Vector Host Research Articles

Mapping the risk of exposure to Crimean-Congo haemorrhagic fever virus in the Iberian Peninsula using Eurasian wild boar (Sus scrofa) as a model

Crimean-Congo haemorrhagic fever (CCHF) virus (CCHFV) is a tick-borne zoonotic pathogen that can cause a lethal haemorrhagic disease in humans. Although the virus appears to be endemically established in the Iberian Peninsula, CCHF is an emerging disease in Spain. Clinical signs of CCHFV infection are mainly manifested in humans, but the virus replicates in several animal species. Understanding the determinants of CCHFV exposure risk from animal models is essential to predicting high-risk exposure hotspots for public health action. With this objective in mind, we designed a cross-sectional study of Eurasian wild boar (Sus scrofa) in Spain and Portugal. The study analysed 5,291 sera collected between 2006 and 2022 from 90 wild boar populations with a specific double-antigen ELISA to estimate CCHFV serum prevalence and identify the main determinants of exposure probability. To do so, we statistically modelled exposure risk with host- and environment-related predictors and spatially projected it at a 10 × 10 km square resolution at the scale of the Iberian Peninsula to map foci of infection risk. Fifty-seven (63.3 %) of the 90 populations had at least one seropositive animal, with seroprevalence ranging from 0.0 to 88.2 %. Anti-CCHFV antibodies were found in 1,026 of 5,291 wild boar (19.4 %; 95 % confidence interval: 18.3–20.5 %), with highest exposure rates in southwestern Iberia. The most relevant predictors of virus exposure risk were wild boar abundance, local rainfall regime, shrub cover, winter air temperature and soil temperature variation. The spatial projection of the best-fit model identified high-risk foci as occurring in most of western and southwestern Iberia and identified recently confirmed risk foci in eastern Spain. The results of the study demonstrate that serological surveys of CCHFV vector hosts are a powerful, robust and highly informative tool for public health authorities to take action to prevent human cases of CCHF in enzootic and emergency settings.

Experimental evidence reveals that vector host preference and performance across host plants is not altered by vector-borne plant viruses

Many plant pathogens are transmitted by generalist insects that move between host species, and vector preference and performance on different hosts affects pathogen transmission. Most studies of host–vector–pathogen interactions focus on a single host, however, and it is relatively unknown if viruses affect the host breadth of vectors. For example, although pea aphids (Acyrthosiphon pisum) often form tight associations with specific hosts, viruses transmitted by aphids can break down host defenses, potentially weakening these associations and increasing the host breadth of their vectors. Here, we assessed vector host preference and performance in an experiment with five pea aphid colonies with distinct host associations, five host species, and two viruses (Pea enation mosaic virus [PEMV] and Bean leafroll virus [BLRV]). Our prediction was that pea aphids would have higher performance on their inferior hosts and be less discriminating among hosts when those are infected, compared with noninfected hosts. Our results show that aphid performance, measured as the rate of increase on hosts, differed among hosts based on their host association, and relative performance among hosts was indeed altered by virus infection. Aphid preferences, measured using a choice bioassay in which aphids could settle on any of the five host species, also differed among aphid colonies but was not affected by virus infection status. We also did not find that infection status of hosts altered the performance of aphids on their preferred host. Furthermore, virus infection did not disrupt preference performance relationships for the aphids. Overall, our study suggests that host preference and performance are deeply ingrained in pea aphids, and that host associations may be robust to the presence of host-manipulating plant pathogens.

Predicting the potential worldwide distribution of Aedes aegypti under climate change scenarios

Background: Climate change is one of the most important factors associated with medically important insect pests such as mosquitoes (Diptera: Culicidae). Diseases spread by mosquitoes are increasing due to changes in global temperature and weather patterns that are altering vector host ranges allowing spread into new regions. Zika, dengue fever, chikungunya and yellow fever are arboviral infections that are spread by Aedes aegypti (Culicidae). The objective of the current research is to study the potential geographic distribution habitats of Ae. aegypti in the world under current and future climate conditions. Methods: Data of Ae. aegypti was obtained from the global biodiversity information facility and used 19 bioclimatic layers (bio01-bio19) and elevation from the WorldClim database. The scenarios used are the Beijing climate center climate system model (BCC-CSM2-MR) and the institute Pierre-Simon Laplace, coupled model intercomparison project (IPSL-CM6A-LR) with two shared socio-economic pathways (SSPs) for each of the general circulation model (GCMs): SSP126 and SSP585. Results: The results revealed that altitude, temperature, seasonality (standard deviation *100) (bio4), and annual precipitation (bio12) were the most important environmental variables that affect the distribution of Ae. aegypti. Conclusions: The models showed that Africa and South America maintained very high and excellent habitat suitability for Ae. Aegypti under the current potential distribution map.

Conflict-aware workload co-execution on SX-aurora TSUBASA

NEC SX-Aurora TSUBASA (SX-AT) is the latest vector supercomputer, consisting of host processors called Vector Hosts (VHs) and vector processors called Vector Engines (VEs). The goal of this work is to simultaneously use both VHs and VEs to increase the resource utilization and improve the system throughput by co-executing more workloads. One difficulty is that performance interferences among VH and VE workloads could occur because they share some computing resources and potentially compete to use the same resource at the same time, so-called resource conflicts. To achieve efficient workload co-execution, first, this paper experimentally investigates the performance interference between a VH and a VE, when each of the two processors executes a different workload. It is empirically shown that the frequency of system calls from the VE workload could be a good indicator to predict if the co-execution could cause severe performance interference, even though monitoring system calls requires a huge runtime overhead and it is impractical to simply use it for decision making of co-execution. Then, this paper proposes a workload co-execution strategy based on a practical approach to identifying a pair of VE and VH workloads that could cause severe performance interferences. Our evaluation results clearly demonstrate that the system call frequency can be used to predict if the workload can affect the performance of another co-executing workload, and VH’s CPU load can be a good approximation of the system call frequency. The proposed approach based on the CPU loads could accurately identify a pair of workloads causing frequent resource conflicts, and thus reduce the risk of severe performance interferences between co-executing workloads on an SX-AT system, resulting in shorter makespan without significantly increasing the turn-around time.

Mitochondrial genome amplification of avian haemosporidian parasites from single-infected wildlife samples using a novel nested PCR approach

Haemosporidian parasites that infect birds (Apicomplexa: Haemosporida) are blood parasites that require an invertebrate host (vector) and a vertebrate host for their lifecycle and cause malaria-like diseases. This group of parasites has provided valuable insights into host specificity, virulence, and parasite dispersal. Additionally, they have played a significant role in reshaping our understanding of the evolutionary history of apicomplexans. In order to accurately identify species and to address phylogenetic questions such as the timing of the haemosporidian radiation, the use of a sufficiently large genetic data set is crucial. However, acquiring this genetic data poses significant challenges. In this research, a sensitive nested PCR assay was developed. This assay allows for the easy amplification of complete mitochondrial genomes of haemosporidian parasites in birds, even during the chronic stage of infection. The effectiveness of this new nested PCR assay was evaluated using blood and tissue samples of birds with verified single parasite infections from previous studies. The approach involves amplifying four overlapping fragments of the mitochondrial genome and requires DNA extracts from single-infected samples. This method successfully amplified the complete mitochondrial genomes of 24 distinct haemosporidian parasite lineages found in various bird species. This data is invaluable for conducting phylogenetic analyses and accurately defining species. Furthermore, this study proposes the existence of at least 15 new haemosporidian parasite species based on the genetic information obtained. Data regarding pGRW04, previously categorized as Plasmodium relictum like pSGS1 and pGRW11, indicates that the pGRW04 lineage is actually a separate, hidden Plasmodium species.

Tick-to-host transmission differs between Borrelia afzelii strains.

Many vector-borne pathogens establish multiple-strain infections in the vertebrate host and the arthropod vector. Multiple-strain infections in the host influence strain acquisition by naive vectors. Whether multiple-strain infections in the vector influence strain-specific transmission to naive hosts remains unknown. The spirochete, Borrelia afzelii, causes Lyme borreliosis and multiple-strain infections are common in both the tick vector and vertebrate host. Our study used two B. afzelii strains: Fin-Jyv-A3 and NE4049. Donor mice were infected with Fin-Jyv-A3 alone, NE4049 alone, or with both strains. Larval ticks fed on donor mice and molted into nymphal ticks infected with either strain or both strains. These nymphs were fed on test mice to determine whether multiple-strain infections in the nymph influence nymph-to-host transmission (NHT). Multiple-strain infection in the donor mice reduced the acquisition of both strains by ticks by 23%. Thus, a substantial fraction of infected nymphs from the multiple strain treatment were infected with the "wrong" competitor strain rather than the "right" focal strain. As a result, nymphs from the multiple strain treatment were 46% less likely to infect the test mice with the focal strain compared to nymphs from the single strain treatment. However, multiple-strain infection in the nymphal tick had no effect on the NHT of either strain. The nymphal spirochete load of Fin-Jyv-A3 was 1.9 times higher compared to NE4049. NHT of Fin-Jyv-A3 (79%) was 1.5 times higher compared to NE4049 (53%). Our study suggests that B. afzelii strains with higher nymphal spirochete loads have higher NHT. IMPORTANCE For many vector-borne pathogens, multiple-strain infections in the vertebrate host or arthropod vector are common. Multiple-strain infections in the host reduce strain acquisition by feeding vectors. Whether multiple-strain infections in the vector influence strain transmission to the host remains unknown. In our study, we used two strains of the tick-borne spirochete Borrelia afzelii, which causes Lyme borreliosis, to investigate whether multiple-strain infections in the nymphal tick influenced nymph-to-host transmission (NHT) of strains. Multiple-strain infections in mice reduced the acquisition of both B. afzelii strains by nymphal ticks. As a result, nymphs from the multiple strain treatment were less likely to infect naive test mice with the focal strain. Multiple-strain infection in the nymphal ticks did not influence the NHT of either strain. The strain with the higher bacterial abundance in the nymph had higher NHT. Our study suggests that pathogen abundance in the arthropod vector is important for vector-to-host transmission.

Simultaneous detection and quantification by multiplex qPCR of 'Candidatus Phytoplasma asteris' and 'Candidatus Phytoplasma fraxini' in a plant host and insect vectors

AbstractPhytoplasmas are bacteria transmitted by insects that can cause plant diseases. In Bogotá 'Candidatus Phytoplasma asteris' and ' Candidatus Phytoplasma fraxini', infect 11 species of urban trees, weeds, grass, potato and strawberry. A set of primers, that amplify both phytoplasmas species were designed and used for absolute and relative qPCR quantification of the 16SrRNA gene. The primers AJ-16Sr-F/AJ-16Sr-R allowed the amplification of ‘Ca. P. asteris’, ‘Candidatus Phytoplasma palmae’, ‘Ca. P. fraxini’ and ‘Candidatus Phytoplasma phoenicium’, not of ‘Candidatus Phytoplasma pruni’. Absolute qPCR detected phytoplasmas between 1 × 109 and 1 × 103 copies/μL DNA extract. Two species-specific hydrolysis probes, AJ-16SrI-Cy5.5 and AJ-16SrVII-TexRed, were designed to detect 'Ca. P. asteris' and 'Ca. P. fraxini' respectively, using the AJ-16Sr-F/AJ-16Sr-R primers. For relative quantification, the 18SrRNA gene was used as normalizer. Relative qPCR detected phytoplasmas between 1 × 109 and 1 × 103 copies/μL DNA extract. Multiplex reactions allowed the specific quantification of 'Ca. P. asteris', 'Ca. P. fraxini' in comparison to the normalizer. qPCR methods were validated on natural hosts Andean oak trees and leafhoppers. The relative quantification values were higher for 'Ca. P. fraxini' (x̅ RQ = 3203.1 ± 2622,9 n = 14) compared with 'Ca. P. asteris' (x̅ RQ = 14.9 ± 24,5 n = 6) in oak tree samples. In the leafhoppers, the relative quantification values ranged between RQ = 26.5 and RQ = 294,927.3 for 'Ca. P. fraxini’ and RQ = 34.8 and RQ = 1722.2 for 'Ca. P. asteris'. In conclusion, although absolute qPCR allowed the quantification of phytoplasmas by comparing Cq (quantification cycle) values of samples with a standard curve, it did not allow to differentiate between 'Ca. P. asteris' and 'Ca. P. fraxini'. In contrast, relative qPCR assays using specific hydrolysis probes allowed the specific detection and quantification of each phytoplasma, in individual and mixed infections in insect vectors and plant hosts.

Turnip yellows virus variants differ in host range, transmissibility, and virulence.

Turnip yellows virus (TuYV; family Solemoviridae, genus Polerovirus, species Turnip yellows virus) is a genetically diverse virus that infects a broad range of plant species across the world. Due to its global economic significance, most attention has been given to the impact of TuYV on canola (syn. oilseed rape; Brassica napus). In Australia, a major canola-exporting country, TuYV isolates are highly diverse, with the most variation concentrated in open reading frame 5 (ORF 5), which encodes the readthrough domain (P5) component of the readthrough protein (P3P5), which plays an important role in host adaptation and aphid transmission. When analysing ORF 5, Australian TuYV isolates form three phylogenetic groups with just 45 to 49% amino acid sequence identity: variants P5-I, P5-II, and P5-III. Despite the possible implications for TuYV epidemiology and management, research examining phenotypic differences between TuYV variants is scarce. This study was designed to test the hypothesis that three TuYV isolates, representing each of the Australian P5 variants, differ phenotypically. In particular, the host range, vector species, transmissibility, and virulence of isolates 5414 (P5-I5414), 5509 (P5-II5509), and 5594 (P5-III5594) were examined in a series of glasshouse experiments. Only P5-I5414 readily infected faba bean (Vicia faba), only P5-II5509 infected chickpea (Cicer arietinum), and only P5-I5414 and P5-III5594 infected lettuce (Lactuca sativa). Myzus persicae transmitted each isolate, but Brevicoryne brassicae and Lipaphis pseudobrassicae did not. When using individual M. persicae to inoculate canola seedlings, P5-I5414 had significantly higher transmission rates (82%) than P5-II5509 (62%) and P5-III5594 (59%). As indicated by enzyme-linked immunosorbent assay absorbance values, P5-I5414 reached higher virus titers in canola than P5-II5509, which, in turn, reached higher titers than P5-III5594. P5-I5414 was also more virulent in canola than P5-II5509 and P5-III5594, inducing more severe foliar symptoms, stunting, and, in one of two experiments, seed yield loss. Results from this study compared to those of previous studies suggest that analysis of ORF 5 alone is insufficient to assign isolates to coherent strain categories, and further sequencing and phenotyping of field isolates is required.

INVASION AND SUPERINFECTION IN DETERMINISTIC AND STOCHASTIC TWO-STRAIN DENGUE MODELS WITH DEMOGRAPHIC AND SEASONAL VARIATION

The effects of seasonality on disease invasion and superinfection are investigated in dengue epidemic models with two viral strains. A two-strain host–vector nonautonomous ODE is formulated with seasonal periodicity in either vector recruitment or transmission. The basic reproduction number is derived and the dynamics of the single-strain periodic subsystem are summarized. Conditions for uniform persistence and existence of a single-strain periodic solution are verified and the invasion reproduction number is derived. A time-nonhomogeneous, continuous-time Markov chain (CTMC) is formulated from the ODE framework. The basic and invasion reproduction numbers apply to the ODE and CTMC. In the case of invasion, after one strain is established and the invasion reproduction number exceeds one, the ODE predicts a successful invasion, but in the CTMC, there is a periodic probability of invasion that depends on time and size of the invasion. A multitype branching process approximation of the CTMC provides an analytical method to predict the periodic probability of invasion. The numerical results show that the peak time of invasion is related to the current endemic strain and the dominant seasonal driver but it often precedes the peak time of the seasonal driver.

Host Derivation of Sindbis Virus Influences Mammalian Type I Interferon Response to Infection.

Arboviruses are defined by their ability to replicate in both mosquito vectors and mammalian hosts. There is good evidence that arboviruses "prime" their progeny for infection of the next host, such as via differential glycosylation of their outer glycoproteins or packaging of host ribosomal subunits. We and others have previously shown that mosquito-derived viruses more efficiently infect mammalian cells than mammalian-derived viruses. These observations are consistent with arboviruses acquiring host-specific adaptations, and we hypothesized that a virus derived from either the mammalian host or mosquito vector elicits different responses when infecting the mammalian host. Here, we perform an RNA-sequencing analysis of the transcriptional response of Human Embryonic Kidney 293 (HEK-293) cells to infection with either mosquito (Aedes albopictus, C7/10)- or mammalian (Baby Hamster Kidney, BHK-21)-derived Sindbis virus (SINV). We show that the C7/10-derived virus infection leads to a more robust transcriptional response in HEK-293s compared to infection with the BHK-derived virus. Surprisingly, despite more efficient infection, we found an increase in interferon-β (IFN-β) and interferon-stimulated gene (ISG) transcripts in response to the C7/10-derived virus infection versus the BHK-derived virus infection. However, translation of interferon-stimulated genes was lower in HEK-293s infected with the C7/10-derived virus, starkly contrasting with the transcriptional response. This inhibition of ISG translation is reflective of a more rapid overall shut-off of host cell translation following infection with the C7/10-derived virus. Finally, we show that the C7/10-derived virus infection of HEK-293 cells leads to elevated levels of phosphorylated eukaryotic translation elongation factor-2 (eEF2), identifying a potential mechanism leading to the more rapid shut-off of host translation. We postulate that the rapid shut-off of host translation in mammalian cells infected with the mosquito-derived virus acts to counter the IFN-β-stimulated transcriptional response.

Longitudinal map of transcriptome changes in the Lyme pathogen Borrelia burgdorferi during tick-borne transmission

Borrelia burgdorferi (Bb), the causative agent of Lyme disease, adapts to vastly different environments as it cycles between tick vector and vertebrate host. During a tick bloodmeal, Bb alters its gene expression to prepare for vertebrate infection; however, the full range of transcriptional changes that occur over several days inside of the tick are technically challenging to capture. We developed an experimental approach to enrich Bb cells to longitudinally define their global transcriptomic landscape inside nymphal Ixodes scapularis ticks during a transmitting bloodmeal. We identified 192 Bb genes that substantially change expression over the course of the bloodmeal from 1 to 4 days after host attachment. The majority of upregulated genes encode proteins found at the cell envelope or proteins of unknown function, including 45 outer surface lipoproteins embedded in the unusual protein-rich coat of Bb. As these proteins may facilitate Bb interactions with the host, we utilized mass spectrometry to identify candidate tick proteins that physically associate with Bb. The Bb enrichment methodology along with the ex vivo Bb transcriptomes and candidate tick interacting proteins presented here provide a resource to facilitate investigations into key determinants of Bb priming and transmission during the tick stage of its unique transmission cycle.

Longitudinal map of transcriptome changes in the Lyme pathogen Borrelia burgdorferi during tick-borne transmission.

Borrelia burgdorferi (Bb), the causative agent of Lyme disease, adapts to vastly different environments as it cycles between tick vector and vertebrate host. During a tick bloodmeal, Bb alters its gene expression to prepare for vertebrate infection; however, the full range of transcriptional changes that occur over several days inside of the tick are technically challenging to capture. We developed an experimental approach to enrich Bb cells to longitudinally define their global transcriptomic landscape inside nymphal Ixodes scapularis ticks during a transmitting bloodmeal. We identified 192 Bb genes that substantially change expression over the course of the bloodmeal from 1 to 4 days after host attachment. The majority of upregulated genes encode proteins found at the cell envelope or proteins of unknown function, including 45 outer surface lipoproteins embedded in the unusual protein-rich coat of Bb. As these proteins may facilitate Bb interactions with the host, we utilized mass spectrometry to identify candidate tick proteins that physically associate with Bb. The Bb enrichment methodology along with the ex vivo Bb transcriptomes and candidate tick interacting proteins presented here provide a resource to facilitate investigations into key determinants of Bb priming and transmission during the tick stage of its unique transmission cycle.

Rickettsial antibodies and Rickettsia bellii detection in lagomorphs and their ectoparasites in Northern Baja California, Mexico.

Lagomorphs-principally rabbits and hares-have been implicated as hosts for vectors and reservoirs for pathogens associated with multiple rickettsial diseases. Western North America is home to diverse rickettsial pathogens which circulate among multiple wild and domestic hosts and tick and flea vectors. The purpose of this study was to assess lagomorphs and their ectoparasites in 2 locations in northern Baja California, Mexico, for exposure to and infection with rickettsial organisms. In total, 55 desert cottontail rabbits (Sylvilagus audubonii) (Baird) and 2 black-tailed jackrabbits (Lepus californicus) (Gray) were captured. In Mexicali, ticks were collected from 44% (14/32) of individuals, and were exclusively Haemaphysalis leporispalustrisNeumann (Acari: Ixodidae); in Ensenada, ticks were collected from 70% (16/23) individuals, and 95% were Dermacentor parumapertus. Euhoplopsyllus glacialis affinisBaker (Siphonaptera: Pulicidae) fleas were collected from 72% of rabbits and 1 jackrabbit from Mexicali, while the few fleas found on hosts in Ensenada were Echidnophaga gallinaceaWestwood (Siphonaptera: Pulicidae) and Cediopsylla inaequalis(Siphonaptera: Pulicidae). Rickettsia bellii was the only rickettsial organism detected and was identified in 88% of D. parumapertus and 67% of H. leporispalustris ticks from Ensenada. A single tissue sample from a jackrabbit was positive for R. belli (Rickettsiales: Rickettsiaceae). Hosts in Ensenada had a significantly higher prevalence of rickettsial antibodies than hosts in Mexicali (52.3% vs. 21.4%). Although R. bellii is not regarded as pathogenic in humans or other mammals, it may contribute to immunity to other rickettsiae. The marked difference in distribution of ticks, fleas, and rickettsial exposure between the 2 locations suggests that disease transmission risk may vary markedly between communities within the same region.

Creating a Vaccine-like Supplement against Respiratory Infection Using Recombinant Bacillus subtilis Spores Expressing SARS-CoV-2 Spike Protein with Natural Products.

Vaccination is the most effective method of combating COVID-19 infection, but people with a psychological fear of needles and side effects are hesitant to receive the current vaccination, and alternative delivery methods may help. Bacillus subtilis, a harmless intestinal commensal, has recently earned a strong reputation as a vaccine production host and delivery vector, with advantages such as low cost, safety for human consumption, and straightforward oral administration. In this study, we have succeeded generating "S spores" by engineering B. subtilis with spore coat proteins resembling the spike (S) protein of the ancestral SARS-CoV-2 coronavirus. With the addition of two immunostimulating natural products as adjuvants, namely Astragalus membranaceus (Fisch.) Bge (AM) and Coriolus versicolor (CV), oral administration of S spores could elicit mild immune responses against COVID-19 infection without toxicity. Mucosal IgA against the S protein was enhanced by co-feeding with AM and CV in an S spores-inoculated mouse model. Faster and stronger IgG responses against the S protein were observed when the mice were fed with S spores prior to vaccination with the commercial COVID-19 vaccine CoronaVac. In vitro studies demonstrated that AM, CV, and B. subtilis spores could dose-dependently activate both macrophages and dendritic cells by secreting innate immunity-related IL-1β, IL-6, and TNF-α, and some other proinflammatory chemokines and cytokines. In conclusion, the combination of S spores with AM and CV may be helpful in developing a vaccine-like supplement against respiratory infection.

Evolutionarily diverse origins of deformed wing viruses in western honey bees

Novel transmission routes can allow infectious diseases to spread, often with devastating consequences. Ectoparasitic varroa mites vector a diversity of RNA viruses, having switched hosts from the eastern to western honey bees (Apis cerana to Apis mellifera). They provide an opportunity to explore how novel transmission routes shape disease epidemiology. As the principal driver of the spread of deformed wing viruses (mainly DWV-A and DWV-B), varroa infestation has also driven global honey bee health declines. The more virulent DWV-B strain has been replacing the original DWV-A strain in many regions over the past two decades. Yet, how these viruses originated and spread remains poorly understood. Here, we use a phylogeographic analysis based on whole-genome data to reconstruct the origins and demography of DWV spread. We found that, rather than reemerging in western honey bees after varroa switched hosts, as suggested by previous work, DWV-A most likely originated in East Asia and spread in the mid-20th century. It also showed a massive population size expansion following the varroa host switch. By contrast, DWV-B was most likely acquired more recently from a source outside East Asia and appears absent from the original varroa host. These results highlight the dynamic nature of viral adaptation, whereby a vector's host switch can give rise to competing and increasingly virulent disease pandemics. The evolutionary novelty and rapid global spread of these host-virus interactions, together with observed spillover into other species, illustrate how increasing globalization poses urgent threats to biodiversity and food security.

Design and Analysis of a Processing-in-DIMM Join Algorithm: A Case Study with UPMEM DIMMs

Modern dual in-line memory modules (DIMMs) support processing-in-memory (PIM) by implementing in-DIMM processors (IDPs) located near memory banks. PIM can greatly accelerate in-memory join, whose performance is frequently bounded by main-memory accesses, by offloading the operations of join from host central processing units (CPUs) to the IDPs. As real PIM hardware has not been available until very recently, the prior PIM-assisted join algorithms have relied on PIM hardware simulators which assume fast shared memory between the IDPs and fast inter-IDP communication; however, on commodity PIM-enabled DIMMs, the IDPs do not share memory and demand the CPUs to mediate inter-IDP communication. Such discrepancies in the architectural characteristics make the prior studies incompatible with the DIMMs. Thus, to exploit the high potential of PIM on commodity PIM-enabled DIMMs, we need a new join algorithm designed and optimized for the DIMMs and their architectural characteristics. In this paper, we design and analyze Processing-In-DIMM Join (PID-Join), a fast in-memory join algorithm which exploits UPMEM DIMMs, currently the only publicly-available PIM-enabled DIMMs. The DIMMs impose several key challenges on efficient acceleration of join including the shared-nothing nature and limited compute capabilities of the IDPs, the lack of hardware support for fast inter-IDP communication, and the slow IDP-wise data transfers between the IDPs and the main memory. PID-Join overcomes the challenges by prototyping and evaluating hash, sort-merge, and nested-loop algorithms optimized for the IDPs, enabling fast inter-IDP communication using host CPU cache streaming and vector instructions, and facilitating fast rank-wise data transfers between the IDPs and the main memory. Our evaluation using a real system equipped with eight UPMEM DIMMs and 1,024 IDPs shows that PID-Join greatly improves the performance of in-memory join over various CPU-based in-memory join algorithms.

Extracellular vesicles secreted by Brugia malayi microfilariae modulate the melanization pathway in the mosquito host

Vector-borne, filarial nematode diseases cause significant disease burdens in humans and domestic animals worldwide. Although there is strong direct evidence of parasite-driven immunomodulation of mammalian host responses, there is less evidence of parasite immunomodulation of the vector host. We have previously reported that all life stages of Brugia malayi, a filarial nematode and causative agent of Lymphatic filariasis, secrete extracellular vesicles (EVs). Here we investigate the immunomodulatory effects of microfilariae-derived EVs on the vector host Aedes aegypti. RNA-seq analysis of an Ae. aegypti cell line treated with B. malayi microfilariae EVs showed differential expression of both mRNAs and miRNAs. AAEL002590, an Ae. aegypti gene encoding a serine protease, was shown to be downregulated when cells were treated with biologically relevant EV concentrations in vitro. Injection of adult female mosquitoes with biologically relevant concentrations of EVs validated these results in vivo, recapitulating the downregulation of AAEL002590 transcript. This gene was predicted to be involved in the mosquito phenoloxidase (PO) cascade leading to the canonical melanization response and correspondingly, both suppression of this gene using RNAi and parasite EV treatment reduced PO activity in vivo. Our data indicate that parasite-derived EVs interfere with critical immune responses in the vector host, including melanization.

Seasonal Variation and Factors Affecting Trypanosoma theileri Infection in Wild Sika Deer (Ezo Sika Deer Cervus nippon yesoensis) in Eastern Hokkaido.

Trypanosoma (Megatrypanum) spp. are isolated from domestic and wild ruminants, including deer, worldwide. The prevalence of trypanosomes in mammals is influenced by a number of factors such as host age and vector abundance. However, the seasonal variation of and factors affecting trypanosome infection in the wild deer population remain elusive. In this study, we analyzed the seasonal variation in trypanosome prevalence and the factors that affect Trypanosoma theileri Laveran, 1902, infection in wild sika deer (Ezo sika deer) Cervus nippon yesoensis (Heude, 1884) in Eastern Hokkaido through a two-year survey. Seasonal variation in the prevalence of trypanosome infection in the deer population ranged from 0 to 41% as per hematocrit concentration and 17 to 89% as per PCR results. In general, the prevalence of T. theileri by PCR in 2020 was higher than that in 2019. Moreover, the prevalence was significantly higher in the aged population than among the younger population. These findings may explain why individual conditions and sampling season were associated with trypanosome prevalence. This is the first study to investigate the seasonal variation in and risk factors affecting trypanosome infection in wild deer.

Trypanosomatids in Bloodsucking Diptera Insects (Ceratopogonidae and Simuliidae) Wild-Caught at Raptor Bird Nests in Temperate Forests

Trypanosomatids are a huge group of vertebrate, invertebrate and plant parasites that can cause severe illnesses in hosts. Although avian trypanosomes are widely spread throughout the world, information about their transmission and vector–host relationships is still scarce. This study aimed to investigate Trypanosoma prevalence in bloodsucking Diptera collected at tree-nesting raptor birds’ nests. Insects were collected in temperate forests of Eastern-Central Europe, in Lithuania, using UV light traps hung near common buzzard (Buteo buteo) and lesser spotted eagle (Clanga pomarina) nests at about 15 m height above the ground. A total of 1248 Culicoides (Ceratopogonidae) females and 3 blackflies (Simuliidae) were collected and tested for the presence of trypanosomatids using PCR-based methods. The blood of 85 nestlings, belonging to three different species (Buteo buteo, Clanga pomarina and Haliaeetus albicilla) was collected and tested using the same methods. We found that 11.1% of the investigated insects (one Simulium female and Culicoides biting midges belonging to five species) were infected with Trypanosoma parasites (Trypanosoma sp., T. bennetti group, T. avium) and monoxenous trypanosomatids (Crithidia sp., Obscuromonas sp.). Only one common buzzard nestling was found to be infected with Trypanosoma avium. The phylogenetic relationships of detected parasites were determined. Our findings supplement information on the ornithophilic behavior of Culicoides females, potential vector species of avian Trypanosoma, and produce some new information on the detection of monoxenous trypanosomatids (Crithidia sp. and Obscuromonas sp.) in Culicoides.

Sex-typing of ingested human blood meal in Anopheles stephensi mosquito based on the amelogenin gene.

Identifying the sex of human hosts of insect disease vectors, using PCR amplification of the amelogenin gene (AMEL) from the ingested blood meal is an increasingly useful technique for epidemiological studies of vector-borne diseases, as well as within the criminal justice system. Detection of DNA from ingested blood is influenced by the choice of DNA extraction method, genomic target region, type and length of PCR, and rate of degradation in the DNA samples over time. Here, we have tested two types of PCR (i.e. conventional and nested), producing differently-sized PCR products, in time-course assays targeting the human AMEL gene in Anopheles stephensi mosquitoes that were fed on human male and female blood. The fed female mosquitoes were allowed to digest at 28°C for times ranging from 0 to 120h. Three AMEL primer pairs were used to amplify three sequences that were 977, 539, and 106 bp for the X chromosome and 790, 355, and 112 bp for Y. We found that time since feeding had a significant negative effect on the success of PCR amplification. The shortest fragments (106 and 112 bp) were amplified for the longest time after blood feeding (up to 60h), whereas the medium and longest loci were not amplified by conventional PCR even at 0h. However, the nested PCR protocol, targeting the medium sequence, could detect small amounts of human DNA up to 36h (1.5 days) after the blood meal. The shortest PCR assay standardized herein successfully detected small amounts of human DNA in female mosquitoes up to 60h after the blood meal. This assay represents a promising tool for identifying the sex of the human host from the blood meal in field-collected female mosquitoes.