Parasites & Vectors最新文献

{"title":"Carry-over effects in Culex species along a land use gradient with differences in microclimatic conditions.","authors":"Carmen Villacañas de Castro, Johann Musculus, Esther Timmermann, Renke Lühken, Ellen Kiel, Felix Gregor Sauer","doi":"10.1186/s13071-025-06903-y","DOIUrl":"10.1186/s13071-025-06903-y","url":null,"abstract":"<p><strong>Background: </strong>Culex pipiens sensu stricto (s.s.) and Culex torrentium are the major vectors of Sindbis, Usutu and West Nile virus in Europe. Both mosquito species typically breed in small artificial water containers (e.g. flower pots or rain barrels) and are the predominant mosquitoes in urbanised areas. The larval breeding conditions (e.g. temperature) can lead to carry-over effects on the emerging adults, which can influence their fitness traits (e.g. longevity or fecundity) and, finally, the vector capacity of a mosquito population. Our study aimed to investigate how the microclimatic heterogeneity across an urban area affects juvenile development and survival, as well as wing size, wing asymmetry and adult survival under heat stress of emerging adult Cx. pipiens s.s./Cx. torrentium.</p><p><strong>Methods: </strong>Experiments were conducted in Oldenburg (Lower Saxony, Germany) between 2021 and 2022. In a semi-field study, 45 artificial breeding habitats with 30 Cx. pipiens s.s./Cx. torrentium larvae were installed along a land use gradient, from vegetation-dominated to urban areas. The wings of all emerged mosquitoes were removed to measure wing size and wing asymmetry. Additionally, we tested the survival time of field-emerged adults exposed to 31 °C in the laboratory.</p><p><strong>Results: </strong>A piecewise structural equation model (SEM) was employed to simultaneously estimate the linear regression coefficients for various predicted relationships. The findings from the bivariate results align with the theoretical model derived from the SEM analysis. As sites with higher urbanisation indices had higher mean temperatures, mosquito development also differed along this urbanisation gradient. Results indicate that mosquitoes developing in warmer sites had shorter developmental times, and the highest juvenile survival occurred between 20 and 21 °C. Higher mean temperatures lead to lower adult survival times under heat stress and smaller wing centroid sizes. Finally, we showed that individuals with larger wing centroid sizes had lower asymmetries, which in turn also increased at higher maximum temperatures in the breeding sites, possibly indicating environmental stress.</p><p><strong>Conclusions: </strong>Our findings highlight the importance of microclimatic variation across urbanised areas on the development and fitness traits of Culex pipiens s.s./Cx. torrentium mosquitoes, emphasising the need to incorporate fine-scale microclimatic data into risk assessment models.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"256"},"PeriodicalIF":3.0,"publicationDate":"2025-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12228285/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144565077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The effect of eave and window modifications on house entry behavior of Anopheles gambiae.","authors":"Jeroen Spitzen, Martin J Lankheet, Remco P M Pieters, Miracle Gadamika, Ike Phiri, Antoine Cribellier, James G Logan, Constantianus J M Koenraadt, Kamija S Phiri, Florian T Muijres, Robert S McCann","doi":"10.1186/s13071-025-06887-9","DOIUrl":"10.1186/s13071-025-06887-9","url":null,"abstract":"<p><strong>Background: </strong>Anopheles gambiae mosquitoes transmit malaria parasites to humans mostly by biting them indoors at night, entering houses predominantly through ventilation openings such as open eaves and windows. In the study reported here, we studied how flying An. gambiae approach and enter a house, and whether barriers to reduce mosquito house entry alter mosquito flight patterns.</p><p><strong>Methods: </strong>Stereoscopic high-speed videography was used to reconstruct nearly 70,000 three-dimensional tracks of mosquitoes flying around a house during 30 experimental nights, with five combinations of closed or screened eaves and windows (eaves open - windows open; eaves open - windows closed; eaves open - windows screened; eaves closed - windows screened; eaves screened - windows screened).</p><p><strong>Results: </strong>In this study the eave and window treatments did not affect the number of mosquitoes attracted to the house. In all cases, mosquitoes were most active during the early evening, with lower but sustained activity throughout the night. Most An. gambiae mosquitoes approached the house by flying directly towards the eave in an upward sloping path with minimal left-right deviations, and most flight activity near the house was directly in front of the eave. Due to the highly attractive nature of the eave area of the house, window treatments had limited to no effect on the number of house entries when eaves were left open, highlighting the importance of closing or screening eaves to prevent mosquito house entry. For the screened eave treatment, An. gambiae spent about 10-fold more time near the eave over the course of the night compared to the time spent near the eave in the open or closed eave treatments. Moreover, these mosquitoes returned multiple times, persistently trying to enter the house. When the eaves were fully closed, mosquitoes ultimately diverted from the eave area towards the screened window, but the initial approach flights remained towards the closed eave.</p><p><strong>Conclusions: </strong>Taken together, these results demonstrate the tendency of An. gambiae to direct house entry toward the eaves, and to only divert to other house entry points as a secondary option. The persistent mosquito flight near screened eaves may provide guidance for the placement of outdoor vector control tools.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"251"},"PeriodicalIF":3.5,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12231866/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144560736","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CRISPR technologies for the control and study of malaria-transmitting anopheline mosquitoes.","authors":"Andrea L Smidler, Omar S Akbari","doi":"10.1186/s13071-025-06905-w","DOIUrl":"10.1186/s13071-025-06905-w","url":null,"abstract":"<p><p>Malaria is one of the deadliest diseases on the planet, killing approximately 600,000 people annually, and is transmitted by the bite of an anopheline mosquito. Anophelines, and the diseases they transmit, have changed the course of history and the fate of nations, and their successful control promises to end the transmission of malaria. With the advent of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technologies, the study and control of these deadly pests have been revolutionized. As the release of genetically modified anophelines is being considered, here we outline the advances in CRISPR/Cas9 technologies and how they have revolutionized the study of anopheline basic biology and the development of innovative vector control strategies. We outline the major findings of CRISPR-based basic biological research into traits relevant for vector control including, but not limited to, olfaction, chemosensation, neurobiology, and reproduction. Further, we summarize the advancements in CRISPR-based innovative vector control strategies, such as the precision-guided sterile insect technique (pgSIT), inherited female elimination by genetically encoded nucleases to interrupt alleles (IFEGENIA), X-shredder, Y-linked editors, and gene drives. All in all, this review summarizes the basic biological and vector control research undertaken using CRISPR since its advent approximately a decade ago.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"252"},"PeriodicalIF":3.0,"publicationDate":"2025-07-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12226854/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144560735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pentatrichomonas hominis induces extracellular traps formation of macrophages via the TLR2/NADPH/PAD4 pathway.","authors":"Qi-An Zhai, Xi-Chen Zhang, Hong-Bo Zhang, Jian-Hua Li, Peng-Tao Gong, Xiao-Cen Wang, Xin Li, Xu Zhang, Nan Zhang","doi":"10.1186/s13071-025-06840-w","DOIUrl":"10.1186/s13071-025-06840-w","url":null,"abstract":"<p><strong>Background: </strong>Pentatrichomonas hominis (P. hominis) is a newly identified pathogenic zoonotic protozoan belonging to the Trichomonadidae family. P. hominis mainly parasitizes the cecum and colon of humans and other mammals, and it can cause diarrhea. Recently, macrophage extracellular traps (METs) have been shown to play an important role in resistance to parasitic infections. However, it remains unclear whether the release of METs by macrophages contributes to P. hominis resistance, and the underlying mechanism of this association has yet to be elucidated.</p><p><strong>Methods: </strong>Scanning electron microscopy (SEM) and immunofluorescence staining were used to determine whether P. hominis induced the formation of METs in mouse peritoneal macrophages to capture and immobilize the parasite as well as the components of METs, including the DNA backbone, myeloperoxidase (MPO), and histone H3. Reactive oxygen species (ROS) and signaling pathway inhibitor assays revealed that the mechanism of P. hominis-induced MET formation was dependent on nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase activation, store-operated calcium entry (SOCE),, and peptidyl arginine deiminase 4 (PAD4) activation. The toll-like receptors 2 (TLR2), extracellular regulated protein kinase 1/2 (ERK1/2) and p38 MAPK signaling pathway were also involved in this process. Trypan blue staining revealed a gradual decrease in the survival rate of P. hominis as the coculture time increased. Trypan blue staining also revealed an increase in the proportion of macrophages.</p><p><strong>Results: </strong>The results of this study indicate that P. hominis can induce the release of METs via the TLR2/NADPH/PAD4 pathways and that METs have a trapping and killing effect on P. hominis.</p><p><strong>Conclusions: </strong>This was the first study to reveal that PAD4 and TLR2 were found to be involved in the development of parasite-induced METs, thus providing guidance for further research on the mechanisms of host innate immunity against parasitic infection.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"248"},"PeriodicalIF":3.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12220668/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144541840","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"SNP-based molecular diagnostic platform: rapid single-step identification of Theileria annulata and its buparvaquone-resistant strains.","authors":"Jin Che, Yijun Chai, Shuaiyang Zhao, Jinming Wang, Jianxun Luo, Guiquan Guan, Hong Yin, Wei Li","doi":"10.1186/s13071-025-06884-y","DOIUrl":"10.1186/s13071-025-06884-y","url":null,"abstract":"<p><strong>Background: </strong>Theileria annulata, a tick-borne protozoan that causes tropical theileriosis, poses a serious threat to livestock production in endemic regions. The emergence of resistance to buparvaquone, the primary chemotherapeutic treatment, has been attributed to acquired mutations in the cytochrome b (Cytb) gene, with identical resistance-associated polymorphisms observed in both laboratory-adapted strains and field isolates from China.</p><p><strong>Methods: </strong>A dual probe-specific real-time polymerase chain reaction (PCR) assay was developed to detect point mutations in the Cytb gene. The specificity, sensitivity, and reproducibility of the assay were validated, and its field applicability was evaluated via cattle blood samples (n = 531) collected from five endemic Chinese provinces.</p><p><strong>Results: </strong>Six point mutations were identified in the Cytb gene, and the developed dual probe-specific real-time PCR assay simultaneously detected T. annulata infection and distinguished between the buparvaquone-sensitive and buparvaquone-resistant genotypes. The assay demonstrated a detection limit of 1 × 10¹ copies/μl, high specificity, and satisfactory repeatability, with results consistent with those of Sanger sequencing. Field screening revealed a 21.7% (115/531) prevalence of T. annulata and a 4.3% (23/531) occurrence of resistant genotypes. Moreover, two-dimensional scatterplot visualization enabled clear genotype discrimination without post-PCR processing.</p><p><strong>Conclusions: </strong>The developed dual probe-specific real-time PCR assay enables efficient detection of buparvaquone-resistant genotypes, providing important implications for guiding the treatment of tropical theileriosis and enhancing epidemiological surveillance of emerging resistance in endemic regions.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"247"},"PeriodicalIF":3.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12219782/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144541804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A sialic acid-binding protein in Toxoplasma gondii contains a conserved globular domain in apicomplexan parasites.","authors":"Xiaoyu Sang, Yize Liu, Yiwei Zhang, Ran Chen, Ying Feng, Ning Jiang, Qijun Chen","doi":"10.1186/s13071-025-06845-5","DOIUrl":"10.1186/s13071-025-06845-5","url":null,"abstract":"<p><strong>Background: </strong>Apicomplexan protozoans employ an intricate invasion mechanism involving dynamic interactions with host cells, characterized by sequential secretion of adhesins and lectins. Our laboratory previously identified TgSABP1, a novel Toxoplasma gondii adhesin, demonstrating specific binding affinity for sialic acid (SA) receptors on host cell surfaces. However, the structural determinants governing SA recognition by this adhesin remain undefined.</p><p><strong>Methods: </strong>Three-dimensional structural predictions of TgSABP1 and homologous proteins were generated using AlphaFold2. Bio-layer interferometry (BLI) quantified the binding affinities between the recombinant proteins and ligands. Competitive BLI assays evaluated small molecules that potentially inhibit the TgSABP1-sialyllactose interactions. Molecular docking simulations employing AutoDock Vina software elucidated ligand-binding site interactions. In vitro invasion inhibition assays were performed to assess the therapeutic potential of lead compounds targeting TgSABP1 against T. gondii tachyzoites.</p><p><strong>Results: </strong>AlphaFold2 structural predictions revealed that TgSABP1 and its homologues contain a conserved globular domain (pLDDT > 90) with significant structural homology (with root-mean-square deviation [RMSD] < 4 Å) to a Plasmodium falciparum invasion-related protein PfIMP2 (PDB: 5LG9). BLI quantification demonstrated the micromolar binding affinities of the recombinant proteins for 3'-sialyllactose-polyacrylamide (PAA) and 6'-sialyllactose (6'SL)-PAA. Intriguingly, although recombinant TgSABP1 showed stronger lactose binding (K_D = 0.02 ± 0.01 M) compared to SA (K_D = 2.07 ± 0.45 M), only the latter exhibited an inhibition on the TgSABP1-6'SL-PAA interaction. Virtual screening of Food and Drug Administration (FDA)-approved compounds identified eltrombopag as a high-affinity molecule (ΔG_bind = -8.3 kcal/mol) targeting the SA-binding pocket in TgSABP1. Functional validation demonstrated that eltrombopag effectively blocked the TgSABP1/6'SL-PAA interaction and significantly decreased host cell invasion of T. gondii tachyzoites.</p><p><strong>Conclusions: </strong>Our study reveals a conserved globular domain of apicomplexan parasites as a novel SA-binding domain. Structural and functional characterization demonstrates its critical role in mediating TgSABP1-host cell interactions. Targeting this SA-binding pocket with eltrombopag effectively decreased T. gondii tachyzoite invasion, suggesting its therapeutic potential as an anti-invasion target. These findings not only elucidate a conserved mechanism underlying host receptor recognition in apicomplexans, but also establish a structural framework for the rational design of broad-spectrum inhibitors targeting invasion-related lectin domains.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"239"},"PeriodicalIF":3.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12210739/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144541823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Simulium larvae susceptibility to temephos and the effect of 10 weeks of treatment of the Mbende tributary in the Nkam-Wouri River drainage of Cameroon on larval density and adult fly biting rates.","authors":"Raphael Awah Abong, Relindis Ekanya, Theobald Mue Nji, Fanny Fri Fombad, Glory N Amambo, Innocentia Ngong, Franck Noel Nietcho, Kebede Deribe, Benjamin Biholong, Flobert Njiokou, Same Ekobo, Charles Wondji, Peter Enyong, Samuel Wanji","doi":"10.1186/s13071-025-06837-5","DOIUrl":"10.1186/s13071-025-06837-5","url":null,"abstract":"<p><strong>Background: </strong>Despite over 18 years of annual ivermectin mass drug administration (MDA) in Cameroon's Nkam-Wouri River drainage, onchocerciasis transmission persists. Several reasons, including multiple breeding sites and abundant vector populations, contribute to ongoing transmission. High vector abundance also causes a biting nuisance to local populations. The change in paradigm from onchocerciasis control to elimination may not be achieved if alternative control measures are not used. There is a need to complement ivermectin MDA with other strategies. This study tested the susceptibility of Simulium larvae to temephos insecticide and monitored the effect of 10 weeks of ground larviciding on the larval density and black fly population.</p><p><strong>Methods: </strong>Simulium breeding sites along the course of three rivers within the Solle transmission zone in the Nkam-Wouri River drainage were identified. Seven temephos concentrations (0.001-0.1 mg/l) were tested on freshly collected Simulium larvae for susceptibility. Simulium biting rates were monitored using human landing catches before and during 10 weeks of ground larviciding. Fishing was used to assess the abundance and diversity of large aquatic fauna, while the presence and diversity of small invertebrate fauna were assessed during the collection of larvae, as they are usually found on the same substrates in the river. Ground larviciding was conducted using the spraying method at two dosing points.</p><p><strong>Results: </strong>Six breeding sites were identified. Larval mortality decreased with temephos concentration, with 100% mortality observed at 0.1-0.025 mg/l. The non-target fauna included various fish species, crabs, crayfish, and small invertebrates. Ground larviciding cleared larvae from identified substrates and reduced adult fly biting rates by 82.8% (from 900 flies/man/day at the beginning to 180 flies/man/day at the end), a statistically significant decrease (χ² = 1351.5, P < 0.001).</p><p><strong>Conclusions: </strong>Simulium larvae showed susceptibility to temephos. Clearance of larvae from traps and identified natural substrates, and a significant reduction in the Simulium biting rates were observed.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"242"},"PeriodicalIF":3.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12210698/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144541842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Widespread distribution of Aedes aegypti larvae, a potential risk of arbovirus transmission in the Grand Lomé health region, Togo, West Africa.","authors":"Kossivi I Akagankou, Koffi M Ahadji-Dabla, Daniel Romero-Alvarez, Juan-Carlos Navarro, Leonardo D Ortega-López, Manuel Villanueva-Sarmiento, Komlan G J N'Tsoukpoe, Edoh Koffi, Yovo Kondo, Adjo A Amekudi, Yawo Apetogbo, Audrey Lenhart, Guillaume K Ketoh","doi":"10.1186/s13071-025-06835-7","DOIUrl":"10.1186/s13071-025-06835-7","url":null,"abstract":"<p><strong>Background: </strong>Understanding the population dynamics and geographical range of Aedes aegypti is critically important for arbovirus vector surveillance and control. Little is known about the current distribution and seasonality of Ae. aegypti in Grand Lomé, Togo. We developed an investigation to determine whether Ae. aegypti was present across Lomé communes during a 1-year collection period.</p><p><strong>Methods: </strong>Mosquito ovitraps (n = 70) were deployed across the 13 communes in the Grand Lomé health region and were examined between May 2022 and April 2023. Generalized linear mixed models (GLMMs) were applied to investigate the relationship between larval collections and seasonality. The European Space Agency (ESA) WorldCover 10 m 2020 product was used to represent different land cover classes and to determine whether sites with higher larval numbers differed from sites with lower numbers.</p><p><strong>Results: </strong>A total of 52,768 Ae. aegypti larvae were collected across the 13 communes of Grand Lomé. The highest incidence of Ae. aegypti larvae was observed in the commune of Bè-Ouest (= 122.74 per 1000 population). Agoè-Nyivé was the commune with the lowest incidence over the entire study period. There was a statistically significant difference in Ae. aegypti larval counts between the rainy and dry seasons. Eight land-use classes were represented by the ESA 10 m product in Grand Lomé, with the built-up category being the most common. We found a significant relationship between larval abundance categories and land cover classes.</p><p><strong>Conclusions: </strong>This study shows that Ae. aegypti larvae can be found across all communes of the Grand Lomé region in both the rainy and dry seasons, especially in ovitraps surrounded by built-up land cover category. The results of this study could be useful in guiding disease vector surveillance and control efforts due to the potential imminent risk of upcoming dengue outbreaks.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"241"},"PeriodicalIF":3.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12211295/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144541806","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a novel ddPCR assay for the simultaneous detection of the protozoan parasites Leishmania infantum and Leishmania tarentolae.","authors":"Alessandro Alvaro, Giulia Maria Cattaneo, Ilaria Varotto-Boccazzi, Riccardo Molteni, Jairo Alfonso Mendoza-Roldan, Matteo Brilli, Matilde Silvia Conconi, Virginia Giovagnoli, Alessandro Manenti, Domenico Otranto, Claudio Bandi, Sara Epis","doi":"10.1186/s13071-025-06871-3","DOIUrl":"10.1186/s13071-025-06871-3","url":null,"abstract":"<p><strong>Background: </strong>Leishmaniases, caused by protozoan parasites of the genus Leishmania, are vector-borne diseases occurring mainly in the tropics and subtropics of the world, as well as in the Mediterranean Basin. In this area, the mammalian pathogen Leishmania infantum is endemic, along with the reptile-associated Leishmania tarentolae. The two species occur in sympatry, and there is evidence that the exposure to L. tarentolae in mammalian hosts may elicit a protective immune response towards pathogenic Leishmania species. Accurate detection methods for both species are therefore crucial for gathering comprehensive information on the epidemiology of leishmaniases. In microbiological diagnosis, limits in detection performance imply the risk of false negatives and other issues, which highlights the need for sensitive methods.</p><p><strong>Methods: </strong>Here, we developed a droplet digital polymerase chain reaction assay targeting the kinetoplast minicircle DNA, for the simultaneous and differential detection of L. infantum and L. tarentolae. The assay features primers designed to bind to both species and species-specific probes. The assay was validated on three cultured isolates for each species, whose cells were spiked into Leishmania-negative dog blood, and on Leishmania-positive sand flies. Sensitivity was assessed with testing serial dilutions, and specificity was evaluated by assessing the cross-reactivity of the probes with the controls of Leishmania-free dog blood and male sand fly DNA.</p><p><strong>Results: </strong>The assay demonstrated high sensitivity, with a limit of detection corresponding to one Leishmania cell in the reaction mix for isolates of both L. infantum and L. tarentolae. Limited cross-reaction of the L. tarentolae-targeting probe was observed on L. infantum isolates. No cross-reaction was observed with the controls of Leishmania-free dog blood and male sand flies.</p><p><strong>Conclusions: </strong>The protocol can represent a valuable method for comprehensive surveillance in both canine hosts and sand flies in areas in which L. infantum and L. tarentolae occur in sympatry.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"243"},"PeriodicalIF":3.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12210423/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144541838","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New findings on the male reproductive system and spermatozoa of Aedes aegypti (Diptera: Culicidae).","authors":"Henrique Barbosa da Silva, Renata Cristina Barbosa, Paulo Henrique Rezende, Dayvson Ayala-Costa, José Lino-Neto","doi":"10.1186/s13071-025-06808-w","DOIUrl":"10.1186/s13071-025-06808-w","url":null,"abstract":"<p><strong>Background: </strong>Aedes aegypti is one of the most important arbovirus vectors, characterized by its widespread distribution and exceptional reproductive capacity. This study reexamines the male reproductive system (MRS) of this species, focusing on its morphology throughout post-embryonic development and the structure of its spermatozoa.</p><p><strong>Methods: </strong>We analyzed the MRS of A. aegypti in the larval L4, pupal, and adult stages using bright-field light microscopy, fluorescent microscopy, and transmission electron microscopy techniques. Spermatozoa measurements were made using the ImageJ software.</p><p><strong>Results: </strong>In L4 larvae, the MRS was composed of the two testes, a thin deferent duct, and a pair of seminal vesicles. The MRS is fully developed in pupae and adults, with two testes, deferent ducts, seminal vesicles, accessory glands, and an ejaculatory duct. Histological sections revealed that each testis is formed by a single follicle, which appeared to spiral at all stages. In pupae and adults, the testes showed germ cells at different stages of development, while the goblet portion of the deferent duct contained cytoplasmic bodies and spermatozoa. In adults, the seminal vesicles were filled with spermatozoa soon after emergence. Secretions from accessory glands were of the apocrine type. The spermatozoa were thin and long, measuring around 335 µm in length. Ultrastructural analysis revealed a very short acrosome covering the apical nucleus, in the flagellar region, an axoneme with the 9 + 9 + '1' microtubule pattern typical of mosquitoes, and two mitochondrial derivatives along the flagellum, narrowing at the terminal portion.</p><p><strong>Conclusions: </strong>Our analysis revealed a clear link between testicular development and spermatogenesis. In addition, we identified seminal vesicles at all life stages and accessory glands visible only in pupae and adults. The characterization of sperm structure and ultrastructure indicated similarities with other mosquito species. Finally, our study provided valuable information that may support research in comparative biology and reproduction.</p>","PeriodicalId":19793,"journal":{"name":"Parasites & Vectors","volume":"18 1","pages":"246"},"PeriodicalIF":3.0,"publicationDate":"2025-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12217529/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144541839","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}