Experimental parasitology最新文献

{"title":"Cystic echinococcosis susceptibility is increased by polymorphisms in the transporter associated with antigen processing 1 and 2 genes","authors":"Susan Jabbaripour ,&nbsp;Siamak Sandoghchian Shotorbani ,&nbsp;Adel Spotin ,&nbsp;Ehsan Ahmadpour ,&nbsp;Navid Shomali ,&nbsp;Mahmoud Mahami-Oskouei","doi":"10.1016/j.exppara.2025.108943","DOIUrl":"10.1016/j.exppara.2025.108943","url":null,"abstract":"<div><div>Susceptibility or resistance to various diseases, especially parasitic infections, largely depends on immune system function and genetics. Single nucleotide polymorphisms on the TAP1-637 and TAP2-379 genes may play a role in hydatidosis by affecting the immune system.</div><div>This study was conducted as a case-control study. After confirmation by serological ELISA method, blood samples were collected from 76 people with hydatid cyst and also from 76 healthy people. DNA was extracted from the samples and the ARMS-PCR technique was used to identify mutations. To evaluate the accuracy and ensure the PCR results, some samples were also sequenced in two groups with different genotypes.</div><div>Compared to the control group, the case group (infected group) had a higher frequency of the heterozygous Asp/Gly codon 637 of the TAP1 gene. In addition, the frequency of the Gly phenotype and allele was higher in the infected group (<em>P</em> &lt; 0.05). Furthermore, the infected group had a higher frequency of the heterozygous Val/Ile codon 379 of the TAP2 gene. The frequency of the Ile phenotype and allele was also higher in the infected group than in the control group (<em>P</em> &lt; 0. 05).</div><div>The results of this study showed that the Gly allele and Asp/Gly genotype in codon 637 of the TAP1 gene, as well as the Ile allele and Val/Ile genotype in codon 379 of the TAP2 gene are considered genetic risk factors for cystic echinococcosis.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"272 ","pages":"Article 108943"},"PeriodicalIF":1.4,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143917321","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":"Identification of therapeutics against PfPK6 protein of Plasmodium falciparum: Structure and Deep Learning approach","authors":"Sibasish Sarangi,&nbsp;Rajani Kanta Mahapatra","doi":"10.1016/j.exppara.2025.108947","DOIUrl":"10.1016/j.exppara.2025.108947","url":null,"abstract":"<div><div>The <em>Plasmodium falciparum</em> Protein Kinase 6 (PfPK6) is a serine/threonine protein kinase categorized under the CMGC group, displaying both cyclin-dependent kinases (CDKs) and mitogen-activated protein kinases (MAPKs) activity. Previous research has indicated that PfPK6 is expressed during the trophozoite and schizont stages of the <em>Plasmodium falciparum</em> asexual blood stage. Unlike typical cyclin-dependent kinases, PfPK6 demonstrates kinase activity independent of cyclin, making it a promising target for drug identification. In this study, we utilized a computational approach to identify a novel PfPK6 inhibitor through virtual screening of small inhibitor compounds from diverse datasets, employing a structure-based approach and a Deep Learning (DL) model. The most promising inhibitor molecule, TCMDC-132409 from the Tres Cantos Antimalarial Set, exhibited a binding affinity of −13.553 kcal/mol against PfPK6. Additionally, a 200ns molecular dynamics simulation study confirmed the stability of the binding mode, indicating the potential of TCMDC-132409 as an antiplasmodial inhibitor for further investigation.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"273 ","pages":"Article 108947"},"PeriodicalIF":1.4,"publicationDate":"2025-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143894559","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":"Molecular docking, toxicity study and in vitro antimalarial evaluation of pyrazole substituted 1,3,5-triazine derivatives","authors":"Bonita Chetia ,&nbsp;Anshul Shakya ,&nbsp;Surajit Kumar Ghosh ,&nbsp;Sathishkumar Vinayagam ,&nbsp;Saurav Jyoti Patgiri ,&nbsp;Ipsita Pal Bhowmick ,&nbsp;Udaya Pratap Singh ,&nbsp;Hans Raj Bhat","doi":"10.1016/j.exppara.2025.108945","DOIUrl":"10.1016/j.exppara.2025.108945","url":null,"abstract":"<div><div>The development of resistance to antimalarial drugs such as chloroquine, amodiaquine, artemisinin, and antifolates is a major health concern, prompting more research into new antimalarial therapies. In the present study, we intend to develop pyrazole substituted 1,3,5-triazine derivatives <strong>7</strong>(<strong>a</strong>-<strong>j</strong>) as antimalarial agents. These compounds were synthesized using conventional methods and analyzed using various spectroscopic techniques. The docking results showed that compounds <strong>7j</strong> and <strong>7i</strong> exhibited an excellent binding interactions with PRO A:113, ILE A:164, SER A:111, PHE A:58, LEU A:46, LEU A:119, VAL A:45, ILE A:112 (−204.97to −118.41 kcal/mol) and TYR A:170, CYS A:15, PRO A:113, ILE A:112, ALA A:16, PHE A:58, MET A:55, SER A:111 (−216.24to −152.06 kcal/mol) against wild (1J3I) and quadruple mutant (1J3K) type of <em>Pf</em>-DHFR-TS. Compounds <strong>7j</strong> and <strong>7i</strong> exhibited considerable antimalarial efficacy against chloroquine-sensitive (3D7) and chloroquine-resistant (Dd2) strains of <em>P. falciparum</em>, with IC₅₀ values ranging from 23.78 to 83.36 μM and 30.89–64.24 μM, respectively. These pyrazole-substituted 1,3,5-triazine derivatives could be utilized to find a novel class of <em>Pf</em>-DHFR-TS inhibitors.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"273 ","pages":"Article 108945"},"PeriodicalIF":1.4,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143881432","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":"Toll-like receptors and inflammatory cytokines in the skin of Acanthamoeba spp. infected immunocompetent and immunosuppressed mice","authors":"A. Wojtkowiak-Giera ,&nbsp;D. Kosik-Bogacka ,&nbsp;N. Łanocha-Arendarczyk ,&nbsp;A. Kolasa ,&nbsp;K. Kot ,&nbsp;P. Solarczyk ,&nbsp;M. Derda","doi":"10.1016/j.exppara.2025.108944","DOIUrl":"10.1016/j.exppara.2025.108944","url":null,"abstract":"<div><div><em>Acanthamoeba</em> spp. can cause opportunistic infections, such as cutaneous acanthamoebiasis (CA). Little is known about the role of TLRs and cytokines in the host skin during <em>Acanthamoeba</em> spp. infections. The study aimed to examine the gene and protein expression of TLR3, TLR7, IFN-γ, and IL-23 in the skin of mice experimentally infected with a clinical strain of <em>Acanthamoeba</em> spp. male BALB/c mice were assigned to four groups: group I (control group I) - with normal immunity (C, n = 5); group II (control group II) - with reduced immunity induced by methylprednisolone sodium succinate (MPS; CS, n = 5); group III - amoeba-infected hosts with normal immunity (A, n = 12); and group IV- amoeba-infected hosts with reduced immunity induced by MPS (AS, n = 12). The skin sections (2 cm × 2 cm) were collected from the animals at 8, 16, and 24 days post-infection (dpi). TLR3, TLR7, IFN-γ, and IL-23 gene and protein expressions were analyzed by quantitative real-time PCR and immunohistochemical staining.</div><div>In the immunocompetent hosts, we noted higher expressions of TLR3 and IL-23 at all-time points, except 8th dpi when IL-23 gene expression was downregulated compared to the control group. The mRNA expressions of TLR7 and IFN-γ were higher at 16 and 24 dpi in the skin of immunocompetent <em>Acanthamoeba</em> spp.-infected hosts than in the uninfected mice. In the course of acanthamoebiasis in the mice with reduced immunity, we found significant upregulation of TLR3, IL-23, and TLR7 gene expressions only at the beginning of infection compared to the control group. A similar relationship was observed for IFN-γ at 8 and 16 dpi.</div><div>The pathophysiology of <em>Acanthamoeba</em> infection in the skin is complex. The data presented in this paper add new insight, but they are not sufficient to explain the role of the studied receptors and cytokines. The clinical picture and mechanisms of host response appear to be influenced by the route of infection, immunological status and microorganisms carried within the parasites. CA remains a multifactorial phenomenon.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"273 ","pages":"Article 108944"},"PeriodicalIF":1.4,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143876988","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":"Application potential of arecoline hydrobromide for anti-Toxoplasma gondii","authors":"Xu Cheng ,&nbsp;Weifeng Yan ,&nbsp;Jingjie Xu ,&nbsp;Sihong Wang ,&nbsp;Chunmei Jin","doi":"10.1016/j.exppara.2025.108946","DOIUrl":"10.1016/j.exppara.2025.108946","url":null,"abstract":"<div><div>Although long-term chewing areca nut may cause cancer, it is very feasible to treat human or livestock diseases in a short period of time. In this study, arecoline hydrobromide (AH), which is extracted from <em>Areca catechu</em>, significantly inhibited the proliferation of tachyzoites of<em>Toxoplasma gondii</em> (<em>T. gondii</em>) <em>in vitro</em> and <em>in vivo</em>. Specially,<em>in vivo</em> AH effectively resisted liver damage originate from<em>T</em>. <em>gondii</em>. Furthemore, the survival and safety evaluation outcome indicated that AH can prolong survival period of mice infected with <em>T. gondii</em> at non-toxic and side-effect dose. Based on these advantages, AH is a candidate compound with great potential for the treatment of <em>T. gondii</em> infection and it may be used clinically in the future.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"273 ","pages":"Article 108946"},"PeriodicalIF":1.4,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143859483","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":"In silico screening and molecular dynamic simulations of FDA-approved drugs as an inhibitor of trypanothione reductase of Leishmania donovani","authors":"Pooja Beniwal ,&nbsp;Chandra Kanta Bhusal ,&nbsp;Gajendra Choudhary ,&nbsp;Rakesh Sehgal ,&nbsp;Bikash Medhi ,&nbsp;Ajay Prakash ,&nbsp;Sukhbir Kaur","doi":"10.1016/j.exppara.2025.108942","DOIUrl":"10.1016/j.exppara.2025.108942","url":null,"abstract":"<div><div>Visceral leishmaniasis (VL) is mainly caused by <em>Leishmania donovani (Ld) and Leishmania (L.) infantum,</em> and it is prevalent in Brazil, India and East Africa. VL is a serious health issue, affecting millions of people worldwide and causing thousands of deaths annually. The current treatments for leishmaniasis are inadequate because of their low efficacy, toxicity and growing resistance, underscoring the pressing need to explore new drugs.</div><div>Among the various molecular targets explored, trypanothione reductase (TR) is of special relevance because of its crucial function in regulating the parasite's redox homeostasis. Inhibiting TR can disrupt the redox homeostasis of the parasites, offering a promising strategy for developing new drugs with improved efficacy and safety profiles. In this study, 3D structure model of TR was elucidated by homology modelling and potential novel inhibitors against <em>Leishmania donovani</em> TR (<em>Ld</em>TR) were identified by performing high-throughput virtual screening of 1615 FDA-approved drugs from the ZINC database <em>via</em> molecular docking, selecting top ligands on the basis of their high binding score and number of hydrogen bonds. These best hits are further subjected to Molecular Dynamics (MD) simulation and Molecular Mechanics Poisson- Boltzmann Surface Area (MM-PBSA) analysis. The results indicated that the binding scores of Dasatinib, Regorafenib, Bicalutamide, Raloxifene and Silodosin are −10.9 and −10.6, −10.1, −9.7 and −9.6 kcal/mol respectively. The lead compounds i.e. Dasatinib, Regorafenib, Bicalutamide, Raloxifene and Silodosin complexes with our target TR were found to be stable during MD simulation studies. Furthermore, MM-PBSA analysis demonstrated that these compounds had a high negative binding free energy. Thus, <em>in-silico</em> results showed that Dasatinib, Regorafenib, Bicalutamide and Raloxifene and Silodosin seem to have efficacy against TR for the treatment of VL. With further <em>in vitro</em> and <em>in vivo</em> investigations Dasatinib, Regorafenib, Bicalutamide, Raloxifene and Silodosin could be a good candidate of choice for combating leishmaniasis.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"272 ","pages":"Article 108942"},"PeriodicalIF":1.4,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742708","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":"Evaluation of the resistance of Anisakis spp. larvae to products regularly used in the industry and households","authors":"Abdelkader Biary ,&nbsp;Salma Berrouch ,&nbsp;Brahim Mimouni ,&nbsp;Jamal Eddine Hafid","doi":"10.1016/j.exppara.2025.108932","DOIUrl":"10.1016/j.exppara.2025.108932","url":null,"abstract":"<div><div>The rise in popularity of raw or undercooked fish dishes like sushi, sashimi, and ceviche has led to an increase in human cases of anisakiasis. This study aimed to evaluate the resistance of Anisakis larvae to substances like bleach, salt, brine, and marinades. Batches of 10 larvae were exposed to various concentrations of commercial bleach (pure, 50 %, 25 %, 12.5 %, 6.25 %) and salt (dry, 360 g/L, 90 g/L, 45 g/L, 20 g/L). Larvae were also tested with two vinaigrettes (one with mustard and one without) and four marinades made with vinegar and salt. Sodium hypochlorite was highly effective, killing larvae within minutes, with pure bleach eliminating them in 40 s. Dry salt killed them in under 70 min, while different concentrations of brine inactivated them over a period ranging from 3 to 17 days. Dressings with mustard worked faster than those without, and vinegar with higher acetic acid content (8 %) killed larvae in 6 h, compared to 29 h for 6 % acetic acid. Salt and vinegar marinades were more effective together: a combination of 8 % acetic acid and 6 % salt killed larvae in 2.5 h. Overall, sodium hypochlorite proved to be the most effective, while the combination of salt and vinegar also significantly reduced larval survival. These findings highlight the importance of using bleach for disinfecting surfaces and utensils after handling fish to prevent cross-contamination and ensure food safety.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"272 ","pages":"Article 108932"},"PeriodicalIF":1.4,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143669366","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":"Suppression of 8-oxoguanine DNA glycosylase (OGG1) activity produced positive impacts on disease severity, survival, and histopathological features of mice infected with Plasmodium berghei","authors":"Mukhtar Gambo Lawal ,&nbsp;Abdullahi Samaila ,&nbsp;Rusliza Basir ,&nbsp;Nur Aimi Liyana Abd Aziz ,&nbsp;Abdusalam Abdullah Alarabei ,&nbsp;Maizaton Atmadini Abdullah ,&nbsp;Roslaini Abd Majid ,&nbsp;Norshariza Nordin ,&nbsp;Mohd Khairi Hussain ,&nbsp;Elysha Nur Ismail","doi":"10.1016/j.exppara.2025.108930","DOIUrl":"10.1016/j.exppara.2025.108930","url":null,"abstract":"<div><div>Malaria is a life-threatening disease, leading to significant morbidity and mortality. Malaria treatment remains a challenge due to its intricate pathophysiology and high levels of parasite resistance to many currently available antimalarial agents. Thus, there is an urgent need for more therapeutic strategies to combat the disease. OGG1 activity has been implicated in many inflammatory disease conditions, making suppressing OGG1 activity a potential target for therapeutic purposes. The current study aimed to determine the effect of suppressing OGG1 activity on the severity, survival, and histopathological features of <em>P. berghei</em>-infected mice. In this study, the effects of modulating OGG1 activity on parasitaemia development, disease progression, survival rate, and histopathological outcomes in major organs of <em>Plasmodium berghei</em> (<em>P. berghei)</em> infected mice were evaluated. A significant difference in the mean parasitaemia was observed between the Vehicle, TH5487-treated, and O8-treated mice (p &lt; 0.001). Vehicle-treated mice exhibited markedly elevated mean percentage parasitaemia and succumbed to the infection earlier than TH5487 and O8-treated mice. The O8-treated mice showed the highest parasitaemia reduction of 39.60 ± 1.53 % compared to TH5487-treated mice. Histopathological examination revealed less severe pathological features associated with <em>P. berghei</em> infection in mice treated with OGG1 inhibitors than in vehicle-treated malaria mice. Significant differences were observed in the sequestration of PRBC, inflammation, hemozoin deposition, and architectural loss in mice treated with O8 and TH5487 compared to untreated malaria mice. The results of this study suggested that OGG1 suppression led to a decrease in parasitaemia and severity of the histopathological features in <em>P. berghei</em>-infected mice. The increased survival of treated malaria mice further supported this effect. These findings indicate that OGG1 suppression could be a potential therapeutic strategy during malaria.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"272 ","pages":"Article 108930"},"PeriodicalIF":1.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633987","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":"Involvement of Toxoplasma gondii natural antisense transcripts in cellular stress responses","authors":"Yue Gou,&nbsp;Laura Agudelo Vallejo,&nbsp;Ana Podadera,&nbsp;Kenneth Ng,&nbsp;Sirinart Ananvoranich","doi":"10.1016/j.exppara.2025.108931","DOIUrl":"10.1016/j.exppara.2025.108931","url":null,"abstract":"<div><div>Natural antisense transcripts (NATs), as a major subset of long non-coding RNAs (lncRNAs), are derived from every chromosome of <em>Toxoplasma gondii</em>, with the highest occurrence from ChrIa (18.4 NATs per Mbp) and the lowest from ChrIX (3.9 NATs per Mbp). GO analysis indicates that genes, which mRNA-NAT pairs are derived, are important for house-keeping and essential activities of <em>T. gondii</em>. Approximately half of protein encoding genes, whose loci also generate NATs, are involved in biological processes of metabolic processes and protein biochemistry and have canonical catalytic or binding activities. Using NAT of ubiquitin-like protease 1 (<em>TgUlp1</em>-NAT) as our study model, we showed that <em>TgUlp1</em>-NAT expression is part of cellular stress responses. Using a nanoluc reporter system, we confirmed that electroporation or membrane destabilization significantly induced <em>TgUlp1</em>-NAT expression. When the extracellular parasites were exposed to media containing high potassium, high sodium or altered osmotic pressure, <em>TgUlp1</em>-NAT expression was significantly down-regulated. In addition, two <em>TgUlp1</em>-NAT variants were detected in stressed <em>T. gondii</em>. One is an intron-retained variant, and the other is a spliced variant, referred to as <em>TgUlp1</em>-NATa and <em>TgUlp1</em>-NATb, respectively. The intronic sequence is 368 nts long, where regulatory small ncRNAs were derived. Taken together, we have confirmed that NAT expressions and functions are involved in cellular adaptation that allows <em>T. gondii</em> recover from stresses.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"271 ","pages":"Article 108931"},"PeriodicalIF":1.4,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143630281","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":"Development of a multiplex PCR assay targeting mitochondrial cytochrome oxidase subunit III (cox3) gene for simultaneous specific and sensitive detection of Babesia gibsoni and Babesia vogeli in dogs","authors":"Mitesh Mittal ,&nbsp;Soumendu Chakravarti ,&nbsp;Krishnendu Kundu ,&nbsp;Prashant Tripathi ,&nbsp;Pramod Batra","doi":"10.1016/j.exppara.2025.108922","DOIUrl":"10.1016/j.exppara.2025.108922","url":null,"abstract":"<div><div>Canine babesiosis is a potential threat for the dog population worldwide. Rapid, sensitive, and specific identification of the etiological agent to the species is pivotal for initiating effective therapeutic and control measures. Co-infection with multiple species pathogens due to multiple vectors infesting dogs is not uncommon. A multiplex PCR (Bg-Bv mPCR) for simultaneous detection and differentiation of the two common <em>Babesia</em> species, <em>B. gibsoni</em> and <em>B</em>. <em>vogeli</em> has been developed targeting the mitochondrial cytochrome oxidase subunit III (cox3) gene. These two species are the species responsible for causing canine babesiosis in Indian subcontinent and Southern Asia. This cox3 gene is present in high copy number and the sequences are species specific and hence targeted to develop the diagnostic multiplex PCR. The multiplex PCR was able to detect up to 5 pg DNA of the <em>Babesia</em> species. No cross-amplifications were observed between the primers specific for either <em>B. vogeli</em> or <em>B. gibsoni</em>. The Bg-Bv mPCR resulted in significantly higher <em>B. gibsoni</em> positives (30/250) than existing 18S ribosomal RNA PCR (22/250). Similarly, the mPCR detected more <em>B. vogeli</em> (26/250) than the 18S rRNA PCR (18/250). The kappa statistics when applied to the results generated by each of the PCR tests also revealed a substantial to perfect agreement between the data. The multiplex PCR targeting <em>cox3</em> gene is thus a rapid, sensitive, and specific method for simultaneous detection and differentiation of the <em>B</em>. <em>gibsoni</em> and <em>B</em>. <em>vogeli</em>.</div></div>","PeriodicalId":12117,"journal":{"name":"Experimental parasitology","volume":"271 ","pages":"Article 108922"},"PeriodicalIF":1.4,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143585295","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}