Experimental parasitology最新文献

{"title":"Anthelmintic activity of three selected ethnobotanical plant extracts against Strongyloides venezuelensis","authors":"Ibukun O. Busari ,&nbsp;Joel H. Elizondo-Luévano ,&nbsp;Olapeju O. Aiyelaagbe ,&nbsp;Kehinde O. Soetan ,&nbsp;Olaniyi J. Babayemi ,&nbsp;Oscar Gorgojo-Galindo ,&nbsp;Antonio Muro ,&nbsp;Belén Vicente ,&nbsp;Julio López-Abán","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":"263 ","pages":"Article 108801"},"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":"Rosemonde I. Power ,&nbsp;Stephen R. Doyle ,&nbsp;Jan Šlapeta","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":"263 ","pages":"Article 108806"},"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}

{"title":"Metabolic disruptions in Biomphalaria glabrata induced by Heterorhabditis bacteriophora HP88: Implications for entomopathogenic nematodes in biological control","authors":"Victor Menezes Tunholi ,&nbsp;Ludimila Santos Amaral ,&nbsp;Natânia do Carmo Sperandio ,&nbsp;Lorena Souza Castro Altoé ,&nbsp;Vinícius Menezes Tunholi-Alves ,&nbsp;Juberlan Garcia ,&nbsp;Ester Maria Mota ,&nbsp;Melissa Carvalho Machado do Couto-Chambarelli ,&nbsp;Caio Márcio de Oliveira Monteiro ,&nbsp;Isabella Vilhena Freire Martins","doi":"10.1016/j.exppara.2024.108804","DOIUrl":"10.1016/j.exppara.2024.108804","url":null,"abstract":"<div><p>Research on the use of entomopathogenic nematodes (EPNs) as a potential tool for the biological control of invertebrates has been growing in recent years, including studies involving snails with One Health importance. In this study, the effect of exposure time (24 or 48 h) of <em>Heterorhabditis bacteriophora</em> HP88 on the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as well as the concentration of total proteins, uric acid, and urea in the hemolymph of <em>Biomphalaria glabrata</em>, were investigated. The concentrations of these metabolic markers were measured weekly until the end of the third week after exposure. Along with a significant reduction in total protein levels, a significant increase (p &lt; 0.01) in uric acid and urea contents in the hemolymph of <em>B. glabrata</em> exposed to <em>H. bacteriophora</em> was observed. The accumulation of urea in these mollusks could lead to deleterious effects due to its high toxicity, inducing significant cell damage. Variations in transaminase activities were also observed, with snails exposed to EPNs showing significantly higher values (p &lt; 0.01) than individuals in the control group, both for ALT and AST. These results indicate that experimental exposure to infective juveniles of <em>H. bacteriophora</em> causes significant alterations in the metabolic pattern of <em>B. glabrata</em>, compromising the maintenance of its homeostasis. Finally, exposure for 48 h caused more damage to the planorbid in question compared to snails exposed for 24 h, suggesting that the exposure time may influence the intensity of the host's response.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"263 ","pages":"Article 108804"},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141633100","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":"Production and evaluation of a new set of recombinant antigens for the serological diagnosis of human cysticercosis","authors":"Jihen Melki ,&nbsp;Thierry-Borel N'dri Kouadio ,&nbsp;Mireille Nowakowski ,&nbsp;Zara Razafiarimanga ,&nbsp;Man-Koumba Soumahoro ,&nbsp;Stephane Peltres ,&nbsp;Ronan Jambou","doi":"10.1016/j.exppara.2024.108803","DOIUrl":"10.1016/j.exppara.2024.108803","url":null,"abstract":"<div><p>Human cysticercosis caused by <em>Taenia soliun</em> (<em>T</em>. <em>soliun</em>) is endemic in certain areas of Latin America, Asia and Sub-Saharan Africa. Neurocysticercosis (NCC) is mainly diagnosed by neuroimaging, which, in most cases, is unavailable in endemic areas. Due to their high sensitivity and specificity, serological tests such as enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) based on the <em>glycosylated</em> fraction of the cyst <em>CS50 are widely used for the detection of the anti-cysticercus IgG antibodies despite their significant cost and the need of cysticercus material.</em> Given their cost-effectivess and simplicity, immunoassays based on recombinant proteins could provide new alternatives for human cysticercosis diagnosis: such tests would be aimed at screening those people living in remote areas who need further examination. <em>To date, however, no test using recombinant antigens i</em>s commercially available.</p><p>Herein, five recombinant proteins (R14, R18, R93.1, R914.1, and R915.2) were produced, three of which (R93.1, R914.1, and R915.2) were newly identified from the cyst fluid. Evaluation of the diagnostic performance of these recombinant antigens by ELISA was done using sera from 200 epileptic and non-epileptic individuals in comparison with the WB-CS50 as the reference serological method.</p><p>Recombinant proteins-based ELISA showed a level of diagnostic performance that is inferior than the reference serological method, but similar to that of the native antigen ELISA for human cysticercosis (commonly used for screening). Further optimization of expression conditions is still needed in order to improve proteins solubility and enhance diagnostic performance for human cysticercosis detection. However, this preliminary evaluation of the recombinant antigens has shown their potential valuable use for screening cysticercosis in patients with epilepsy attending dispensaries in remote areas. Future studies should be conducted to evaluate our recombinant antigens in a large group of patients with different stages of NCC, and in correlation with imaging findings.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"263 ","pages":"Article 108803"},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0014489424001061/pdfft?md5=bc7bf98906b490f412a6443a421318ed&pid=1-s2.0-S0014489424001061-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141619734","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":"The potential prophylactic and therapeutic efficacy of progesterone and mifepristone on experimental trichinellosis with ultra-structural studies","authors":"Doaa A. Hamdy ,&nbsp;Enas Y. Abu-Sarea ,&nbsp;Hala M. Elaskary ,&nbsp;Eman Ahmed Abd Elmaogod ,&nbsp;Gehad Abd-Elftah Abd-Allah ,&nbsp;Heba Abdel-Tawab","doi":"10.1016/j.exppara.2024.108805","DOIUrl":"10.1016/j.exppara.2024.108805","url":null,"abstract":"<div><p>Right up to now, there has not been an effective or safe therapy for trichinellosis. Thus, this study aimed to determine the efficacy of prophylactic and therapeutic regimens of progesterone and mifepristone on the intestinal and muscular phases of experimental <em>Trichinella spiralis</em> infection compared to albendazole. Seven distinct groups of mice were divided as follows: negative, positive, and drug control groups, as well as prophylactic and treatment groups using mifepristone and progesterone. Mice were sacrificed on the 7th and 37th days after infection. Treatment efficacy was evaluated using parasitological techniques, histopathological examination, immunohistochemical staining, and ultrastructural morphological analysis of adult worms by scanning electron microscopy. The mice groups received progesterone (300 ng/ml) and mifepristone (100 ng/ml). They demonstrated a significant improvement in intestinal and muscular inflammation and a statistically significant decline in the adult worm burden and encysted larvae (P &lt; 0.001). Moreover, immunohistochemical staining of vascular endothelial growth factor and mucosal mast cell analyses were coincided with the obtained parasitological results. There was notable destruction and degeneration of the adult worm tegument by using both drugs. The current study pointed out that progesterone and mifepristone may provide new insights regarding the development of vaccines and drug protocols to treat trichinellosis through their combined action in reducing the inflammation, affecting the intestinal immune cell, and decreasing the adult worm burden, and larval capsule development.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"263 ","pages":"Article 108805"},"PeriodicalIF":1.4,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141733877","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":"Antiparasitic activity of chalcones analogue against Trichomonas vaginalis: biochemical, molecular and in silico aspects","authors":"Bárbara da Rocha Fonseca ,&nbsp;Raquel Nascimento das Neves ,&nbsp;Adriane Leites Strothmann ,&nbsp;Ângela Sena-Lopes ,&nbsp;Caroline Carapina da Silva ,&nbsp;Paloma Taborda Birmann ,&nbsp;Lucielli Savegnago ,&nbsp;Claudio Martin Pereira de Pereira ,&nbsp;Sibele Borsuk","doi":"10.1016/j.exppara.2024.108809","DOIUrl":"10.1016/j.exppara.2024.108809","url":null,"abstract":"<div><p><em>Trichomonas vaginalis</em> is the etiologic agent of trichomoniasis, a worldwide distributed sexually transmitted infection (STI) that affects the genitourinary tract. Even though this disease already has a treatment in the prescription of drugs of the 5-nitroimidazole class, described low treatments adhesion, adverse side effects and cases of resistant isolates demonstrate the need for new formulations. With this in mind, chalcones emerge as a potential alternative to be tested, being compounds widely distributed in nature, easy to chemically synthesize and presenting several biological activities already reported. In this experiment, we evaluated the antiparasitic activity of 10 chalcone at a concentration of 100 μM against ATCC 30236 <em>T. vaginalis</em> isolates, considering negative (live trophozoites), positive (Metronidazole 100 μM) and vehicle (DMSO 0.6%) controls. Compounds 3a, 3c, 3 g and 3i showed promising results, with MICs set at 70 μM, 80 μM, 90 μM and 90 μM, respectively (p &lt; 0,05). Cytotoxicity assays were performed on VERO and HMVII cell lines and revealed low inhibition rates at concentrations bellow 20 μM. To elucidate a possible mechanism of action for these molecules, the DPPH, ABTS and FRAP assays were performed, in which none of the four compounds presented antioxidant activity. Assays to verify ROS and lipid peroxidation in the parasite membrane were performed. None of the tested compounds identified ROS accumulation after incubation with trophozoites. 3 g molecule promoted an increase in MDA production after incubation. Results presented in this paper demonstrate the promising trichomonicidal profile, although further tests are still needed to optimize their performance and better elucidate the mechanisms of action involved.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"265 ","pages":"Article 108809"},"PeriodicalIF":1.4,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141878589","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":"Microemulsions strongly promoted the activity of α-bisabolol against different Leishmania species and its skin permeation","authors":"Quesia Nery dos Santos ,&nbsp;Daiane Caroline S. Teles ,&nbsp;Guilherme Rodolfo S. de Araujo ,&nbsp;Odeanny Vitória A. Lima ,&nbsp;Luiz André S. Silva ,&nbsp;Rita de Cássia V. de Carvalho ,&nbsp;Valéria Carlos de Sousa ,&nbsp;Saulo S. Matos ,&nbsp;Amanda Mendonça B. Costa ,&nbsp;Valter V. Andrade-Neto ,&nbsp;Eduardo Caio Torres-Santos ,&nbsp;Adriano Antunes de S. Araújo ,&nbsp;Victor Hugo V. Sarmento ,&nbsp;Fernando Aécio de Amorim Carvalho ,&nbsp;Rogéria de S. Nunes ,&nbsp;Ana Amélia M. Lira","doi":"10.1016/j.exppara.2024.108808","DOIUrl":"10.1016/j.exppara.2024.108808","url":null,"abstract":"<div><p>This study aimed to develop microemulsions (MEs) containing α-bisabolol for the topical treatment of cutaneous leishmaniasis (CL). Initially, pseudoternary phase diagrams were developed using α-bisabolol as the oil phase, Eumulgin® CO 40 as the surfactant, Polymol® HE as the co-surfactant, and distilled water as the aqueous phase. Two transparent liquid systems (TLS) containing 5% of α-bisabolol were selected and characterized (F5E25 and F5EP25). Next, skin permeation and retention assays were performed using Franz cells. The interaction of the formulation with the stratum corneum (SC) was evaluated using the FTIR technique. The cytotoxicity was evaluated in murine peritoneal macrophages. Finally, the antileishmanial activity of microemulsions was determined in promastigotes and amastigotes of <em>L. amazonensis</em> (strain MHOM/BR/77/LTB 0016). As a result, the selected formulations showed isotropy, nanometric size (below 25 nm), Newtonian behavior and pH ranging from 6.5 to 6.9. The MEs achieved a 2.5-fold increase in the flux and skin-permeated amount of α-bisabolol. ATR-FTIR results showed that microemulsions promoted fluidization and extraction of lipids and proteins of the stratum corneum, increasing the diffusion coefficient and partition coefficient of the drug in the skin. Additionally, F5E25 and F5EP25 showed higher activity against promastigotes (IC₅₀ 13.27 and 18.29, respectively) compared to unencapsulated α-bisabolol (IC₅₀ 53.8). Furthermore, F5E25 and F5EP25 also showed antileishmanial activity against intracellular amastigotes of <em>L. amazonensis,</em> with IC₅₀ 50 times lower than free α-bisabolol and high selectivity index (up to 15). Therefore, the systems obtained are favorable to topical administration, with significant antileishmanial activity against <em>L. amazonensis</em> promastigotes and amastigotes, being a promising system for future <em>in vivo</em> trials.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"265 ","pages":"Article 108808"},"PeriodicalIF":1.4,"publicationDate":"2024-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141878590","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":"Kato-Katz slide preservation technique: Extension of viability and the benefits for schistosomiasis control programs","authors":"Rodrigo Loyo ,&nbsp;Elainne Christine Souza Gomes ,&nbsp;Otavio Sarmento Pieri ,&nbsp;Emília Carolle Azevedo de Oliveira ,&nbsp;Wheverton Ricardo Correia Nascimento ,&nbsp;Constança Simões Barbosa","doi":"10.1016/j.exppara.2024.108802","DOIUrl":"10.1016/j.exppara.2024.108802","url":null,"abstract":"<div><p>The World Health Organization recommends the use of the Kato-Katz method in the procedures of schistosomiasis control programs. Studies show the importance of a fast reading of the slides due to the decline of their viability, with the appearance of fungi or desiccation of the sample, which hinders diagnosis. It is necessary to establish a procedure to improve the long-term preservation of these Kato-Katz slides in order to accomplish the following: (1) preserve the slides for future quality control procedures and readings; (2) allow for the production of durable materials for training; and (3) train health professionals involved in diagnosing schistosomiasis. Therefore, this study aims to test a slide preservation methodology for these purposes. The results showed that the modifications made to the experimental slides demonstrated that egg loss was within the expected range and the limit accepted by quality control standards, as well as improved the diagnostic durability of the slides during the preservation times tested. We concluded that the application of the preservation technique to the slides promoted stabilization and permanence for long-term storage.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"265 ","pages":"Article 108802"},"PeriodicalIF":1.4,"publicationDate":"2024-07-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141751468","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":"Phylogenetic inferences based on distinct molecular markers reveals a novel Babesia (Babesia pantanalensis nov. sp.) and a Hepatozoon americanum-related genotype in crab-eating foxes (Cerdocyon thous)","authors":"Ana Cláudia Calchi ,&nbsp;Laíza de Queiroz Viana Braga ,&nbsp;Ricardo Bassini-Silva ,&nbsp;Ana Carolina Castro-Santiago ,&nbsp;Heitor Miraglia Herrera ,&nbsp;João Fábio Soares ,&nbsp;Darci Moraes Barros-Battesti ,&nbsp;Rosangela Zacarias Machado ,&nbsp;Fabiana Lopes Rocha ,&nbsp;Marcos Rogério André","doi":"10.1016/j.exppara.2024.108786","DOIUrl":"10.1016/j.exppara.2024.108786","url":null,"abstract":"<div><p>Piroplasmids and <em>Hepatozoon</em> spp. Are apicomplexan protozoa that may cause disease in several canid species. The present study aimed to expand the knowledge on the diversity of piroplasmids and <em>Hepatozoon</em> in crab-eating foxes (<em>Cerdocyon thous</em>; n = 12) sampled in the Pantanal of Mato Grosso do Sul State, central-western Brazil. PCR assays based on the 18S rRNA were used as screening. Three (25%) and 11 (91.7%) were positive for piroplasmids and <em>Hepatozoon</em> spp., respectively. Co-infection was found in three <em>C. thous</em>. Phylogenetic analyses based on the near-complete 18S rRNA, <em>cox-1</em> and <em>hsp70</em> genes evidenced the occurrence of a novel of <em>Babesia</em> spp. (namely <em>Babesia pantanalensis</em> nov. sp.) closely related to <em>Rangelia vitalii</em> and <em>Babesia</em> sp. ‘Coco’. This finding was supported by the genetic divergence analysis which showed (i) high divergence, ranging from 4.17 to 5.62% for 18 S rRNA, 6.16% for <em>hps70</em> and 4.91–9.25% for <em>cox-1</em> and (ii) the genotype network (which displayed sequences separated from the previously described Piroplasmida species by median vectors and several mutational events). Also, phylogenetic analysis based on the 18S rRNA gene of <em>Hepatozoon</em> spp. positioned the sequences obtained herein in a clade phylogenetically related to <em>Hepatozoon</em> sp. ‘Curupira 2’, <em>Hepatozoon</em> sp. detected in domestic and wild canids from Uruguay and <em>Hepatozoon americanum</em>. The present study described <em>Babesia pantanalensis</em> nov sp. and <em>Hepatozoon</em> closely related to <em>H. americanum</em> in crab-eating foxes from Brazil. Moreover, the coinfection by piroplasmids and <em>Hepatozoon</em> sp. for the first time in crab-eating foxes strongly suggesting that this wild canid species potentially acts as a bio-accumulate of hemoprotozoan in wild environment.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"262 ","pages":"Article 108786"},"PeriodicalIF":2.1,"publicationDate":"2024-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140956802","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":"Genetic modification of the bee parasite Crithidia bombi for improved visualization and protein localization","authors":"Blyssalyn V. Bieber ,&nbsp;Sarah G. Lockett ,&nbsp;Sonja K. Glasser ,&nbsp;Faith A. St. Clair ,&nbsp;Neida O. Portillo ,&nbsp;Lynn S. Adler ,&nbsp;Megan L. Povelones","doi":"10.1016/j.exppara.2024.108789","DOIUrl":"10.1016/j.exppara.2024.108789","url":null,"abstract":"<div><p><em>Crithidia bombi</em> is a trypanosomatid parasite that infects several species of bumble bees (<em>Bombus</em> spp.), by adhering to their intestinal tract. <em>Crithidia bombi</em> infection impairs learning and reduces survival of workers and the fitness of overwintering queens. Although there is extensive research on the ecology of this host-pathogen system, we understand far less about the mechanisms that mediate internal infection dynamics. <em>Crithidia bombi</em> infects hosts by attaching to the hindgut via the flagellum, and one previous study found that a nectar secondary compound removed the flagellum, preventing attachment. However, approaches that allow more detailed observation of parasite attachment and growth would allow us to better understand factors mediating this host-pathogen relationship. We established techniques for genetic manipulation and visualization of cultured <em>C. bombi.</em> Using constructs established for <em>Crithidia fasciculata,</em> we successfully generated <em>C. bombi</em> cells expressing ectopic fluorescent transgenes using two different selectable markers. To our knowledge, this is the first genetic modification of this species. We also introduced constructs that label the mitochondrion and nucleus of the parasite, showing that subcellular targeting signals can function across parasite species to highlight specific organelles. Finally, we visualized fluorescently tagged parasites <em>in vitro</em> in both their swimming and attached forms, and <em>in vivo</em> in bumble bee (<em>Bombus impatiens</em>) hosts. Expanding our cell and molecular toolkit for <em>C. bombi</em> will help us better understand how factors such as host diet, immune system, and physiology mediate outcomes of infection by these common parasites.</p></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"262 ","pages":"Article 108789"},"PeriodicalIF":2.1,"publicationDate":"2024-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140956621","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}