Acta Tropica

A field trial was conducted using 10% lambda-cyhalothrin microcapsule suspension to provide a method for killing ticks and preventing diseases in outdoor gatherings of people or temporary resettlement places after disasters. In this study, three field experimental sites were selected, and each experimental site was set up with a test area and a control area. Before pesticide application, the tick density in three test areas and three control areas was surveyed using the flagging method. Subsequently, two methods were used for pesticide spraying: motorized fogging and electric constant-volume spraying (with the pesticide diluted 300 times). The relative density decline rate of ticks was calculated in three test sites on days 1, 7, 14, 21, and 28 after spraying, and all experimental areas achieved good tick-killing effects. Even without prohibiting wild animals, grazing sheep, and dogs (which are often infested with ticks and not treated) from entering the trial sites, spraying 10% lambda-cyhalothrin microcapsule suspension could maintain a tick-free (low-density) state for approximately 3-4 weeks. Our study provides an idea for controlling epidemics through tick elimination during the high incidence period of tick-borne diseases.

Dengue transmission is strongly influenced by climatic conditions that affect mosquito population dynamics and virus circulation. In Southern Brazil, where dengue historically occurred at low levels, recent climatic anomalies may be contributing to the expansion of Aedes vectors and an increase in local dengue incidence. This study investigated the spatiotemporal association between climatic variables, Aedes mosquito infestation and dengue cases in Porto Alegre (Southern Brazil, 2018 to 2025). Entomological, surveillance and climatic data were analyzed using Morans I and LISA for spatial association, Kendall correlation, polynomial regression and LASSO to identify relevant drivers and develop predictive models of mosquito infestation and dengue incidence. A strong spatial association between Aedes aegypti and Aedes albopictus was observed, with persistent local clusters detected across all years. Annual climatic variables were associated with mosquito abundance in several districts. Overall, rainfall frequency had a stronger effect on Aedes aegypti abundance than accumulated rainfall. Temperature and lagged infestation indices showed strong association with both species and dengue incidence, with effects observed up to four weeks prior. Predictive models demonstrated good agreement between observed and predicted values, particularly within low to moderate infestation levels. Lagged variables were consistently retained in both mosquito infestation abundance and dengue incidence models, highlighting the importance of temporal predictors for anticipating vector dynamics and dengue risk. This approach is generally applicable for predicting Aedes infestation and disease incidence and emphasizes the importance of integrating entomological and climatic surveillance data to improve anticipation and detection of dengue risk periods and support more effective public health interventions.

Some Anopheles species that act as malaria vectors are members of species complexes, a concept whereby sibling species cannot be differentiated solely on the basis of morphological characters. Therefore, species complexes represent a major problem in malaria vector control, because within an Anopheles complex, vectors cannot be differentiated from non-vector species, unless molecular techniques are used to identify them. The Anopheles tessellatus species complex is an important potential vector in South, East, and Southeast Asia, including certain regions of Indonesia. However, no in-depth studies have been conducted on this species complex in that country. Therefore, this study investigated the taxonomic status of An. tessellatus from diverse populations across five Indonesian islands (Sumatra, Java, West Nusa Tenggara, East Nusa Tenggara, and Sulawesi) and identified interpopulation genetic variation based on molecular data of the ITS2, COI, and COII genes. Phylogenetic relationships were constructed using the Maximum Likelihood method. Haplotype and network analysis were also conducted. The results indicate that An. tessellatus constitutes a monophyletic group comprising three well-defined lineages that exhibit clear intraspecific genetic differentiation. Cluster 1 corresponds to the population of Sumatra, Cluster 2 represents population from Sulawesi, and Cluster 3 encompasses populations from Java, West Nusa Tenggara, and East Nusa Tenggara. These findings demonstrate high haplotype diversity and low nucleotide diversity within the species. Populations from West Sumatra, Manado, Tojo Una - Una, and North Morowali (Sulawesi) have the potential for speciation with a genetic distance of 0.61 - 0.94% for COI, between 0.81 - 0.95% for ITS2, and between 0.62 - 0.71% for COII. These findings underscore the need for further integrative studies to obtain a more comprehensive understanding of the An. tessellatus complex in Indonesia and its role in malaria transmission.

PurposeAnopheles stephensi is a malaria mosquito vector that has been raising international concern due to its invasive nature in Africa, including the nation of Djibouti. Since its initial detection in Djibouti in 2012, malaria morbidity and mortality have increased exponentially in the county. While there is an observed association increase in human malaria cases since the arrival of An. stephensi, high-quality evidence of An. stephensi carrying infective sporozoites is essential to determine the role of the invasive vector in malaria dynamics in Djibouti. This study seeks to confirm the link between An. stephensi and malaria transmission in Djibouti and examine genetic relatedness between Djiboutian An. stephensi populations and populations across the Horn of Africa. Such information regarding the An. stephensi populations and the Plasmodium species they transmit is necessary to devise appropriate control strategies and limit malaria transmission within and beyond the country. MethodsOne hundred and ninety-six adult An. stephensi mosquitoes from Djibouti were collected, molecularly confirmed, analyzed for a portion of the cytochrome c oxidase subunit 1 (COI), and tested for infective sporozoites using a highly sensitive and specific multiplex circumsporozoite enzyme linked immunosorbent assay (csELISA) bead assay. The COI sequences of one hundred and fourteen samples were further used to characterize the population genetic structure of the sampled An. stephensi and its genetic relatedness to other An. stephensi populations across the Horn of Africa. ResultsAll 196 samples were morphologically and molecularly confirmed to be An. stephensi. Plasmodium vivax210 sporozoites were detected with a positivity rate of 1.02%. An analysis of the COI region showed that the infected An. stephensi have the most prevalent COI haplotypes of invasive An. stephensi circulating in the Horn of Africa. ConclusionsThe findings from this study confirm the involvement of An. stephensi in P. vivax transmission in Djibouti and describe the genetic relatedness of Djiboutian An. stephensi populations to other populations across the Horn of Africa. This highlights the threat of An. stephensi invasion and supports a rapid and comprehensive response to mitigate the harm that An. stephensi populations cause, particularly through surveillance and control of adult populations. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=119 SRC="FIGDIR/small/707780v1_ufig1.gif" ALT="Figure 1"> View larger version (22K): org.highwire.dtl.DTLVardef@19f762corg.highwire.dtl.DTLVardef@7624forg.highwire.dtl.DTLVardef@c7492borg.highwire.dtl.DTLVardef@194c11f_HPS_FORMAT_FIGEXP M_FIG C_FIG

BackgroundDengue fever is a major neglected tropical disease with a rapidly rising global burden, and localized outbreaks are increasingly reported in southern subtropical China. Fujian Province, a coastal subtropical region with favorable ecological conditions for Aedes albopictus breeding and frequent cross-border exchanges with dengue-endemic areas, has had continuous local dengue cases for over a decade, raising concerns about the establishment of a stable natural endemic focus. Sustained local dengue transmission is defined by four core criteria, but no systematic assessment of these criteria has been conducted for Fujian using long-term multi-dimensional surveillance data. We aimed to evaluate whether a natural endemic focus for sustained local dengue transmission has been established in Fujian Province from 2014 to 2024 using four core evidence dimensions. MethodsWe extracted data on imported and locally acquired dengue cases in Fujian from 2014 to 2024 from Chinas National Notifiable Disease Reporting System (NNDRS). Serological surveillance for dengue IgG antibodies and virological surveillance for dengue virus in Aedes albopictus were conducted at seven sentinel sites. The study period was stratified into three phases based on the impact of COVID-19 non-pharmacological interventions: pre-pandemic (2014-2019), pandemic(2020-2022), and post-pandemic(2023-2024). Descriptive epidemiological analysis and data visualization were performed using R software (version 4.4.1), with t-tests for continuous variables and {chi}{superscript 2} tests for categorical variables. ResultsA total of 3,606 dengue cases were reported in Fujian during the study period, including 1,229 imported and 2,377 locally acquired cases. Key findings were as follows: (1) Temporal distribution: Local dengue transmission was completely interrupted during the 2020-2022 COVID-19 pandemic (0 local cases, only 26 imported cases), and resumed at a low level in 2023-2024 (160 local cases). (2) Serology: The overall population dengue IgG antibody positivity rate was 4.2% (66/15,736), with no statistically significant difference between pre-epidemic (3.8%, 30/7,835) and post-epidemic seasons (4.5%, 36/7,901; P=0.48), and no year with a positivity rate exceeding 10%. (3) Vector surveillance: Only one dengue virus-positive sample was detected among 385,000 Aedes albopictus mosquitoes collected during routine surveillance (Taijiang District, Fuzhou, October 2017), with no viral nucleic acid detected in all other samples. (4) Age distribution: The mean age of locally acquired cases (46.1{+/-}19.8 years) was significantly higher than that of imported cases (35.8{+/-}11.2 years, P<0.001), and local cases were concentrated in the middle-aged group (40-60 years) with no child-dominant pattern observed. ConclusionsFujian Province has not established a stable natural endemic focus for sustained local dengue transmission, and imported cases are the primary driver of local outbreaks in the region. Strengthened surveillance and early management of imported cases, integrated vector control targeting Aedes albopictus, and targeted public health education are critical and essential strategies to prevent the establishment of a dengue natural endemic focus in Fujian and other subtropical coastal regions with similar epidemiological characteristics. Author SummaryDengue fever is a rapidly spreading neglected tropical disease worldwide, and southern China faces persistent threats of local transmission due to favorable ecological conditions for mosquito breeding and frequent cross-border travel. Fujian Province, a subtropical coastal region in southeastern China, has reported annual local dengue cases for over a decade, raising public health concerns about the potential establishment of a stable natural endemic focus--where the virus circulates sustainably without relying on imported cases. To address this critical question, we conducted a comprehensive 11-year assessment (2014-2024) of dengue transmission in Fujian using four key evidence dimensions defined for identifying dengue endemic foci: the continuity of local cases independent of imported sources, population antibody levels, dengue virus detection in local mosquitoes (Aedes albopictus), and the age distribution of infected patients. We also leveraged the COVID-19 pandemic(2020-2022) as a unique natural experiment, during which strict travel restrictions drastically reduced imported dengue cases, to test whether local transmission could persist on its own. Our findings showed that local dengue transmission in Fujian completely stopped during the COVID-19 pandemic and only resumed when cross-border travel and imported cases recovered, confirming local transmission is entirely dependent on imported virus sources. Additionally, the local population had a very low dengue antibody positivity rate (4.2%), dengue virus was detected in only one mosquito sample over 11 years of surveillance, and local cases were concentrated in middle-aged adults (not children--the typical group affected in endemic areas). Together, these results confirm that Fujian Province has not established a stable natural endemic focus for dengue fever. While no endemic focus exists yet, Fujian remains at high risk of imported-driven local outbreaks due to its climate and cross-border exchanges. Our study highlights three critical strategies to prevent the future establishment of a dengue endemic focus in Fujian and other similar subtropical coastal regions: strengthening surveillance and early response for imported dengue cases, implementing targeted mosquito control measures during peak transmission seasons, and conducting public health education to raise awareness of dengue prevention. These evidence-based interventions are key to blocking the formation of sustained local dengue transmission and protecting regional population health.

Urbanization in the 21st century has given rise to complex socio-environmental landscapes that contribute to spatial inequalities in health, particularly in the context of emerging infectious diseases, such as dengue fever. This study employs an urban systems framework to explore the multi-dimensional drivers of dengue risk in Ibague, Colombia, where Aedes mosquitoes thrive in diverse urban environments. We characterized the biophysical, socio-economic, and institutional properties of the urban landscape and employed hierarchical cluster analysis to define urban typologies at both census block and urban section levels. Our findings reveal significant differences in dengue incidence across these typologies, with higher rates associated with areas of high population density and commercial activity. Additionally, we examined the landscape configuration and its role in shaping dengue risk, identifying that diversity and intermixing of typologies had protective effects against dengue incidence. This research underscores the importance of considering multi-scale, socio-ecological factors in dengue risk assessments and highlights the need for targeted public health interventions that address the complex interactions within urban landscapes.

Tick-borne pathogens are transmitted by tick bites to cause infectious diseases in humans and domestic animals. To anticipate the occurrence of tick-borne diseases, it is required to understand high resolution distribution of infection risk and associated ecological factors. The aim of the present study is to reveal the spatial distribution of ticks and ticks infected with pathogens in central Hokkaido, Japan. Adult and nymphal ticks were collected with a constant effort at 171 sites from 13 May to 26 June 2024, followed by screening tick-borne pathogens. The potential suitable habitats of seven tick species and the endemic tick-borne pathogens in the study area (i.e. tick-borne encephalitis virus, Yezo virus, Beiji nairovirus, Lyme disease group borreliae, and relapsing fever group borreliae) were predicted using ecological niche modeling. Ixodes persulcatus and Ixodes ovatus were identified as the primary ticks to determine the distributions of all the pathogens. Besides, the predicted suitable habitats were specific to each pathogen/tick species. Among the environmental variables considered for modeling, snow depth appeared to significantly contribute to the distribution differences between ticks and pathogens. The findings of this study expand our understanding of the spatial risk distribution of tick-borne pathogen infections and its ecological context.

Anopheles albimanus (Nyssorhynchus) is featured as the main malaria vector on Hispaniola. However, five other Anopheles species have been reported circulating in the area; four of them belonging to the subgenus Anopheles (An. crucians, An. grabhamii, An. pseudopunctipennis, and An. vestitipennis) and another one to the Nyssorhynchus subgenus (An. argyritarsis). Previous studies on mosquitoes in the genus Anopheles have identified and characterized peptides from immunogenic salivary proteins, with several of these peptides being unique to the Nyssorhynchus and Anopheles subgenera. This underscores their potential use as biomarkers for differentiating exposure to Anopheles mosquitoes in both the Old World and New World. Since both Nyssorhynchus and Anopheles subgenera have been reported in Haiti, a series of ELISAs were conducted to quantify IgG antibody titers against three published antigenic anopheline salivary peptides (gSG6-P1, Peroxi-P3, and Apy-2) in 348 participants registered in Haitis multi-partner/multidisciplinary Malaria Zero Program. This study aimed to evaluate the intensity of human-vector contact with Anopheles from both subgenera in Grand Anse, Haiti. In addition, the study measured antibodies against a panel of Plasmodium falciparum antigens to determine any association between anti-parasite and anti-peptide antibodies. Significantly elevated IgG responses to Peroxi-P3 in comparison to Apy2 and gSG6-P1 in the total study population (p < 0.001) were observed. Additionally, immune responses to Peroxi-P3 and gSG6-P1 differed significantly between [&le;]18-year-olds and >18-year-olds (p = 0.004 and p = 0.002), whereas no sex-based differences were observed for any peptide. Correlation analyses observed a greater number of significant positive associations in immune response between gSG6-P1 and Plasmodium antigens than any other salivary peptide, an occurrence which was more pronounced in [&le;]18-year-olds than >18-year-olds. A marked reduction in IgG responses to Apy2 and Peroxi-P3, but not gSG6-P1, among participants who kept a single household animal species compared with those who owned two or more species or those who did not have household animals was also demonstrated. Spatial analysis revealed heterogenous geographic overlap of high antibody responses among Peroxi-P3, Apy2, and gSG6-P1, alongside geographically overlapping clusters of low antibody responses to Peroxi-P3 and Apy2. These results provide additional data on the utility of anopheline salivary peptides to characterize human-vector-parasite exposure dynamics in low-transmission areas, such as Haiti.

BackgroundTegumentary leishmaniasis (TL) caused by Leishmania infantum has re-emerged in northern Italy, raising questions about the genetic diversity and population structure of circulating parasites and their potential role in shaping different clinical outcomes. Methodology/Principal findingsMultilocus microsatellite typing (MLMT) based on 15 polymorphic loci was applied to 44 L. infantum strains obtained from TL cases diagnosed between 2013 and 2024 in the Emilia-Romagna region. These strains were compared with sympatric isolates from VL cases, dogs and sand flies. MLMT revealed a considerable genetic variation among TL-associated strains, with 43 distinct multilocus genotypes identified. Population structure analyses using Bayesian clustering, multivariate approaches and phylogenetic reconstruction consistently identified three highly differentiated genetic populations (Fst >0.25). TL strains were divided into two main populations: one shared with VL-associated strains (PopB; 9/44) and a second population found exclusively among TL cases (PopC; 28/44). In contrast, the canine-associated population (PopA) showed no overlap with TL cases in this region. Populations also displayed divergent heterozygosity patterns, as indicated by positive and negative Fis values. Conclusions/SignificanceThese findings revealed previously unknown diversity within L. infantum in the study area and demonstrated that inclusion of tegumentary strains is essential to uncover hidden components of parasite population structure. The identification of a TL-associated population supports the existence of multiple evolutionary pathways and emphasises the importance of integrated One Health surveillance, which combines data from humans, animal hosts and vectors to improve understanding of the epidemiology of leishmaniasis in Italy. Author summaryLeishmania infantum is a parasite transmitted to humans through the bite of infected insect vectors. It can cause different forms of leishmaniasis, ranging from a systemic disease known as visceral leishmaniasis to a less common form that affects the skin and mucous membranes, called tegumentary leishmaniasis. Dogs are the main reservoir of the parasite and play a key role in maintaining its circulation in endemic areas. In recent years, cases of tegumentary leishmaniasis have re-emerged in northern Italy. This unexpected increase has raised questions about how the parasite is spreading and whether genetic differences among the parasites could explain these new patterns. To explore this, we examined parasitic DNA obtained from tegumentary leishmaniasis cases and compared it with DNA from patients with visceral leishmaniasis, from dogs and insect vectors from the same area. By examining multiple genetic markers, we found that parasites causing the tegumentary form of the disease are genetically diverse and belong to different groups. Notably, one parasite group was found only in cases of tegumentary leishmaniasis and not in visceral infections nor in infected dogs, suggesting that some parasite lineages may be more closely associated with skin and mucosal disease. Overall, our findings show that studying parasites from cutaneous and mucosal lesions provides important information that would otherwise remain hidden. By combining data from humans, animals and insect vectors, our study highlights the importance of integrated surveillance systems for improving our understanding of disease spread and supporting effective public health strategies.

BackgroundAnopheles mosquitoes vector pathogens responsible for more than 600,000 human deaths annually. Ecological studies of these insects are important to guide effective vector-control campaigns and to understand their broader ecological consequences. Molecular ecology methods, particularly qPCR, provide a valuable tool in such studies. By detecting trace DNA of a taxon of interest within mixed or environmental samples, qPCR can facilitate identification of prey taxa of interest in the diets of consumers. However, no protocol for the detection of An. gambiae complex mosquitoes in dietary samples has been available. MethodsWe introduce a new set of qPCR primers (Agam_CO1_F1 and Agam_CO1_R1) and a probe-based assay for detection of Anopheles gambiae-complex mosquitoes, even with short reads common in dietary and environmental samples. The primers were tested in vitro for their specificity and sensitivity, and in silico using Primer-BLAST to assess potential off-target amplification. ResultsThe qPCR primers amplified An. gambiae DNA even at low starting concentrations (5 copies {micro}l-1). The primers did not amplify any non-target DNA in either the in vitro or in silico tests, but consistently amplified An. gambiae complex DNA. The primers can therefore provide reliable tests for the presence or absence of An. gambiae complex in dietary or eDNA samples. ConclusionsThe new qPCR primers should allow advances in research into mosquito ecology by allowing detection of even trace amounts of An. gambiae DNA in dietary and environmental samples. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=72 SRC="FIGDIR/small/707393v1_ufig1.gif" ALT="Figure 1"> View larger version (11K): org.highwire.dtl.DTLVardef@1fcce3corg.highwire.dtl.DTLVardef@47e5f5org.highwire.dtl.DTLVardef@4a7063org.highwire.dtl.DTLVardef@1188d60_HPS_FORMAT_FIGEXP M_FIG C_FIG

Climate change driven saltwater intrusion is expanding coastal brackish water habitats, promoting the proliferation of salinity-tolerant mosquitoes such as Culex sitiens, a vector of lymphatic filariasis and Japanese encephalitis. This study investigated whether environmental salinity affects mosquito morphology, specifically proboscis length, a trait of ecological significance related to feeding efficiency and vector competence. Late fourth-instar larvae and pupae of Cx. sitiens were collected from habitats with varying salinity levels in the Muttukadu Backwater, Tamil Nadu, India, and reared under controlled conditions using habitat specific water. Adult female body and proboscis lengths were measured, and water quality parameters were analysed to characterize environmental variation. Statistical analyses (Welchs ANOVA, regression, and ANCOVA) revealed a significant positive relationship between salinity and body length (R2 = 0.94, p=0.0003) as well as with proboscis length (R2 = 0.90, p=0.001). Additionally, ANCOVA indicated that the proboscis elongation remained significant after adjusting for body length (F{square}, {square}{square}{square} = 32.36, p < 0.001, partial 2 = 0.257). This confirmed that the salinity exerts an independent effect on this morphological trait. These findings provide the first field-based evidence that the environmental salinity drives proboscis elongation in Cx. sitiens, indicating an adaptive response under saline stress. This may have implications for disease transmission in climate-affected coastal regions.

There is empirical evidence that biophysical factors determine the spatio-temporal distribution of mosquito vectors, and identifying the variables that shape their ecology allows decision-makers to design effective surveillance and control strategies. This study evaluated the spatiotemporal distribution of Aedes albopictus in relation to environmental and biotic variables in the Iguazu Department, Misiones Province, Argentina, within the tri-border region shared with Brazil and Paraguay. Environmental characterization integrated field data and remotely sensed biophysical variables, and vector occurrence was analyzed at micro- and meso-spatial scales using generalized linear mixed models. Eleven sampling sessions were conducted between April 2019 and February 2020 at 81 sites representing urban, periurban, and wild environments. A total of 1,614 Ae. albopictus and 4,358 Ae. aegypti specimens were identified. Rainfall, minimum temperature, exposure days, and land cover were the main predictors of Ae. albopictus presence, showing nonlinear responses to precipitation and vegetation. The selected model explained 67% of the variance. The species exhibited clear spatiotemporal stratification, with periurban and disturbed wild areas functioning as ecotones favorable to its establishment. These findings provide key insights to guide preventive actions and strengthen integrated vector management strategies in the region.

BackgroundThe Hump-nosed pit viper is a recognized but neglected medically significant species causing morbidity and mortality, with non-availability of a specific antivenom. There are many gaps in our understanding of its envenomation, including burden, clinical syndrome, complications and management. MethodologyThe study is a retrospective sub analysis of the Prospective VENOMS registry and hospital records of Hump Nosed Pit Viper envenomation from a single tertiary care center in coastal Karnataka from May 2018 to March 2024. Epidemiology, syndrome, complications and treatment strategies have been described. A linear mixed model analysis was conducted to study the effect of different therapeutic interventions in combating venom induced consumptive coagulopathy (VICC) Principal FindingsOf 46 cases, 24 patients had VICC. The most common complications were AKI (21.7%), TMA (10.9%) and stroke (4.4%). Anaphylaxis to ASV (23.9%) was the most common therapeutic complication. Therapeutic interventions included ASV, administration of blood products and therapeutic plasma exchange along with supportive care. The linear mixed model revealed that administration of blood products (p=<0.001) had the strongest influence on the INR value, however, often resulting in a transient decline in INR value. ASV (p=0.052) caused only marginally significant change in INR. The role of TPE could not be statistically inferred, however, individual cases with severe VICC improved without complications, therefore it required further study but can be considered in critical cases. Conclusions/SignificanceThis study describes the syndrome of hump-nosed pit viper envenomation, while highlighting the urgent need for a species-specific antivenom, recommends treatment strategies that can be used in the interim. Additionally, geo-spatial mapping draws attention to hotspots and the hypothesis that HNPV in coastal Karnataka have regionally distinct toxicity trends. Author SummaryIndia is often known as the snakebite capital of the world, with recent literature suggesting that not all death and disability is attributable to the "Big four" and highlighting regionally significant species of snake. In the Western Ghat region of India, envenomation by the Hump-nosed pit viper is increasingly being reported. In coastal Karnataka, it has been reported as the second most common cause of envenomation following the Russells Viper, causing systemic envenomation and death. However, little is known about why envenomation is common in this region, is it increasing, how to diagnose envenomation, its clinical syndrome, the anticipated complications, and most importantly, an effective treatment strategy. This study reports envenomation in 46 patients, resulting in 3 deaths, and 24 patients developing derangement in coagulation parameters.

Accurate species identification is crucial to assess the medical and veterinary relevance of a mosquito specimen, but it requires high experience of the observers and well-equipped laboratories. This study aimed to evaluate whether low-cost imaging in combination with geometric wing morphometrics can provide accurate identification of invasive, morphologically similar Aedes species. The right wings of 670 female specimens covering 184 Ae. aegypti, 156 Ae. albopictus, 166 Ae. j. japonicus and 164 Ae. koreicus, were removed, mounted and photographed with a professional stereomicroscope (Olympus SZ61, Olympus, Tokyo, Japan) and a macro lens (Apexel-24XMH, Apexel, Shenzhen, China) attached to a smartphone. The coordinates of 18 landmarks on the vein crosses were digitalized by a single observer for each image. In addition, the landmarks of 20 specimens per species and imaging device were digitalized by six different observers to assess the degree of the observer error. The superimposed shape variables were used to compare the species classification accuracy of linear discriminant analysis (LDA), support vector machine (SVM), Random Forest (RF), and XGBoost. In the single-observer landmark data, the LDA achieved the best classification results with a mean accuracy of 95 % for landmarks from microscope images and 92 % for those obtained from smartphone images. In the multi-observer landmark data, LDA consistently performed worse than the other three classifiers, and the reduction in the accuracy was more pronounced for smartphone images than for microscope images. This pattern was associated with a higher degree of observer error for smartphone images, as confirmed by a landmark-wise comparison across all landmarks. Geometric wing morphometrics provides a reliable method to distinguish the most common invasive Aedes species in Europe. Thereby, the image quality obtained by smartphones equipped with a macro lens is sufficient and represents a cost-effective alternative to professional microscopes. However, due to the greater degree of observer variation for smartphone images, landmark coordinates for such images should ideally be collected by a single observer.

Tick-borne relapsing fever (TBRF) is caused by Borrelia species transmitted to humans by soft ticks of the genus Ornithodoros. Very little is currently known about the morbidity of this disease in Mali, despite the risk of human co-infection with malaria. The lack of appropriate diagnostic services or technical expertise to differentiate suspected malaria from other causes of febrile illness of unknown origin means that this tick-borne disease remains neglected and under-diagnosed in febrile patients in Mali. Our study investigated the detection of Borrelia crocidurae DNA in febrile patients in Mali from malaria rapid diagnostic tests (RDTs). MethodsBetween June and December 2021, negative and positive malaria RDTs were collected from 41 sites in the nine regions of Mali. Both qPCR and standard PCR were used to detect the presence of B. crocidurae DNA. ResultsOf the 1496 malaria RDTs tested, B. crocidurae DNA was detected in 9 malaria-negative RDTs. All of these were collected in the region of Kayes, where the prevalence rate was 6% (9/150) of the negative malaria RDTs. ConclusionsThis study demonstrates that tick-borne relapsed fever is an under-diagnosed condition in febrile patients in Mali.

Schistosomiasis is a neglected disease that is still ravaging many rural community dwellers in Nigeria. Different species of freshwater snails, such as Bulinus and Biomphalaria, play passive roles in the transmission of schistosomiasis. A study on the occurrence of snail intermediate hosts of schistosomes was conducted following mass praziquantel administration in Katsina-Ala, Benue State, Nigeria. The objective was to determine the prevalence of freshwater snails. incriminated in transmitting schistosomiasis and their cercaria infectivity. A total of 24,812 freshwater snails belonging to six different genera (Bulinus globosus, B. forskalii, Pila ovata, Lanistes lybicus, Biomphalaria pfeifferi, and Lymnaea natalensis were encountered. Snails collected were kept under laboratory conditions for cercaria emergence. B. globosus, B. forskalii, and B. pfeifferi were identified as vectors of Schistosomes because they secreted cercaria of S. haematobium and S. mansoni, respectively. Parasite infection was higher in the ponds (83.33 %); followed by dams (68.57 %)) and streams (53.33 %). There was positive correlation (r = 0.657; P < 0.05) between the amount of rainfall and abundance of all snail species and their infection rate collected except Biomphalaria species that showed a negative correlation. The studies, therefore, call for Consideration of snails control to be integrated with Praziquantel Mass Drug Advocacy Campaigns so as to achieve transmission interruption.

IntroductionDengue has been prevalent in a regular fashion in Bangladesh and Chattogram for the last 6-7 years and is showing some serotype twisting. So, the objectives of the present study were to explore the burden of dengue serotypes in Chattogram. MethodsIn this study, 223 Dengue RT-PCR positive patients were evaluated for serotyping. Gender and age group, along with cycle threshold (CT) values, were also collected. Data after collection were compiled, analyzed, and plotted in Microsoft Excel and GraphPad Prism 10.4. Ethical clearance was taken to conduct the study. ResultsAmong 223 patients analyzed, males and females were found near equal (113 and 110). Middle-aged patients were more than the extremes of age. The mean {+/-} SD of age was 33.55 {+/-} 13.67 years. Regarding serotype distributions, isolated Den 1, Den 2 and Den 3 were found 1.3%, 73.1% and 6.7%, respectively. Concurrent infections with multiple serotypes were observed in several patients, most notably the Den 2 and Den 3 combination, which accounted for 14.3% (n=32) of the cases. Other co-infections were less frequent: the Den 1 and Den 2 pairing appeared in 3.6% (n=8) of the cohort, while triple-serotype infections (Den 1, 2, and 3) and Den 3/Den 4 pairings were rare, each occurring in only 0.4% of patients. Statistical analysis of CT values revealed no significant sex-based differences for Den 2 and Den 3. However, significant variations in CT values were observed when comparing Den 1 against both Den 2 and Den 3 (p < 0.05). In contrast, the difference between Den 2 and Den 3 Ct values remained statistically insignificant. ConclusionIn the year 2025, Dengue serotypes 2 and 3 were found to be the most prevalent, both in isolated or in combinations and Den 1 and Den 4 were found minimum. Exposure to multiple serotypes and twisting from one serotype to another might influence the dengue outcome in future, which needs further exploration.

BackgroundBoth hemorrhagic fever with renal syndrome (HFRS) and scrub typhus (ST) are acute zoonotic infectious diseases. There is an overlap in their epidemiological characteristics and clinical manifestations, posing challenges for early differential diagnosis. This study aims to identify predictive factors for these two diseases to provide a basis for early diagnosis. Method/FindingsA retrospective analysis was conducted on the clinical data of patients diagnosed with HFRS and ST at the First Affiliated Hospital of Dali University. Logistic regression analysis was employed to explore independent risk factors for the early differential diagnosis of these two diseases, and a nomogram model was constructed based on these risk factors. The performance of the model was evaluated using the area under the receiver operating characteristic curve (AUC). The nomogram was utilized to visually present the predictive variables. Decision curve analysis (DCA) was performed to assess the clinical utility of the model. ResultsA total of 235 patients each with HFRS and ST were included in this study. After adjusting for confounding factors, the results of multivariate logistic regression analysis revealed that sex (male) (adjusted odds ratio [ajOR]: 2.093, 95% confidence interval [CI]: 1.107 - 3.957, P = 0.018), positive proteinuria (ajOR: 4.937, 95% CI: 2.427 - 10.042, P < 0.001), creatinine (CREA) (ajOR: 1.009, 95% CI: 1.003 - 1.015, P = 0.005), heart rate (ajOR: 0.981, 95% CI: 0.966 - 0.997, P = 0.018), and conjunctival congestion (ajOR: 16.167, 95% CI: 5.326 - 49.072, P < 0.001) were independent risk factors for differentiating HFRS from ST. The AUC of the model constructed based on these five independent risk factors was 0.856. ConclusionSex (male), positive proteinuria, elevated CREA, decreased heart rate, and conjunctival congestion are effective predictive factors.

Astyanax Baird & Girard, 1854 is a widely distributed and species-rich genus of Acestrorhamphidae, whose abundant populations in Neotropical basins play a crucial ecological role at the trophic level. Taxonomic uncertainties persist within the genus, as seen in Astyanax sp. (formerly designated as A. fasciatus) from the Magdalena basin in Colombia. Concerns about its genetic status are heightened due to ecological threats posed by hydroelectric dams, from habitat loss to river connectivity. We isolated and characterized 17 microsatellite loci to assess the population genetics of this species in a broad sample from the middle and lower sections of the Cauca River, now interrupted by the Ituango dam. Furthermore, a multidisciplinary approach integrating phylogenetic analyses of mitochondrial (COI) and nuclear (rag2) markers with geometric morphometric analyses was employed to evaluate potential cryptic diversity within Astyanax sp. Microsatellites revealed two genetic groups in the studied area, strongly supported as distinct lineages by phylogenetic analyses. Unexpectedly, one of these lineages of Astyanax sp. was recovered in an unresolved clade with samples of A. microlepis and allopatric samples of A. viejita from the Maracaibo Lake basin. Each genetic group showed high genetic diversity, but also evidence of recent bottleneck events and significant-high values of inbreeding. Morphometric analyses provided evidence of significant phenotypic differentiation among A. microlepis, Astyanax sp. 1 (Asp1), and Astyanax sp. 2 (Asp2). Morphological patterns ranged from the robust profile of A. microlepis to the streamlined shape of Astyanax sp. 2 (Asp2), with Astyanax sp. 1 (Asp1) displaying intermediate traits and localized differences in head length and fin placement. Statistical support from permutation tests and a high overall classification accuracy (95.65%) underscore the existence of distinct morphospecies, suggesting that phenotypic differentiation is well-established, despite the complex evolutionary history of the group. This study suggests the presence of cryptic diversity within Astyanax sp. and provides valuable genetic information for the conservation and management of their populations in the Magdalena basin.

O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=118 SRC="FIGDIR/small/700930v1_ufig1.gif" ALT="Figure 1"> View larger version (63K): org.highwire.dtl.DTLVardef@177a6b4org.highwire.dtl.DTLVardef@6186a3org.highwire.dtl.DTLVardef@ce6196org.highwire.dtl.DTLVardef@168bf43_HPS_FORMAT_FIGEXP M_FIG C_FIG The kangaroo soft tick, Australpavlovskyella gurneyi (Warburton, 1926), is found in sandy depressions ( wallows), under desert shade trees, formed by the activity of the red kangaroo, Osphranter rufus, resting under shade trees (https://youtu.be/AYLoqqPsifc). The field biology of the tick was examined on Moralana Station in arid mid-north, South Australia, between February 1969 and March 1971. The age of kangaroo dung in wallows showed that kangaroos visited wallows regularly during the hot summer and infrequently during the cooler months. All nymphal instars and adults were present at all times of the year in kangaroo wallows, but only a small proportion of the ticks present was trapped on any one occasion. Ticks were abundant in large kangaroo wallows under trees with dense shade, but scarce under smaller trees with sparse shade. The short-lived larvae were present only during spring and early summer, indicating that the long-lived female ticks bred only during spring and early summer. Laboratory tests showed that field-collected adult female ticks entered reproductive diapause from January to August (mid-summer to late-winter). Ticks placed in kangaroo wallows survived for at least one year without food. On Moralana Station, the population of first-instar nymphs increased in summer and subsequently the population of second-instar nymphs increased in early autumn, indicating that a life cycle could be completed in 2-3 years. HighlightsO_LIThe seasonal biology of Australpavlovskyella gurneyi, found in sandy depressions wallows formed by the activity of the red kangaroo, under sparse semi-arid desert shade trees was examined for the first time. C_LIO_LIEngorged ticks placed in kangaroo wallows survived for at least one year without food. C_LIO_LIIn this environment, the entire life cycle could be completed in 2-3 years. C_LI