BACKGROUND:Vector-borne diseases (VBDs), such as malaria, dengue, and Zika virus infections, remain a critical global health burden, particularly in resource-limited regions. Conventional diagnostic methods, including microscopy, enzyme-linked immunosorbent assays, and polymerase chain reaction, face significant limitations due to their reliance on centralized laboratories, lengthy processing times, and high operational costs, hindering timely disease management in remote or underserved areas. OBJECTIVE:Microfluidic diagnostics have emerged as a transformative solution, offering portability, system integration, and high sensitivity through miniaturized fluidic control and multiplexed detection capabilities. These devices enable rapid, on-site diagnosis by combining sample preparation, target amplification, and signal readout into a single chip, aligning with the growing demand for point-of-care testing. METHODS:This review was conducted following a systematic literature search of PubMed, Web of Science, and Scopus databases from January 2010 to June 2026. Search terms included combinations of "microfluidic," "lab-on-a-chip," "point-of-care," "vector-borne disease," "malaria," "dengue," "Zika," "West Nile virus," "Lyme disease," and "CRISPR." Inclusion criteria were (1) peer-reviewed articles reporting microfluidic chip applications for VBD pathogen detection or vector surveillance; (2) studies describing field-deployable or clinically validated platforms; and (3) publications in English. Exclusion criteria included purely theoretical modeling studies without experimental validation and nonchip-based microfluidic systems. A total of 87 studies were selected for qualitative synthesis after title/abstract screening and full-text evaluation. We critically evaluate real-world application scenarios, including field deployments for malaria surveillance in Uganda and outbreak responses to arboviral infections in Brazil and Southeast Asia, while addressing persistent translational barriers such as manufacturing standardization, sample pretreatment complexity, and cost-effectiveness. RESULTS:Key findings demonstrate the successful integration of sample-to-answer workflows on miniaturized platforms, achieving significant reductions in detection time and operational dependencies compared with conventional laboratory methods. Critical challenges identified include manufacturing scalability for resource-limited settings, complex sample pretreatment requirements, and the pressing need for cost reduction to ensure sustainable deployment in endemic regions. CONCLUSION:This review aims to explore emerging integration strategies beyond conventional diagnostics, focusing on approaches such as CRISPR-based assays, artificial intelligence-driven image analysis, and scalable manufacturing processes to strengthen global VBD control networks and improve clinical outcomes in underserved populations. CONCLUSION:This review aims to explore emerging integration strategies beyond conventional diagnostics, focusing on approaches such as CRISPR-based assays, simplified image analysis, and scalable manufacturing processes to strengthen global VBD control networks and improve clinical outcomes in underserved populations. Specific recommendations for phased deployment in endemic regions, including pilot beta-testing programs and technology transfer partnerships, are proposed to accelerate translational impact.
BACKGROUND:Zoonotic malaria caused by simian Plasmodium species presents increasing diagnostic challenges in endemic regions. METHOD:This study developed and evaluated malachite green-based loop-mediated isothermal amplification (MG-LAMP) assays for detecting five zoonotic malaria parasites (P. knowlesi, P. cynomolgi, P. inui, P. coatneyi, and P. fieldi). The assays enable rapid visual detection through a colorimetric change without requiring specialized equipment. To reduce carryover contamination, deoxyuridine triphosphate (dUTP) and uracil-DNA glycosylase (UDG) were incorporated into the reaction. RESULT:All five UDG MG-LAMP assays demonstrated high analytical sensitivity, with limits of detection of 1 copy/µL for P. cynomolgi, P. coatneyi, and P. fieldi, and 10 copies/µL for P. knowlesi and P. inui. The assays showed 100% specificity, with no cross-reactivity among target species. Malachite green enabled clear visual discrimination between positive and negative reactions. CONCLUSION:A simplified direct blood lysis method requiring less than 20 min was also developed to enable rapid DNA preparation without column-based purification. Overall, the MG-LAMP assays combined with UDG and rapid lysis provide a sensitive, specific, and practical approach for diagnosing zoonotic malaria, particularly in resource-limited endemic settings.
BACKGROUND:Tick-borne encephalitis virus (TBEV) is an important cause of neuroinvasive infection in Europe and Asia. Israel is considered nonendemic, yet travel-associated cases have been reported. METHODS:Sera from hospitalized neurological patients in Israel (2010-2019) were screened for TBEV IgG antibodies. Positive/borderline results were confirmed by virus neutralization testing (VNT) against both TBEV and West Nile Virus (WNV). Confirmed TBEV-neutralizing sera were tested for anti-TBEV IgG and IgM by immunofluorescence assay. RESULTS:Of 12,651 suspected meningitis/encephalitis cases, 1,280 (10.1%) were screened. Sixty-three were ELISA-IgG positive/borderline (4.9%), and VNT confirmed TBEV neutralization in 9 (0.7% of the 1,280 patients). West Nile Virus (WNV) Virus neutralization test (VNT) was positive in 51/63 samples. Two samples neutralized both viruses. All TBEV VNT-positive sera were IgM-negative. CONCLUSIONS:Confirmed TBEV exposure was rare with no evidence of missed acute TBE infection. Most apparent TBEV ELISA reactivity reflected WNV cross-reactivity. Parallel VNT against WNV and TBEV is essential in WNV-endemic countries for accurate TBEV diagnosis.
Background: Ticks are the main vectors for a variety of tick-borne bacterial pathogens. Systematic studies on the coinfection patterns and differences of pathogens in ticks derived from different hosts are of great significance. Therefore, the present study investigated ticks collected from multiple host species in the Altun Mountains region (China) to preliminarily characterize differences in the carriage of tick-borne bacterial pathogens across host-associated and environmental tick populations and to explore their potential relationships. Materials and Methods: A total of 2820 ticks collected from marmots, sheep, camels, and Gobi vegetation were assigned into 562 pooled groups. Polymerase chain reaction assays were performed to detect multiple tick-borne bacterial pathogens, followed by phylogenetic analysis of representative sequences. Results: Ticks in the Altun Mountains region carried multiple tick-borne bacterial pathogens with distinct host-associated distribution patterns. Moreover, there were significant differences in the pathogen carriage profiles among ticks from different hosts and environmental sources. In particular, the distributions of spotted fever group (SFG)/typhus group (TG) rickettsiae, Anaplasma spp ., and Coxiella burnetii in ticks from different hosts were significantly different ( p < 0.05). SFG/TG rickettsiae were the dominant pathogens forming highly conserved phylogenetic clusters and widely distributed in ticks. Furthermore, Anaplasma spp. were distributed across multiple phylogenetic clades. Conclusion: These findings indicate that tick-borne pathogens in the Altun Mountains region exhibit distinct host-associated distribution characteristics and complex ecological circulation characteristics.
Background: Global dengue cases have surged in recent years. In Ecuador, reported cases increased from 27,906 in 2023 to 57,712 in 2024. Despite the availability of multiple vaccines in other countries, factors such as socioeconomic disparities, climate change, and vaccine hesitancy continue to hinder prevention efforts. This study aims to analyze the perceptions of the Ecuadorian population and explore possible reasons for hesitancy toward the dengue vaccine. Methods: This cross-sectional observational study was conducted in accordance with the STROBE guidelines. Online surveys, adapted from Kricorian et al., were distributed via the Qualtrics app between June 1st, 2024, and June 25, 2025. Eligible participants were Ecuadorian, at least 18 years old, and had a minimal education level of bachelor. Statistical analysis included descriptive percentages and chi-square tests, with significance set at p < 0.05. Results: A total of 735 participants were included. Most perceived a dengue vaccine as safe (87.2%, n = 641), while 12.8% ( n = 94) considered it unsafe. Overall, 75.9% expressed willingness to be vaccinated. Perceived vaccine safety was significantly associated with vaccination willingness, region of residence, and willingness to vaccinate offspring (all p < 0.001), but not with education level, dengue understanding ( p = 0.747), or prior dengue experience ( p = 0.111). Conclusion: Most Ecuadorians perceive dengue vaccines as safe, yet hesitancy persists. Targeted educational efforts are essential to improve acceptance and coverage.
BACKGROUND:West Nile virus (WNV) is a flavivirus typically transmitted between mosquitoes and birds with occasional spill-over into other animals, including humans. Children evaluated for suspected Lyme disease may share environmental risk factors for WNV and present with overlapping symptoms. Our goal was to determine the prevalence of WNV infection in children with suspected Lyme disease. METHODS:We performed a nested case-control study within a prospective cohort study of children with suspected Lyme disease aged 1-21 years enrolled at 1 of 8 Pedi Lyme Net emergency departments. Lyme disease was defined by either an erythema migrans (EM) lesion or positive two-tier serology. Cases with Lyme disease (n = 305) were matched by age, symptoms, and enrolling center to symptomatic controls without Lyme disease (n = 295). We performed indirect (Luminex microbead immunoassay MIA] with reflex IgM enzyme immunoassay [EIA]) and direct (polymerase chain reaction [PCR]) WNV testing. Acute WNV infection was defined with either a positive MIA and EIA or a positive PCR test result. RESULTS:Of the 600 included patients, 15 (2.5%) had a positive and 53 (8.8%) had an equivocal WNV MIA result. Of these, only 1 had a positive EIA result. Another child had a positive WNV PCR test. The two children with active WNV infections also had acute Lyme disease (0.3%, 95% confidence interval 0.1%, 1.2%) with neurological symptoms. CONCLUSION:Acute WNV infection is uncommon in pediatric patients being evaluated for suspected Lyme disease. Routine WNV testing is not indicated when evaluating for suspected Lyme disease.
BACKGROUND:Urinary tract infections (UTIs) in cats are increasingly recognized as clinically relevant conditions frequently associated with multidrug-resistant (MDR) bacteria of potential zoonotic origin, yet genomic data on feline uropathogens remain scarce in Tunisia. METHODS:We used whole-genome sequencing to characterize seven bacterial isolates recovered from six cats with clinical signs of UTI: Mammaliicoccus lentus (n = 2), Staphylococcus schleiferi (n = 1), Mammaliicoccus sciuri (n = 1), Enterococcus faecalis (n = 1), Enterococcus casseliflavus (n = 1), and Klebsiella aerogenes (n = 1). RESULTS:Resistome analysis revealed determinants conferring resistance to β-lactams (blaZ, blaCMY-132), methicillin (mecC-type), macrolides (erm(43), ermB), tetracyclines (tet(M), tet(45), tetB), fosfomycins (fosI, fosB, fosA5), and aminoglycosides (aac(6'), aph(3')-IIIa, aph(6)-Id), alongside efflux pump genes (efrA, sepA, sdrM, oqxA, KpnE/F/G), vancomycin-operon genes (vanT, vanY, vanC, vanG), and biofilm/biocide-tolerance genes (salB, qacG). Notably, M. lentus S104 carried mecC-type elements, the first such report in Tunisia, while K. aerogenes displayed an extensive MDR profile, including blaCMY-132 and fosA5. Multilocus sequence typing/ribosomal multilocus sequence typing (MLST/rMLST) identified diverse lineages, including the internationally distributed E. faecalis ST19 and the rarely reported K. aerogenes ST242. Plasmids were absent in all isolates; a Tn916/1545-type transposon occurred in E. casseliflavus, and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems were unevenly distributed. CONCLUSIONS:These findings highlight companion animals as reservoirs of clinically important resistance genes, reinforcing the need for One Health AMR surveillance.
BACKGROUND:The Malayan pangolin (Manis javanica) is a critically endangered species that may harbor diverse zoonotic pathogens. This study aimed to characterize tick species parasitizing confiscated Malayan pangolins and determine the prevalence and genetic diversity of spotted fever group rickettsiae (SFGR) associated with these ticks. METHODS:We employed morphological and molecular approaches to identify ticks parasitizing pangolins confiscated in Guangdong Province between 2021 and 2024. A total of 435 adult ticks were collected from 32 pangolins and pooled into 87 groups (5 ticks per pool). Pooled tick samples were screened for SFGR by PCR targeting three rickettsial genes (gltA, ompA, and ompB), followed by Sanger sequencing and phylogenetic analysis. RESULTS:All collected ticks were identified as Amblyomma javanense. The pool positivity rate for SFGR was 41.4% (36/87), with a minimum infection rate of 8.3% (36/435; 95% CI: 5.8-11.2%). Molecular characterization identified the detected agent as Candidatus Rickettsia jingxinensis (Ca. Rickettsia jingxinensis), an agent recently confirmed to infect humans. CONCLUSION:These findings provide baseline data for tick-borne pathogen surveillance in the context of pangolin conservation and illegal wildlife trade control, highlighting the potential public health relevance of the pangolin-tick interface.
BACKGROUND:Leptospirosis is a globally distributed zoonotic disease that remains a significant public health concern, yet integrated evidence linking human infection, animal reservoirs, and Leptospira serogroups remains limited. Pan'an County, a rural sentinel site in eastern China, provides a suitable setting for investigating leptospirosis transmission within a One Health framework. This study aimed to characterize epidemiological patterns in humans and associated animal infections, providing evidence for understanding leptospirosis transmission. METHODS:A retrospective and field-based surveillance study was conducted from 2007 to 2025. Descriptive methods were used to analyze temporal, spatial, and demographic characteristics of human leptospirosis cases. Animal surveillance was conducted across multiple host species to assess infection status and Leptospira serogroup distribution. Correlation analysis was performed to evaluate associations between rodent density, infection status, and human leptospirosis cases. RESULTS:A total of 53 human leptospirosis cases were reported, with a marked peak in 2007 and no fatalities. Cases were concentrated in late summer and primarily affected middle-aged farmers, with no significant spatial clustering observed. Animal surveillance identified 5630 samples with an overall positivity rate of 2.58%. Infections were detected in rodents, frogs, and ducks, while pigs and cattle were negative. Rodents and frogs were exclusively associated with Icterohaemorrhagiae, whereas ducks exhibited multiple serogroups. Rodent density was positively correlated with human cases, while no association was observed between infection positivity rates and human incidence. CONCLUSION:Leptospirosis in Pan'an is characterized by a stable Icterohaemorrhagiae-dominated transmission system primarily maintained by rodent reservoirs, with human infection occurring through occupational and environmental exposure in agricultural settings. Aquatic-associated animals such as frogs and ducks likely function as environmental interface hosts within a One Health transmission framework. These findings suggest a rodent-associated transmission cycle and support the value of integrated human-animal-environment surveillance for leptospirosis control in rural endemic regions.
BACKGROUND:Previous studies have identified routine laboratory abnormalities associated with human granulocytic anaplasmosis (HGA). To aid clinicians in evaluating patients with suspected disease while waiting for polymerase chain reaction (PCR) results, we describe the diagnostic testing characteristics of routine laboratory tests compared with a gold standard PCR. DESIGN:We conducted a retrospective chart review of emergency department patients in a highly endemic region who had an HGA PCR sent as part of their clinical care. We reviewed 223 eligible PCR-positive cases and a randomly sampled selection of 466 eligible PCR-negative cases. We calculated the positive and negative likelihood ratios (LR+ and LR-) for common laboratory abnormalities (leukopenia, thrombocytopenia, hyponatremia, bandemia, or transaminitis) alone and in combination with each other. RESULTS:The LR+ and LR- were statistically significant for each laboratory test individually and in combination with each other. Individual laboratory test LRs were: leukopenia, LR+ = 2.9, LR- = 0.74; thrombocytopenia, LR+ = 3.51, LR- = 0.41; bandemia, LR+ = 5.16, LR- = 0.69; hyponatremia, LR+ = 2.37, LR- = 0.46; and transaminitis, LR+ = 2.13, LR- = 0.64. Laboratory test combination LR+s were at least 2 laboratory abnormalities LR+ = 2.93, at least 3 laboratory abnormalities LR+ = 6.38, at least 4 laboratory abnormalities LR+ = 12.96, and all 5 laboratory abnormalities LR+ = 39.7. When no laboratory tests were positive, the LR- was 0.16. CONCLUSION:Laboratory abnormalities (leukopenia, thrombocytopenia, bandemia, hyponatremia, and transaminitis) alone were helpful in ruling in disease in patients with suspected disease, with greater than four or more laboratory values abnormal strongly suggestive for HGA. All five laboratory tests being normal strongly helped rule out disease.
BACKGROUND:Monkeypox (mpox) is a reemerging viral zoonosis caused by the mpox virus (MPXV) that has significant global public health implications following the 2022 and 2024 outbreaks. This review examines the evolving landscape of MPXV, focusing on its genomic organization, epidemiological shifts, phylogenetic relationship with variola virus, zoonotic transmission cycles, and the critical interventions for therapeutic targets and vaccine development. METHODS:A comprehensive narrative review of literature from 2020 to 2026 was conducted. Key thematic insights were synthesized using Bibliometrix, Scopus Analytics, and VOSviewer to analyze global trends in transmission dynamics, clinical manifestations, and therapeutic interventions. RESULTS:The review highlights the divergence of MPXV into Clades I and II, noting that while Clade IIb drove the 2022 global outbreak, the more virulent Clade Ib has dominated the 2024 reemergence. Zoonotic spillover remains a primary risk through contact with reservoirs such as rope squirrels and Gambian pouch rats, although human-to-human transmission has evolved to include rapid community spread among specific high-risk populations. Genomic analyses identify key mutations in proteins like H3L that facilitate host-specific adaptability and immune evasion. Current clinical management relies on supportive care, with targeted use of the JYNNEOS vaccine and antivirals like tecovirimat for severe cases. CONCLUSIONS:The continued reemergence of mpox underscores the necessity for an integrated "One Health" approach to surveillance and pandemic preparedness. Future research must prioritize environmental wildlife monitoring, genomic surveillance of emerging clades, and the development of multi-epitope vaccines to effectively mitigate both zoonotic spillover and human transmission.
The escalating threat of viral pandemics, dramatically illustrated by the COVID-19 crisis, has exposed the critical shortcomings of conventional reactive virology in addressing rapidly evolving pathogens. This review introduces predictive virology (PV) as an artificial intelligence (AI)-driven discipline within broader epidemic intelligence and public health surveillance that uses advanced computational tools to forecast viral threats and accelerate countermeasure design. The current review systematically examines how AI-driven approaches (e.g., machine learning and deep learning) are reshaping virology by integrating vast genomic datasets, multimodal surveillance signals, and advanced computational models to anticipate viral emergence and evolution before widespread transmission occurs. Core pillars of PV discussed include zero-shot mutational fitness and antigenic escape prediction using large protein language models; multimodal early-warning systems that fuse wastewater monitoring, digital epidemiology, mobility data, and social media; neural differential equation-based transmission modeling; generative AI for de novo design of broad-spectrum antivirals and vaccines; and ecological risk assessment of zoonotic spillovers. In retrospective benchmarks against deep mutational scanning experiments and real-world epidemiological outcomes (SARS-CoV-2 variants, influenza, and other outbreaks), several AI-powered tools have demonstrated performance comparable to or exceeding traditional methods, although prospective validation at scale remains limited. Despite remarkable progress, significant challenges persist, including data bias, overfitting to historical patterns, lack of prospective validation, and limited generalizability across settings. In addition, there are concerns about mechanistic interpretability, equitable global data integration, and responsible deployment. This review also critically addresses the ethical, governance, and equity implications of deploying predictive capabilities at a global scale. By consolidating cutting-edge AI methodologies with virological insights and acknowledging current limitations, this work provides a comprehensive framework for transitioning virology from a reactive to a truly predictive discipline, ultimately strengthening global health security and pandemic preparedness.
Mosquitoes are a source of concern because they transmit many infectious diseases, including Zika virus, chikungunya, and malaria. Accurate and rapid identification of mosquitoes is essential for disease surveillance and control. Genetic diversity in mosquito species, resulting from adaptation to different environments, leads to many differences in morphological characteristics. Traditional identification methods relying on morphology can be time-consuming and unreliable. The study aimed to identify medically and veterinary significant adult mosquito species throughout 2021 in Buraydah City, Kingdom of Saudi Arabia (KSA). We collected adult mosquitoes using Black Hole light traps in the city. The mosquitoes were morphologically identified using traditional identification keys and characterized by using the mitochondrial cytochrome oxidase c subunit I (COI) barcode regions. Although the genetic sequences we discovered do not necessarily represent new records, they do represent species with a long history of evolutionary independence, at least among adult mosquito species in KSA, especially in Buraydah city. Both morphological and molecular data identified five mosquito species: Culex pipiens (Linnaeus, 1758), Cx. sitiens (Wiedemann, 1828), Aedes aegypti (Linnaeus, 1762), Ae. caspius (Pallas, 1771), and Anopheles dthali (Patton, 1905). These species correspond to those from Kenya, India, KSA, and Iran with genetic variation. Aedes aegypti was confirmed for the first time in Buraydah city. DNA barcoding is a useful tool that can overcome inefficiencies and difficulties and complement traditional taxonomy.
BACKGROUND:Vector-borne diseases (VBDs) account for 17% of the global infectious disease burden, causing 7,00,000 deaths annually. Dengue fever remains a public health concern in India, with 32,091 cases and 32 deaths reported by June 30, 2024. Lawspet constituency in Puducherry, a dengue hotspot, provides ideal conditions for Aedes mosquito breeding. Dengue source reduction, a cost-effective strategy, targets breeding sites to interrupt the mosquito life-cycle. This study aims to assess Aedes immature positivity through a door-to-door survey in 464 households of Pedhuchettipet ward, Lawspet constituency. METHODS:A door-to-door Aedes survey was conducted in the ward Pedhuchettipet from April to May 2024. Of 960 households, 464 (48.3%) were surveyed following WHO entomological protocols. Various water-holding containers were inspected. Collected Aedes immatures were reared under controlled conditions (27°C ± 2°C, RH 70 ± 2%) until adult emergence and the adults were identified using taxonomic keys. House index (HI), Breteau index (BI), and Container index (CI) were calculated to assess dengue risk in the study area. Data were analyzed using Microsoft Excel program. RESULTS:Among 464 houses, 17 (3.6%) had Aedes immatures, with 24 positive containers. 200 immatures emerged and all identified as Aedes aegypti with a 60% emergence rate. HI (3.6%), BI (5.17%), and CI (4.36%) exceeded the dengue transmission threshold, highlighting the risk level. CONCLUSIONS:Findings indicate moderate dengue transmission risk, stressing the need for continuous surveillance and vector control. Source reduction, community interventions, and biological control are crucial in mitigating dengue risks. Environmental management and public awareness can reduce Aedes populations, lowering dengue outbreak risks in the area.
BACKGROUND:Cerebral malaria remains one of the most severe complications of Plasmodium infection and is associated with excessive oxidative stress, neuroinflammation, and high mortality. This study investigated the therapeutic and mechanistic effects of green-synthesized copper oxide nanoparticles (CuO-NPs), administered alone or in combination with quinine (QN), in mice infected with Plasmodium berghei ANKA strain. METHODS:Eighty-six BALB/c mice were randomly assigned to infected and noninfected groups. Treatments were administered orally once daily beginning 3 h postinfection for 7 consecutive days. Malaria-infected mice received QN (5 or 10 mg/kg), CuO-NPs (5 or 10 mg/kg), or combinations of CuO-NPs with low-dose QN (5 mg/kg). Survival was monitored for 28 days. Parasitemia suppression, oxidative stress biomarkers, antioxidant gene expression, and inflammatory mediators were evaluated in brain tissue homogenates. For toxicity evaluation, healthy mice received treatments for 14 days. Statistical analyses were performed using one-way analysis of variance followed by Tukey's post hoc test. RESULTS:Green-synthesized CuO-NPs exhibited spherical morphology with an average diameter of approximately 40 nm. Combined administration of CuO-NPs with QN significantly improved survival and reduced parasitemia suppression, ranging from 93.8% to the absence of microscopically detectable parasitemia relative to infected untreated controls (p < 0.001). The combinations also significantly reduced malondialdehyde and nitric oxide levels while upregulating Glutathione peroxidase (GPx) and superoxide dismutase expression. In addition, combination therapy modestly downregulated nuclear factor-kappa B p65, toll-like receptor 4, and tumor necrosis factor-alpha Messenger ribonucleic acid (mRNA) expression. Biochemical toxicity analyses revealed no significant alterations in hepatic or renal function markers following 14 days of administration. CONCLUSION:The combination of green-synthesized CuO-NPs with low-dose QN may improve antimalarial efficacy through modulation of oxidative stress and inflammatory responses in an early-intervention murine model of cerebral malaria without inducing significant short-term biochemical toxicity. However, additional mechanistic, pharmacokinetic, histopathological, and long-term safety studies are required before clinical translation can be considered.
Background: Cockroaches, including the German cockroach (Blattella germanica) can harbor and disseminate enteric human pathogens, such as Salmonella spp. Detecting Salmonella in field-collected cockroach samples is important for source attribution and monitoring public health risk, but is complicated by the low abundance of the pathogen relative to the rich and complex cockroach microbiota. Studies using different culture methods to detect Salmonella in cockroaches have reported disparate results, emphasizing the need for sensitive, accurate strategies to recover and identify Salmonella from infected cockroaches.Materials and Methods: Here, we evaluated the efficacy of several Salmonella enrichment and selective detection strategies-Rappaport-Vassiliadis (RV) broth enrichment, Tetrathionate (TT) broth enrichment, and a sequential TT followed by RV enrichment, combined with selective plating on Brilliant Green (BG) or Xylose Lysine Tergitol 4 (XLT-4) agar. Efficacy was assessed on German cockroach samples collected 1-7 days after experimental infection with a laboratory reference strain of Salmonella. The relative abundance of Salmonella in each enrichment broth was determined by 16S rRNA gene amplicon sequencing, and colony morphologies on selective plates were visually compared.Results: Overall, RV broth, TT broth, and sequential TT-RV broth strategies were equally effective at enriching Salmonella from infected cockroach samples. However, selective plating on XLT-4 was more effective at differentiating Salmonella from commensal Pseudomonas and Serratia spp. that were also enriched in RV and TT broth and produced similar colony phenotypes to Salmonella on BG agar.Conclusions: These results enhance methodological capacity for Salmonella surveillance in cockroaches and highlight the importance of considering the potential for cockroach commensals to produce false positives, interfering with accurate pathogen detection.
BACKGROUND:Ehrlichia canis is a common tick-borne pathogen in dogs and may occur without clear clinical signs, complicating diagnosis. Serological methods such as indirect immunofluorescence antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) are therefore widely used. This study aimed to determine the seropositivity of E. canis in dogs from the Kars region of Türkiye and to evaluate associated demographic, clinical, and hematological findings. MATERIALS AND METHODS:Blood samples were collected from 188 dogs of different ages, breeds, and clinical backgrounds presented to the Animal Health Education, Application, and Research Hospital of Kafkas University (Türkiye) for routine care or clinical complaints. RESULTS:E. canis seropositivity was detected at 22.34% (42/188) by IFAT and 10.64% (20/188) by ELISA. Taking IFAT as the reference, ELISA showed 50% sensitivity, 100% specificity, and 87.14% diagnostic accuracy, with moderate agreement between tests (κ = 0.598). No statistically significant differences were found in hematological parameters between seropositive and non-dual-seropositive dogs (p values ranged from 0.065 to 0.343). Among demographic and clinical variables, only age showed a significant association (p = 0.008), with higher seropositivity in older dogs; however, this should be interpreted cautiously due to small subgroup sizes. CONCLUSION:The findings indicate notable exposure to E. canis in a hospital-based sample of dogs from the Kars region, even in the absence of clinical or hematological abnormalities. Serological testing alone reflects exposure rather than active infection. Using IFAT and ELISA together may provide a more cautious approach in identifying seroreactive animals, though not necessarily improving definitive diagnostic accuracy.
BACKGROUND:Vector-borne diseases (VBDs) continue to pose major public health challenges, particularly in tropical and subtropical regions. Conventional vector control strategies, including insecticide spraying, insecticide-treated bed nets, and larvicidal interventions, have substantially reduced disease burden; however, the emergence of insecticide resistance has compromised the long-term effectiveness of these approaches. Consequently, environmentally sustainable biological and genetic vector control methods are increasingly being explored as alternative strategies. MATERIALS AND METHODS:This review comprehensively evaluates recent advances in biological and genetic approaches for mosquito vector control. The article synthesizes evidence from published laboratory, semi-field, and field studies on Wolbachia-mediated pathogen blocking, paratransgenesis using Asaia and Serratia, entomopathogenic fungi, and genetic control tools, including the sterile insect technique (SIT), incompatible insect technique (IIT), release of insects carrying a dominant lethal (RIDL), and CRISPR-Cas9-based gene drive systems. RESULTS:Multiple biological and genetic interventions demonstrated substantial reductions in mosquito populations and pathogen transmission under experimental and field conditions. Wolbachia-based systems, SIT, IIT, and RIDL approaches showed promising operational feasibility and field validation, whereas gene drive technologies exhibited strong transformative potential but remain largely experimental. However, significant challenges persist, including scalability, infrastructure requirements, ecological uncertainty, biosafety concerns, regulatory complexity, and ethical considerations. Comparative evaluation indicates that these approaches differ considerably in technological maturity, deployment readiness, and sustainability. CONCLUSIONS:Emerging biological and genetic vector control strategies represent promising alternatives to conventional insecticide-based approaches for sustainable VBD management. Rather than relying on a single universal intervention, context-specific integration of complementary approaches within existing integrated vector management frameworks, supported by community participation and effective governance, is likely to provide more durable and environmentally sustainable solutions for controlling vector-borne diseases.
BACKGROUND:Japanese encephalitis virus (JEV) is a reemerging vector-borne orthoflavivirus of significant public and animal health concern. Infection with JEV can cause inapparent, mild, or fatal encephalitis in humans and animals. The virus, which circulates primarily in Southeast Asia, has caused outbreaks in humans and livestock in nonendemic regions, leading to human disease and significant economic loss. The 2022 Australian outbreak of JEV led to renewed interest in JEV countermeasures available in the United States for veterinary use as the Australia outbreak primarily affected commercial swine farms. There are currently no antivirals or veterinary vaccines available to protect animals from JEV infection, and only one vaccine is approved for human use in the United States. In the event of a JEV introduction to the United States, vaccination of susceptible reservoirs, like domestic swine, may help prevent economic loss and JEV from establishing endemicity in the United States. METHODS:This review assesses recent advances in JEV vaccine development with an interest in veterinary applications. This systematic review was conducted by searching databases (PubMed, Web of Science, Scopus, and EBSCOhost). RESULTS:Eighty-seven research articles focused on the evaluation of novel JEV vaccine candidates were included in this review. CONCLUSION:Although there are a variety of different methodologies being employed in the development of JEV vaccine candidates, including recombinant live virus vaccines utilizing insect-only flaviviruses as vectors for JEV antigens and virus-like particle vaccines using various JEV genes to elicit protection among different JEV genotypes. While there are many promising candidates being developed, most are likely several years away from commercial production and would not be available if JEV were introduced to the United States within the next several years. Therefore, other disease control strategies should be investigated.
BACKGROUND:Rhipicephalus microplus, a one-host tick species, serves as a principal vector of tick-borne diseases in agricultural ecosystems worldwide by harboring and transmitting various pathogens through blood-feeding. In China, the climatically suitable range of R. microplus has been gradually expanding. However, the climatic suitability of R. microplus under future climate change scenarios remains unclear. METHODS:This study evaluates both current and projected climatic suitability of R. microplus by integrating climatic variables, thereby providing insight into shifts in climatic suitability under present and future climate conditions. The MaxEnt model was applied using 78 occurrence records of R. microplus collected from 1970 to 2023, along with 19 environmental variables obtained from WorldClim. By identifying the most influential environmental factors affecting the climatic suitability of R. microplus, we predicted future changes under three Shared Socioeconomic Pathways (SSP126, SSP245, SSP585) for three future periods (2021-2040, 2041-2060, and 2061-2080). RESULTS:This study indicates that the current climatically suitable areas for R. microplus are mainly located in southern China, covering approximately 1,051,406 km2, which accounts for about 10.91% of China's total land area. The minimum temperature of the coldest month (Bio06, 69.6%) and precipitation of the warmest quarter (Bio18, 20.1%) were identified as the most influential climatic variables. Under future climate scenarios, the suitable habitat for R. microplus is projected to expand and shift northward. By 2061-2080 under the SSP585, the suitable area could reach up to 2,994,700 km2, representing a 2.85-fold increase relative to its current extent. CONCLUSIONS:This study projects a significant northward expansion of climatic suitability for R. microplus in mainland China under future climate scenarios, driven primarily by rising minimum winter temperatures. These findings highlight an urgent need for proactive, climate-integrated surveillance and adaptive control strategies to mitigate the growing threat of this tick vector and its associated diseases in newly vulnerable regions.
