with recombinant interferon-gamma (IFN-γ), a cytokine known to be present in significant quantities during chronic T. cruzi infections in the host, resulted in a substantial upregulation of these ligands on primary keratinocytes. Overall, our findings suggest the crucial role of NK cells in controlling acute T. cruzi infection in the upper layer of the skin and shed light on keratinocytes as potential initial targets of infection.

Blood biodistribution and vector shedding of valoctocogene roxaparvovec in people with severe hemophilia A

AbstractFollowing systemically administered adeno-associated virus gene therapy, vector particles are widely distributed, raising concerns about horizontal or germline vector transmission. Characterization of biodistribution and kinetics of vector DNA in body fluids can address these concerns and provide insights into vector behavior in accessible samples. We investigated biodistribution and vector shedding profile of valoctocogene roxaparvovec in men with severe hemophilia A enrolled in the phase 3 GENEr8-1 trial. Participants (n = 134) received a single 6 × 1013 vector genome (vg)/kg infusion and were assessed over 3 years. Vector DNA was measured using 4 different assays. Total vector DNA was evaluated in blood, saliva, stool, semen, and urine by quantitative polymerase chain reaction (qPCR). Encapsidated vector DNA was measured in plasma and semen with immunocapture-based qPCR. Contiguity of vgs and assembly of inverted terminal repeat fusions were measured in whole blood and peripheral blood mononuclear cells (PBMCs) using multicolor digital PCR. Median peak vector DNA levels observed 1 to 8 days after dosing were highest in blood, followed by saliva, semen, stool, and urine. Concentrations declined steadily. Encapsidated vector DNA cleared faster than total vector DNA, achieving clearance by ≤12 weeks in plasma and semen. Predominant vector genome forms transitioned from noncontiguous to full-length over time in whole blood and PBMCs, indicating formation of stable circularized episomes within nucleated cells. The replication-incompetent nature of valoctocogene roxaparvovec, coupled with steady clearance of total and encapsidated vector DNA from shedding matrices, indicates transmission risk is low. This trial was registered at www.ClinicalTrials.gov as #NCT03370913.

Exosomal miR-7-25207 Increases Subgroup J Avian Leukosis Virus Titers by Targeting the Akt-CyclinQ1 and PRC1-YAF2 Dual Pathways.

Subgroup J avian leukosis virus (ALV-J) is a major pathogen in poultry, causing substantial economic losses to the poultry industry worldwide. Exosomal small RNAs derived from virus-infected cells or biological fluids can serve as viral transmission vectors. However, the role and mechanism of exosomal miRNA in ALV-J infection are unclear. In this study, we demonstrated that exosomal microRNA-7-25207 (miR-7-25207) could increase the titers of ALV-J. Exosomes isolated from ALV-J-infected DF-1 cells (Exo-ALV-J) contained partial viral proteins from ALV-J and could transmit the infection to uninfected DF-1 cells, leading to productive infection. Additionally, the RNA expression profile of exosomes was altered following ALV-J infection. miRNA analysis revealed that the expression of exosomal miR-7-25207 increased. Overexpression of miR-7-25207 significantly increased the titers of ALV-J in transfected cells. Furthermore, miR-7-25207 directly suppressed the expression of Akt and PRC1. Akt, in turn, directly inhibited CyclinQ1 expression, while PRC1 directly interfered with YAF2 expression. In conclusion, ALV-J infection activates the expression of miR-7-25207, which is subsequently delivered via exosomes to uninfected cells, increasing ALV-J titers by targeting Akt-CyclinQ1 and PRC1-YAF2 dual pathways. These findings suggest that exosomal miR-7-25207 may serve as a potential biomarker for clinical parameters in ALV-J infection.

Gambaran Kepadatan Dan Identifikasi Lalat (Diptera) Di Pasar Waiheru Perumnas Kecamatan Baguala Kota Ambon

Flies are vectors for disease transmission, especially mechanically and through their vomit and feces. Items, especially food, are usually contaminated by human feces, rubbish, the saliva of sick people, scars from animal carcasses and other things that are covered in flies. Flies can throw feces on food so that the food becomes contaminated with fly eggs and larvae. Disruption of comfort, eyesore, itching of the skin, causing discomfort, decreased appetite. To determine the density and identification of flies in the Waiheru Perumnas market, Baguala District, Ambon City. Type of descriptive research that describes a condition based on observation and examination of the Ambon Environmental Health Engineering Center (BTKL) Laboratory. The aim of descriptive research is to describe the density and identification of flies in the Waiheru Perumnas market, Baguala District, Ambon City. Measuring the density of flies in the Waiheru Perumnas market, Baguala District, Ambon City is based on 3 points, 10 measurements at each point in the morning, at the first point (los fish) amounted to 87 flies, at point II (vegetable stall there were 55 flies and at point III (fruit stall) there were 45 flies. Identification of fly types resulted in 3 types of flies identified, namely Musca Domestica (house fly), Cliphora Sp (green flies) and Sarcophaga (meat flies). The density of flies in the Waiheru Perumnas market, Baguala District, Ambon City, based on the installation of fly grills based on the 10 highest points, was 187 flies and at the 5 highest points was 134 flies with 23 flies caught in the fly trap. From the results obtained, it can be seen that the density of flies is very high so it is necessary to deal with fly breeding sites and fly control measures.

Prevalence analysis of Chagas disease by age group in an endemic region of Brazil: possible scenario of active vectorial transmission

ObjectivesChagas disease (CD) is an infectious disease that predominantly affects poor and vulnerable populations. The last estimate conducted by the World Health Organization in Latin America regarding the prevalence of CD occurred more than 10 years ago. However, there is a scarcity of data assessing the magnitude of CD in populations residing in considered high-risk regions. Therefore, this study aimed to assess the seroprevalence of CD in an endemic region in Northern Minas Gerais through serologic screening. MethodsThis is a prevalence study conducted in the municipalities of Catuti, Mato Verde, Mirabela, Montes Azul, and São Francisco, Minas Gerais, Brazil. Data collection occurred between December 2021 and December 2022, involving a questionnaire with closed-ended questions. The variables analyzed included serologic test results, stratified age groups, health indicators, and housing conditions. ResultsOf the 2978 participants, 272 individuals (9.1%) tested positive for CD serology. In the age group of 4 to 14 years, 15 to 49 years, and 50 years or older, the prevalence of positive serology was 0.8% (95% confidence interval [CI] 0.16-1.43), 5.5% (95% CI 4.20-6.83), and 18.8% (95% CI 16.48-21.11), respectively. Among the participating municipalities, Mato Verde had the highest prevalence of positive serology for CD (17%). For participants aged 4 to 14 years with positive serology for CD, first-degree relatives were invited to undergo serologic testing. It was possible to collect samples from relatives of all participants in this age group. However, none of the relatives tested positive. ConclusionThis study identified a 9.1% prevalence of individuals affected by CD who were unaware of their condition. In addition, having infected children in the 4 to 14 age group with mothers with negative serology would rule out congenital transmission of the disease.