Experimental parasitology最新文献

{"title":"Survival and growth of M. perstans larvae in a human colon carcinoma cell line-based in vitro culture","authors":"","doi":"10.1016/j.exppara.2024.108822","DOIUrl":"10.1016/j.exppara.2024.108822","url":null,"abstract":"<div><p><em>Mansonella perstans</em> infections are widespread in Sub-Saharan Africa and Central and South America and thus can be considered as the most prevalent parasite of man in tropical Africa. In contrast to the high prevalence, knowledge about the biology of this filarial nematode is restricted and no effective treatment regimens of this ivermectin-resistant parasite is lacking. An obstacle for the research is that <em>M. perstans</em> resides in body cavities and thus have been only rarely recovered during surgery or autopsy. Therefore, alternative methods like <em>in vitro</em> culture systems need to be implemented to decipher the nature of mansonellosis and effective drugs. Previously, we have established a monkey kidney epithelial cell-based <em>in vitro</em> culture for the maintenance of <em>M. perstans</em> infective larvae (L3) up to 77 days. However, no alternative for this culture system have been postulated to allow longer survival rates and development of adult worms <em>in vitro</em>. Thus, we aim to establish an alternative <em>in vitro</em> culture system for <em>M. perstans</em> L3. <em>M. perstans</em> L3 were isolated from engorged and laboratory reared <em>Culicoides</em> midges. The larvae were then cultured in Dulbecco's Modified Eagle Medium supplemented with either 10% foetal bovine serum (FBS), 10% newborn calf serum (NCS) or 1% bovine serum albumin (BSA) together with human colon carcinoma cells (HCT-8) as feeder cells. Survival and growth were recorded. We obtained that the 10% NCS culture condition was superior allowing long-term maintenance of <em>M. perstans</em> L3 for up to 100 days and boosted growth of the parasites for up to 5-folds compared to the initial size at culture inception. Although no moulting of the L3 into L4 or adult worms could be overserved, the human colon carcinoma cell-based <em>in vitro</em> culture provides an alternative platform to analyse <em>M. perstans</em> biology and screen for novel drugs against <em>M. perstans</em>.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0014489424001255/pdfft?md5=dd9d2bd8abbf397458a5a20fe668d4dc&pid=1-s2.0-S0014489424001255-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141912359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Histological changes of oocytes of the cattle tick Rhipicephalus microplus (Canestrini, 1888) treated with copper solutions","authors":"","doi":"10.1016/j.exppara.2024.108812","DOIUrl":"10.1016/j.exppara.2024.108812","url":null,"abstract":"<div><p>Infections caused by the ectoparasite <em>Rhipicephalus microplus</em> can cause major health problems in cattle, including death. Tick control is regularly made using a range of acaricide products. As a consequence, tick populations have been heavily selected for drug resistance. The objective of this work was to determine the <em>in vitro</em> efficacy of copper chloride and sulfate (CuCl₂ and CuSO₄) solutions against <em>R</em>. <em>microplus</em>. The adult immersion test (AIT), which measures the egg-laying and egg-hatch effects, was used for the Cu-II solutions at 30, 60, 120, 240, 480, and 1000 mM, in triplicates. Distilled water and the combination of cypermethrin 20% and chlorpyrifos 50% were used as controls. Histological sections were performed from the ovaries of adult engorged female ticks treated with 240, 480, and 1000 mM of CuCl₂ and CuSO₄. We have established a histological index of the damage caused by the solutions to the tick oocytes. The overall efficacy (egg laying &amp; egg hatch) for CuCl₂ and CuSO₄ was 81.3, 82.5, 89.8, 84.5, 100.0, and 100%, and 61.7, 43.4, 62.5, 93.1, 100.0, and 98.5% respectively. Smaller oocytes were found in the Cu-II groups compared to the negative control. The histological data showed a concentration-dependent degenerative lesion of oocytes, described as cytoplasmic vacuolation and nuclear disorganization. The combination of cypermethrin and chlorpyriphos showed 100% efficacy. Cu-II solutions showed <em>in vitro</em> efficacy against adult engorged ticks being particularly harmful to oocytes. Thus, bioactive metals could be a complementary biofriendly treatment to control <em>R</em>. <em>microplus</em> and these injuries could be responsible for preventing egg hatch, and reducing pasture contamination. Safety studies are underway demonstrating the Cu-II potential in naturally infected cattle and their persistence in the environment.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141912358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A novel and low-cost cross-priming amplification assay for rapid detection of Babesia duncani infection","authors":"","doi":"10.1016/j.exppara.2024.108813","DOIUrl":"10.1016/j.exppara.2024.108813","url":null,"abstract":"<div><p><em>Babesia duncani</em>, responsible for human babesiosis, is one of the most important tick-borne intraerythrocytic pathogens. Traditionally, babesiosis is definitively diagnosed by detecting parasite DNA in blood samples and examining <em>Babesia</em> parasites in Giemsa-stained peripheral blood smears. Although these techniques are valuable for determining <em>Babesia duncani</em>, they are often time-consuming and laborious. Therefore, developing rapid and reliable <em>B</em>. <em>duncani</em> identification assays is essential for subsequent epidemiological investigations and prevention and control. In this study, a cross-priming amplification (CPA) assay was developed, combined with a vertical flow visualization strip, to rapidly and accurately detect <em>B. duncani</em> infection. The detection limit of this method was as low as 0.98 pg/μl of genomic DNA from <em>B. duncani</em> merozoites per reaction at 59 °C for 60 min. There were no cross-reactions between <em>B. duncani</em> and other piroplasms infective to humans and mammals. A total of 592 blood samples from patients bitten by ticks and experimental infected hamsters were accurately assessed using CPA assay. The average cost of the CPA assay is as low as approximately $ 0.2 per person. These findings indicate that the CPA assay may therefore be a rapid screening tool for detection <em>B. duncani</em> infection, based on its accuracy, speed, and cost-effectiveness, particularly in resource-limited regions with a high prevalence of human babesiosis.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141906300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PLGA-PEG-COOH nanoparticles are efficient systems for delivery of mefloquine to Echinococcus multilocularis metacestodes","authors":"","doi":"10.1016/j.exppara.2024.108811","DOIUrl":"10.1016/j.exppara.2024.108811","url":null,"abstract":"<div><p>Alveolar echinococcosis (AE) is a severe disease caused by the infection with the larval stage of <em>Echinococcus multilocularis</em>, the metacestode. As there is no actual curative drug therapy, recommendations to manage AE patients are based on radical surgery and prophylactic administration of albendazole or mebendazole during 2 years to prevent relapses. There is an urgent need for new therapeutic strategies for the management of AE, as the drugs in use are only parasitostatic, and can induce toxicity. This study aimed at developing a drug delivery system for mefloquine, an antiparasitic compound which is highly active against <em>E. multilocularis in vitro</em> and in experimentally infected mice. We formulated mefloquine-loaded PLGA-PEG-COOH (poly-(lactic-co-glycolic acid)) nanoparticles that exhibit stable physical properties and mefloquine content. These nanoparticles crossed the outer acellular laminated layer of metacestodes <em>in vitro</em> and delivered their content to the inner germinal layer within less than 5 min. The <em>in vitro</em> anti-echinococcal activity of mefloquine was not altered during the formulation process. However, toxicity against hepatocytes was not reduced when compared to free mefloquine. Altogether, this study shows that mefloquine-loaded PLGA-PEG-COOH nanoparticles are promising candidates for drug delivery during AE treatment. However, strategies for direct parasite-specific targeting of these particles should be developed.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0014489424001140/pdfft?md5=704dd5e7149983793351480f457d51f7&pid=1-s2.0-S0014489424001140-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141901413","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Relationship between the serum level, polymorphism and gene expression of IL-33 in samples of recurrent miscarriage Iraqi women infected with toxoplasmosis","authors":"","doi":"10.1016/j.exppara.2024.108799","DOIUrl":"10.1016/j.exppara.2024.108799","url":null,"abstract":"<div><p>One of the many warm-blooded hosts that toxoplasmosis-causing intracellular protozoan parasite <em>Toxoplasma gondii</em> can infect is humans. Cytokines are crucial to stimulate an effective immune response against <em>T. gondii</em>. Interleukin-33 (IL-33) is a unique anti-inflammatory cytokine that suppresses the immune response. The levels of cytokine gene expression are regulated by genetics, and the genetic polymorphisms of these cytokines play a functional role in this process. Single nucleotide polymorphisms (SNPs) are prognostic indicators of illnesses. This study aimed to determine whether toxoplasmosis interacts with serum levels of IL-33 and its SNP in miscarriage women as well as whether serum levels and IL-33 gene expression are related in toxoplasmosis-positive miscarriage women. Two hundred blood samples from patients and controls were collected from AL-Alawiya Maternity Teaching Hospital and AL-Yarmouk Teaching Hospital in Baghdad, Iraq from 2021 to 2022 in order to evaluate the serum level of IL-33 using ELISA test. For the SNP of IL-33, the allelic high-resolution approach was utilized, and real time-PCR was performed to assess gene expression. The results showed that compared to healthy and pregnant women, recurrent miscarriage with toxoplasmosis and recurrent miscarriage women had lower IL-33 concentrations. Additionally, there were significant differences among healthy women, pregnant women, and women with repeated miscarriage who experienced toxoplasmosis. Furthermore, no differences between patients and controls were revealed by gene expression data. The results revealed that recurrent miscarriage, pregnancy, and healthy women all had a slightly higher amount of the IL-33 gene fold. Additionally, the SNP of IL-33 data demonstrated that there was no significant genetic relationship between patients and controls. Recurrent miscarriage women with toxoplasmosis have showed significant differences from pregnant women in the genotypes GG and AA as well as the alleles A and G. There were notable variations between recurrent miscarriage with and without toxoplasmosis in terms of the genotypes AA and AC. The genotypes GG, AA, and allele A in recurrent miscarriage women with toxoplasmosis and recurrent miscarriage women is a protective factor. Taking together, there was a statistically significant negative correlation between toxoplasmosis and IL-33 gene expression, which calls for more quantitative investigation in order to fully comprehend the interaction of mRNA and protein.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141709490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Testosterone leads to Trypanosoma cruzi glycoprotein synthesis and increased of inflammatory mediators in bone marrow-derived macrophages","authors":"","doi":"10.1016/j.exppara.2024.108798","DOIUrl":"10.1016/j.exppara.2024.108798","url":null,"abstract":"<div><p>Despite all the scientific progress in recent decades to unravel the immune processes and the way the parasite bypasses the immune system, Chagas disease is still a major public health problem, affecting an estimated 3.5 million people. Among the components that may participate in the response against the parasite, testosterone has been gaining more and more visibility. Studies indicate that the parasite itself seems to carry out steroidogenesis, in which, in co-culture with androgen precursors, <em>T. cruzi</em> has been shown to produce TS, but the purpose of the TS synthesized by the parasite and how this can influence its invasion glycoproteins is still unclear unknown. The aim of this study was to evaluate the influence of testosterone in <em>Trypanosoma cruzi</em> infection on the immune response of bone marrow-derived macrophages. Bone marrow from male rats was extracted and cultured with RMPI medium containing 30% L929 cell supernatant for macrophage differentiation. The cells were incubated for 10 days and, after this period, they were seeded in 96 wells in the amount of 1 x 10⁵ cells per well. TS was added at different concentrations of 20 μM, 10 μM, 5 μM and 1 μM and then infected with the Y strain of <em>T. cruzi</em>, at a rate of 10 parasites per cell, with the culture remaining for six, 12 and 24 h. The supernatant was collected and the production of nitric oxide (NO), tumor necrosis factor (TNF) and the number of cell parasites was assessed by staining with 4′-6′-diamino-2-phenylindole (DAPI) and ranked by high Content Screening (HSC). The parasite was then cultured with the addition of TS, at the mentioned concentrations, leaving it for six and 12 h and then performing the RT-PCR of the mucins. DAPI staining revealed a significant increase in the number of parasites in cells containing TS. The exception was observed when 1 μM of hormone/well was used. A reduction in TNF production was found with 20 and 10 μM of TS for 6 h stimulation, although increased levels were observed with 5 and 1 μM, similar to the infected control. However, there was an increase in TNF production and not after 12 h. The relative expression of parasite glycoprotein 82 was increased with the presence of TS in the medium, regardless of time. Our data suggest that TS may contribute to cellular immunosuppression, increasing parasite infection in the cell, as well as inflammatory mediators that lead to cell and tissue damage in infected individuals, as well as the possible use of TS to allow their invasion into the cell hosts.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141633101","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mevalonate kinase of Leishmania donovani promotes its survival and plays a pivotal role in pathogenesis","authors":"","doi":"10.1016/j.exppara.2024.108800","DOIUrl":"10.1016/j.exppara.2024.108800","url":null,"abstract":"<div><p>The infectivity of <em>Leishmania</em> is determined by its ability to invade and evade host and its thriving capacity within the macrophage. Our study revealed the role of <em>Leishmania donovani</em> mevalonate kinase (MVK)<em>,</em> an enzyme of mevalonate pathway in visceral leishmaniasis pathogenesis. Peritoneal exudate cells (PEC)-derived macrophages from BALB/c mice were infected with wild type (WT), MVK over expressing (MVK OE) and knockdown (KD) parasites and MVK OE parasites were found to be more infective than WT and MVK KD parasites. Incubation of macrophages with MVK OE parasites declined inducible nitric oxide synthase (iNOS) expression as well as nitric oxide (NO) production, both by 2 times in comparison to WT parasites. Moreover, ∼3 fold increase in Arginase1 expression indicated that MVK might induce polarization of macrophage towards M2, favouring the survival of parasite within the macrophages. Post 24 h infection of the macrophages with mutant strains, the levels of different cytokines (TNF-α, IL-12, IL-10 and IFN-γ) were measured. Infection of macrophages with MVK OE parasites showed an increase in the level of anti-inflammatory cytokine: IL-10 while infection with MVK KD parasites exhibited an increase in the level of pro-inflammatory cytokines: TNF-α, IL-12, and IFN-γ. Hence, <em>Leishmania donovani</em> mevalonate kinase (LdMVK) modulates macrophage functions and has a significant role in pathogenesis.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141751469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The activities of suaveolol and other compounds from Hyptis suaveolens and Momordica charantia against the aetiological agents of African trypanosomiasis, leishmaniasis and malaria","authors":"","doi":"10.1016/j.exppara.2024.108807","DOIUrl":"10.1016/j.exppara.2024.108807","url":null,"abstract":"<div><p>African trypanosomiasis and malaria are among the most severe health challenges to humans and livestock in Africa and new drugs are needed. Leaves of <em>Hyptis suaveolens</em> Kuntze (Lamiaceae) and <em>Momordica charantia</em> L. (Cucurbitaceae) were extracted with hexane, ethyl acetate, and then methanol, and subjected to silica gel column chromatography. Structures of six isolated compounds were elucidated through NMR and HR-EIMS spectrometry. Callistrisic acid, dehydroabietinol, suaveolic acid, suaveolol, and a mixture of suaveolol and suaveolic acid (SSA) were obtained from <em>H. suaveolens</em>, while karavilagenin D and momordicin I acetate were obtained from <em>M. charantia</em>. The isolated biomolecules were tested against trypomastigotes of <em>Trypanosoma brucei brucei</em> and <em>T. congolense</em>, and against <em>Plasmodium falciparum</em>. The most promising EC₅₀ values were obtained for the purified suaveolol fraction, at 2.71 ± 0.36 μg/mL, and SSA, exhibiting an EC₅₀ of 1.56 ± 0.17 μg/mL against <em>T. b. brucei</em> trypomastigotes. Suaveolic acid had low activity against <em>T. b. brucei</em> but displayed moderate activity against <em>T. congolense</em> trypomastigotes at 11.1 ± 0.5 μg/mL. Suaveolol and SSA were also tested against <em>T. evansi</em>, <em>T. equiperdum</em>, <em>Leishmania major</em> and <em>L. mexicana</em> but the antileishmanial activity was low. Neither of the active compounds, nor the mixture of the two, displayed any cytotoxic effect on human foreskin fibroblast (HFF) cells at even the highest concentration tested, being 200 μg/mL. We conclude that suaveolol and its mixture possessed significant and selective trypanocidal activity.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0014489424001103/pdfft?md5=d2ed79dc6c4cd79fc249ab727dec3be3&pid=1-s2.0-S0014489424001103-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141751470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Anthelmintic activity of three selected ethnobotanical plant extracts against Strongyloides venezuelensis","authors":"","doi":"10.1016/j.exppara.2024.108801","DOIUrl":"10.1016/j.exppara.2024.108801","url":null,"abstract":"<div><p>The agropastoral farmers have employed <em>Turraea vogelii</em> <em>(TVL),</em> <em>Senna podocarpa</em> <em>(SPL),</em> and <em>Jaundea pinnata</em> (JPL) leaves for treating various diseases, including intestinal parasites in livestock and the human population in Nigeria. Gastrointestinal nematodes are highly significant to livestock production and people's health, and natural products are interesting as sources of new drugs. In this study, we evaluated the effectiveness of extracts derived from these plants in treating parasitic infections using third-stage infective larvae (L3) of <em>Strongyloides venezuelensis</em>. We obtained crude extracts using n-gexane (Hex), ethyl acetate (Ea), and methanol (Met). The extracts were analyzed for their phytochemical composition, and their ability to prevent hemolysis were tested. The mean concentrations of total phenols in SPL Hex, SPL Ea, and SPL Met were 92.3 ± 0.3, 103.0 ± 0.4, and 128.2 ± 0.5 mg/100 g, respectively. Total tannin concentrations for JPL Ea, SPL Ea, SPL Hex, and TVL Hex were 60.3 ± 0.1, 89.2 ± 0.2, 80.0 ± 0.1, and 66.6 ± 0.3 mg/100 g, respectively. The mean lethal concentration (LC₅₀) at 72 h for JPL Ea 39 (26–61) μg/mL. SPL Ea was 39 (34–45) μg/mL, and TVL Hex 31 (26–36) μg/mL. The antiparasitic activities of the extracts against L3 were dose- and time-dependent. All the extracts were slightly hemolytic to the erythrocytes. In this study, the plant extract tested demonstrated significant anti-<em>S. venezuelensis</em> activity. These phytobotanical extracts could be used to create formulations for the potential treatment of helminthiasis in animals and humans.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0014489424001048/pdfft?md5=b338fb4468da31a6ea5fb5ad43185a48&pid=1-s2.0-S0014489424001048-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141619630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Whole genome amplification and sequencing of individual Dirofilaria immitis microfilariae","authors":"","doi":"10.1016/j.exppara.2024.108806","DOIUrl":"10.1016/j.exppara.2024.108806","url":null,"abstract":"<div><p><em>Dirofilaria immitis</em> is a filarial parasitic nematode of veterinary significance. With the emergence of drug-resistant isolates in the USA, it is imperative to determine the likelihood of resistance occurring in other regions of the world. One approach is to conduct population genetic studies across an extensive geographical range, and to sequence the genomes of individual worms to understand genome-wide genetic variation associated with resistance. The immature life stages of <em>D. immitis</em> found in the host blood are more accessible and less invasive to sample compared to extracting adult stages from the host heart. To assess the use of immature life stages for population genetic analyses, we have performed whole genome amplification and whole-genome sequencing on nine (<em>n = 9</em>) individual <em>D. immitis</em> microfilaria samples isolated from dog blood. On average, less than 1% of mapped reads aligned to each <em>D. immitis</em> genome (nuclear, mitochondrial, and <em>Wolbachia</em> endosymbiont). For the dog genome, an average of over 99% of mapped reads aligned to the nuclear genome and less than 1% aligned to the mitochondrial genome. The average coverage for all <em>D. immitis</em> genomes and the dog nuclear genome was less than 1, while the dog mitochondrial genome had an average coverage of 2.87. The overwhelming proportion of sequencing reads mapping to the dog host genome can be attributed to residual dog blood cells in the microfilariae samples. These results demonstrate the challenges of conducting genome-wide studies on individual immature parasite life stages, particularly in the presence of extraneous host DNA.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":null,"pages":null},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0014489424001097/pdfft?md5=0130d8464ed6bfad7e3f983c0b1b6558&pid=1-s2.0-S0014489424001097-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141619735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}