Experimental parasitology最新文献_第2页

IgG avidity and placenta real-time PCR in detection of active maternal toxoplasmosis: relation to pregnancy outcomes IgG亲和度及胎盘实时PCR检测活动性母体弓形虫病：与妊娠结局的关系。

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-12-01 DOI: 10.1016/j.exppara.2025.109065

Amal Farahat Allam , Amel Youssef Shehab , Nashwa Abdelaleem Ahmed , Amel Gaber Elshereedy , Hoda Fahmy Farag , Sarah Mohamed Abdo , Heba Said Ibrahim

Toxoplasmosis diagnosis during pregnancy is important for the management of pregnant women suspected to have early T. gondii infection. This study aimed to detect T. gondii infection using ELISA and placenta real-time PCR among pregnant women. The study involved 149 women from El Shatby Hospital, Alexandria University; 50 experienced spontaneous abortion and 99 delivered normally, among whom four cases were spiramycin treated. Only 83 women agreed to submit blood samples that were ELISA tested for Toxoplasma IgG and IgM antibodies (ELISA Biokit, Barcelona, Spain). ELISA IgG positives were re-examined for IgG avidity. Placental samples were collected from all participants (about 20 g from each), and DNA was extracted using the Qiagen DNA kit, Hilden, Germany. The samples were examined by real-time PCR targeting the REP-529 gene. Out of the 83 women, 57 (68.7 %) were IgG positive and only one case was IgM positive. Fifty IgG cases had low IgG avidity and seven had high IgG avidity. Approximately one-third of the 149 placenta samples, 83 serologically examined cases, 57 IgG positives; 50 with low avidity, and 7 with high avidity, tested positive by real-time PCR, with detection rates of 29.5 %, 28.9 %, 31.5 %, 32 %, and 28.6 %, respectively. Moreover, it detected six positives among the IgG negatives, and two of the four spiramycin-treated cases tested positive. No significant difference between abortion and normal delivery rates was observed among T. gondii positive and negative women, either by ELISA IgG and/or real-time PCR. In conclusion, combining IgG avidity and placenta real-time PCR is promising for detecting T. gondii active infection and the probability of fetal infection.

妊娠期弓形虫病诊断对于怀疑早期弓形虫感染的孕妇的管理很重要。采用ELISA和胎盘实时荧光定量PCR检测孕妇弓形虫感染情况。这项研究涉及来自亚历山大大学El Shatby医院的149名妇女；自然流产50例，正常分娩99例，其中螺旋霉素治疗4例。只有83名妇女同意提交经ELISA检测弓形虫IgG和IgM抗体的血液样本（ELISA Biokit，巴塞罗那，西班牙）。再次检测ELISA IgG阳性。收集所有参与者的胎盘样本（每人约20 g），并使用Qiagen DNA试剂盒提取DNA， Hilden，德国。采用real-time PCR检测REP-529基因。83例女性中IgG阳性57例（68.7%），IgM阳性1例。IgG低贪婪50例，高贪婪7例。149份胎盘样本中约有三分之一，血清学检查病例83例，IgG阳性57例；实时荧光定量PCR检测阳性的低亲和度为50份，高亲和度为7份，检出率分别为29.5%、28.9%、31.5%、32%和28.6%。此外，它在IgG阴性中检测到6例阳性，并且在4例使用螺旋霉素治疗的病例中检测出2例阳性。弓形虫阳性和阴性妇女的流产率和正常分娩率在ELISA IgG和/或实时PCR检测中均无显著差异。综上所提，结合IgG亲和度和胎盘实时荧光定量PCR检测弓形虫活动性感染和胎儿感染的可能性是有希望的。

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引用次数: 0

Comparison of IgM and IgG ELISAs using recombinant thrombospondin-related adhesive protein (BgTRAP) for the differentiation of early and late Babesia gibsoni infections in canines 重组血小板反应相关黏附蛋白（BgTRAP） IgM和IgG elisa鉴别犬早期和晚期巴贝斯虫感染的比较

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-12-01 DOI: 10.1016/j.exppara.2025.109066

Nellikka Chalapron Sreenidhi , Chundayil Kalarickal Deepa , Anju Varghese , Karapparambu Gopalan Ajith Kumar , Aravindakshan Anaswara , Reghu Geethu , Ravindran Rajasekhar , Pulikottil Vinu David , Reghu Ravindran

Vector-borne illnesses pose a rising global threat to pet health, with Babesia gibsoni being the most common haemoprotozoan infection in canines of South India. Low parasitaemia either in early or late infection is a big hurdle in the diagnosis of canine babesiosis. The present study aimed to differentiate the early and late B. gibsoni infections in dogs using IgM/IgG ELISA based on recombinant thrombospondin-related adhesive protein (BgTRAP). The N-terminal BgTRAP gene was cloned into pET32a, expressed in BL21 Escherichia coli cells, and purified to get the recombinant protein. Using the recombinant antigen, IgM ELISA detected anti-B. gibsoni IgM antibodies in 73 out of 130 samples (56.15 per cent) while the IgG ELISA detected IgG antibodies in 77 out of 130 samples (59.23 per cent). Polymerase chain reaction of the 130 samples targeting the BgTRAP gene revealed 73.07 % positivity. When compared with the PCR, the sensitivity and a specificity of newly standardized indirect IgM ELISA were 36.54 % and 75.00 % while that of IgG ELISA were 42.31 % and 71.88 %. The accuracy, positive predictive value and negative predictive value of the IgM ELISA in comparison to PCR were 51.19 %, 70.4 %, 42.1 % while that of IgG ELISA were 53.57 %, 71.0 %, 43.4 % respectively. The area under the curve (AUC) and Youden Index in ROC curve for IgM and IgG ELISAs revealed a moderate diagnostic accuracy. The rBgTRAP antigen showed no cross-reactivity with common helminth parasites viz, Ancylostoma caninum, Dirofilaria immitis, D. repens, Spirometra spp., Toxocara canis and haemoparasites like Trypanosoma evansi, B. vogeli, Hepatozoon canis and Ehrlichia canis. The assay could clearly differentiate early (IgM) and late (IgG) infections, making it a valuable diagnostic tool for differentiating early and late infections with B. gibsoni.

媒介传播的疾病对宠物健康构成了日益严重的全球性威胁，而巴贝斯虫是南印度犬类中最常见的原虫感染。早期或晚期感染的低寄生血症是犬巴贝斯虫病诊断的一大障碍。本研究旨在利用基于重组血栓反应蛋白相关黏附蛋白（BgTRAP）的IgM/IgG酶联免疫吸附试验（ELISA）鉴别犬早期和晚期gibsoni感染。将n端BgTRAP基因克隆到pET32a中，在BL21大肠杆菌细胞中表达，纯化得到重组蛋白。采用重组抗原，IgM ELISA检测抗- b。130份样本中有73份（56.15%）检测到gibsoni IgM抗体，IgG ELISA检测到77份（59.23%）IgG抗体。130份BgTRAP基因的聚合酶链反应阳性率为73.07%。与PCR比较，新标准化IgM间接ELISA检测的灵敏度和特异性分别为36.54%和75.00%，IgG ELISA检测的灵敏度和特异性分别为42.31%和71.88%。IgM ELISA与PCR的准确率、阳性预测值、阴性预测值分别为51.19%、70.4%、42.1%，IgG ELISA的准确率分别为53.57%、71.0%、43.4%。IgM和IgG elisa的曲线下面积（AUC）和ROC曲线上的约登指数显示出中等的诊断准确性。rBgTRAP抗原与常见寄生虫如犬钩虫、免疫dirofilia、repens、螺虫、犬弓形虫和伊文氏锥虫、沃氏巴贝虫、犬肝虫、犬埃利希体等无交叉反应性。该方法可明确区分早期（IgM）和晚期（IgG）感染，是区分早期和晚期感染的有价值的诊断工具。

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引用次数: 0

Phytochemical-based discovery of a potent antimalarial candidate targeting PfPI4K: A hybrid structure-based and deep learning approach 基于植物化学的有效抗疟疾候选药物PfPI4K靶向的发现：基于结构和深度学习的混合方法。

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-12-01 DOI: 10.1016/j.exppara.2025.109064

Sibasish Sarangi, Rajani Kanta Mahapatra

Malaria is a parasitic infection that poses a threat to life and continues to be a serious challenge to global health. The recent COVID-19 pandemic has further escalated the situation. The emergence of artemisinin partial resistance and insecticide resistance emphasizes the critical need for novel antimalarial drug targets and agents with alternative mechanisms of action. This study focuses on Plasmodium falciparum phosphatidylinositol 4-kinase (PfPI4K), a phosphoinositide lipid kinase essential for membrane trafficking and biogenesis across multiple stages of the Plasmodium life cycle. We investigated a dataset of 58 natural anthraquinones with reported antimalarial activity as potential PfPI4K inhibitors. Employing Modeller 10.5 for homology modeling, we constructed the PfPI4K structure, validated by quality testing parameters. Subsequent in silico screening identified potential drug candidates. The top-scoring inhibitors were investigated by ADMET analysis. The compound AD37 (6′-O-methyl-knipholone) was identified as a prominent candidate. It complied with Lipinski's rule of five, displayed favorable ADMET parameters, and reported the highest binding affinity of −5.983 kcal/mol to PfPI4K as determined by GLIDE analysis. The stability and molecular interactions of the PfPI4K-AD37 complex were further confirmed by a 100 ns molecular dynamics simulation employing GROMACS. This investigation identifies AD37 as a promising drug candidate for treating malaria and provides valuable information regarding the molecular interactions essential for the future design and development of antimalarial drugs.

疟疾是一种寄生虫感染，对生命构成威胁，并继续对全球健康构成严重挑战。最近的COVID-19大流行使局势进一步升级。青蒿素部分耐药和杀虫剂耐药的出现强调了迫切需要具有替代作用机制的新型抗疟药物靶点和制剂。本研究的重点是恶性疟原虫磷脂酰肌醇4-激酶（PfPI4K），这是一种磷脂酰肌醇脂激酶，对疟原虫生命周期多个阶段的膜运输和生物发生至关重要。我们调查了58种天然蒽醌类药物的数据集，这些药物被报道为潜在的PfPI4K抑制剂，具有抗疟疾活性。采用modelmodel10.5进行同源性建模，构建了PfPI4K结构，并通过质量测试参数进行了验证。随后的计算机筛选确定了潜在的候选药物。通过ADMET分析对得分最高的抑制剂进行了研究。化合物AD37（6′-O-methyl-knipholone）被认为是一个突出的候选化合物。它符合Lipinski的五法则，具有良好的ADMET参数，经GLIDE分析，对PfPI4K的结合亲和力最高，为-5.983 kcal/mol。采用GROMACS进行100 ns分子动力学模拟，进一步证实了PfPI4K-AD37复合物的稳定性和分子相互作用。这项研究确定了AD37作为治疗疟疾的有前途的候选药物，并为未来抗疟疾药物的设计和开发提供了重要的分子相互作用信息。

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引用次数: 0

Therapeutic potential of spiramycin-nanoparticles and Aluvia in experimental congenital toxoplasmosis 螺旋霉素纳米颗粒和Aluvia治疗实验性先天性弓形虫病的潜力。

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-12-01 DOI: 10.1016/j.exppara.2025.109069

Yasmen Elsayed , Amal Farahat Allam , Amel Youssef Shehab , Naglaa Fathi Abd El-Latif , Shaimaa Makled , Nermine Mogahed Fawzy Hussein Mogahed

This study aimed to evaluate the efficacy of spiramycin, spiramycin-loaded chitosan nanoparticles (CSNPs) and Aluvia for the treatment of congenital toxoplasmosis in a murine model. The study was conducted on 60 pregnant Swiss albino mice, 30 controls and 30 experimental. After performing a pilot study, mice were injected subcutaneously with the virulent Toxoplasma gondii RH strain (30 tachyzoites/mouse) on day 15 of pregnancy (3rd gestation period). The drugs were evaluated based on pregnancy outcomes (number of mice with live birth, stillbirth and abortion), number of live offspring, live pups' weight and congenital anomalies. Histopathological changes in the offspring's brain were studied. Regarding pregnancy outcomes, all mice in the non-infected control group delivered live offspring. T. gondii infection significantly decreased the live birth rate, while treatment with spiramycin, spiramycin-loaded CSNPs, and Aluvia improved pregnancy outcomes without statistical significance compared to the non-infected control. The highest number of offspring was observed in the normal non-infected control subgroup (94 pups/10 mice). Among the treated groups, spiramycin-loaded CSNPs resulted in the highest offspring count (46 pups/10 mice) and ranked first in mean pups weight, followed by Aluvia. Congenital anomalies were observed among the offspring of both infected untreated and infected treated mothers. Offspring brain tissues revealed substantial histopathological improvement in the spiramycin-loaded CSNPs and Aluvia-treated groups. In conclusion, spiramycin-loaded CSNPs and Aluvia demonstrated low parasite burden and successfully restored normal brain architecture in the offspring. However, congenital anomalies persisted and remained a significant concern.

本研究旨在评价螺旋霉素、螺旋霉素负载壳聚糖纳米颗粒（csnp）和Aluvia治疗小鼠先天性弓形虫病的疗效。研究对象为60只怀孕的瑞士白化病小鼠、30只对照小鼠和30只实验小鼠。在进行初步研究后，小鼠在妊娠第15天（第三妊娠期）皮下注射毒性强的弓形虫RH株（30个速殖子/只）。根据妊娠结局（活产鼠数、死产鼠数、流产鼠数）、活仔鼠数、活仔鼠体重和先天性异常情况对药物进行评估。研究了后代大脑的组织病理学变化。关于妊娠结局，未感染对照组的所有小鼠都产下了活的后代。弓形虫感染显著降低了活产率，而螺旋霉素、螺旋霉素负载csnp和Aluvia治疗改善了妊娠结局，但与未感染的对照组相比，无统计学意义。正常未感染对照组的子代数量最多（94只/10只）。在处理组中，携带螺旋霉素的csnp导致最高的后代数量（46只/10只小鼠），平均幼崽体重排名第一，其次是Aluvia。在感染未治疗和感染治疗的母亲的后代中都观察到先天性异常。在螺旋霉素加载csnp和aluvia处理组中，后代脑组织显示出实质性的组织病理学改善。综上所述，携带螺旋霉素的csnp和Aluvia表现出较低的寄生虫负担，并成功地恢复了后代正常的大脑结构。然而，先天性异常持续存在，仍然值得关注。

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引用次数: 0

Axenic culture of Entamoeba histolytica alters its ability to interact with pathogenic bacteria and leads to loss of virulence and pathogenicity 溶组织内阿米巴的无菌培养改变了其与致病菌相互作用的能力，并导致毒性和致病性的丧失

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-12-01 DOI: 10.1016/j.exppara.2025.109072

Fabrício Marcus Silva Oliveira , Sávio Henrique de Cicco Sandes , Mario Abatemarco Junior , Elisabeth Neumann , Álvaro Cantini Nunes , Maria Aparecida Gomes , Marcelo Vidigal Caliari

Entamoeba histolytica is an anaerobic eukaryotic protozoan capable of infecting humans and causing amoebiasis, a disease responsible for approximately 50 million cases and an estimated 100,000 deaths annually worldwide. The virulence of Entamoeba histolytica trophozoites and their ability to cause amoebic colitis and hepatic abscesses in the host involve various molecules, such as a specific lectin that recognizes galactose and N-acetylgalactosamine residues, cysteine proteinases, and amoebapores. To determine whether axenization of the Entamoeba histolytica strain affects its ability to interact with and respond to the presence of other microorganisms, we cultured trophozoites of an Entamoeba histolytica EGG strain in both xenic and axenic media. We then associated them with Salmonella typhimurium and evaluated the expression of virulence factors by real-time PCR. Subsequently, we infected rats to assess whether the Enatmoeba histolytica EGG strain cultured under xenic and axenic conditions exhibited differences in pathogenicity. This study showed that the absence of bacterial microbiota in axenic culture led to a reduced capacity to produce virulence factors in vitro, even when stimulated with Salmonella typhimurium. Furthermore, in vivo analysis revealed that axenic culture altered the parasite's behavior by reducing its ability to produce intestinal lesions, even in a co-infection setting. We suggest that the expression profiles of virulence genes are influenced by continuous environmental stress and that the presence of microbiota in xenic cultures of Enatmoeba histolytica contributes to such stress and supports the maintenance of virulence gene expression.

溶组织内阿米巴原虫是一种厌氧真核原生动物，能够感染人类并引起阿米巴病，阿米巴病每年在全世界造成约5000万例病例和估计10万人死亡。溶组织内阿米巴滋养体的毒力及其在宿主中引起阿米巴结肠炎和肝脓肿的能力涉及各种分子，如识别半乳糖和n -乙酰半乳糖胺残基的特定凝集素、半胱氨酸蛋白酶和阿米巴孢子。为了确定溶组织内阿米巴菌株的无菌化是否会影响其与其他微生物相互作用和反应的能力，我们在xenic和xenic培养基中培养了溶组织内阿米巴卵细胞菌株的滋养体。然后，我们将它们与鼠伤寒沙门菌联系起来，并通过实时PCR评估毒力因子的表达。随后，我们感染了大鼠，以评估在无菌和无菌条件下培养的溶组织肠内巴卵细胞菌株是否具有不同的致病性。这项研究表明，无菌培养中细菌微生物群的缺乏导致体外产生毒力因子的能力降低，即使用鼠伤寒沙门菌刺激也是如此。此外，体内分析显示，即使在共同感染环境中，无菌培养也会通过降低其产生肠道病变的能力来改变寄生虫的行为。我们认为，毒力基因的表达谱受到持续环境胁迫的影响，而溶组织肠杆菌异种培养中微生物群的存在有助于这种胁迫，并支持毒力基因表达的维持。

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引用次数: 0

A predictive model for predicting the occurrence of liver cancer in patients with schistosomiasis cirrhosis 血吸虫病肝硬化患者肝癌发生的预测模型

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-12-01 DOI: 10.1016/j.exppara.2025.109071

Yiming Wu , Jing Huang , Chenzhou Xu

Objective

Early diagnosis of liver cancer following schistosomiasis cirrhosis remains challenging. This study aims to develop a predictive model for identifying precancerous liver lesions and early-stage liver cancer in patients with schistosomiasis cirrhosis.

Methods

Patients diagnosed with schistosomiasis cirrhosis over the past 14 years and managed by the Jiaxing Schistosomiasis Management Office were included in the study. Univariate regression analysis was conducted to assess the correlation between various indices and liver cancer. The predictive diagnostic efficiency was evaluated using the AUROC's cross-sectional area, with the optimal cutoff point identified through the Youden index.

Results

Univariate regression revealed significant associations with RBC count, ALT level, and other factors (P<0.1). Multivariate regression analysis identified AST, GGT, and additional factors as significant predictors (P<0.05). From these findings, two predictive models were developed: one for early-stage liver cancer (ESLC model) and another for precancerous lesions (PL model). The AUROC confirmed the superior diagnostic performance of both models compared to AFP, particularly in predicting precancerous lesions, thus addressing some of AFP's limitations.

Conclusion

The proposed predictive model serves as a valuable tool for early detection of precancerous liver lesions. It outperforms AFP in this regard, with AST, GGT, TB, BA, PT, PCIII, and AFP identified as independent predictors of liver cancer.

目的血吸虫病肝硬化后肝癌的早期诊断仍然具有挑战性。本研究旨在建立血吸虫病肝硬化患者癌前病变和早期肝癌的预测模型。方法选取嘉兴市血吸虫病管理办公室管理的14年内确诊为肝硬化的患者为研究对象。采用单因素回归分析评价各指标与肝癌的相关性。使用AUROC的横截面积评估预测诊断效率，并通过约登指数确定最佳截止点。结果单因素回归显示与RBC计数、ALT水平及其他因素有显著相关性（P<0.1）。多因素回归分析发现AST、GGT和其他因素是显著的预测因子（P<0.05）。根据这些发现，开发了两种预测模型：一种用于早期肝癌（ESLC模型），另一种用于癌前病变（PL模型）。AUROC证实，与AFP相比，这两种模型的诊断性能更好，特别是在预测癌前病变方面，从而解决了AFP的一些局限性。结论该预测模型可作为早期发现肝癌前病变的有效工具。在这方面，它优于AFP， AST、GGT、TB、BA、PT、PCIII和AFP被确定为肝癌的独立预测因子。

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引用次数: 0

Infection dynamics of Metarhizium anisopliae in Rhipicephalus microplus and the role of tick traits in susceptibility 金龟子绿僵菌在微小鼻头虫中的感染动态及蜱虫性状对其易感性的影响。

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-11-21 DOI: 10.1016/j.exppara.2025.109070

Nurul Fatin Amirah Mohd Azmi , Samsuddin Ahmad Syazwan , Ruhil Hayati Hamdan , Tan Li Peng

Tick-borne diseases (TBDs) pose a major threat to global livestock production, particularly in tropical and subtropical regions, and their burden is projected to increase worldwide in the coming decades as a result of climate change. In Brazil, tick infestations are responsible for economic losses estimated at about USD 3.24 billion annually. Growing resistance to chemical acaricides necessitates alternative control strategies such as biological agents. This study assessed the pathogenicity of a local strain of Metarhizium anisopliae (PR1) against adult Rhipicephalus microplus ticks using the Adult Immersion Test at conidial concentrations of 10⁷, 10⁸, and 10⁹ conidia/mL, each tested in triplicate alongside positive and negative controls. All concentrations caused complete tick mortality within 14 days, with higher concentrations accelerating tick death and completely inhibiting oviposition and larval hatchability. The estimated LT₅₀ and LT₉₀ values for the 10⁹ conidia/mL treatment were 2.46 [2.09–2.75] days and 3.63 [3.25–4.29] days, respectively. In contrast, untreated ticks exhibited only 6.7 % mortality and laid an average of 1567 ± 314 eggs, of which 84 % hatched. A significant positive correlation was observed between tick weight and the time to death at the highest concentration, suggesting heavier ticks resisted infection longer, potentially due to higher energy reserves or thicker cuticular. These results underline the potent in vitro pathogenic and reproductive-suppressing effects of M. anisopliae strain PR1 under laboratory conditions, while emphasizing the need for further field validation before considering its application in broader vector management programs.

蜱传疾病（tbd）对全球畜牧业生产构成重大威胁，特别是在热带和亚热带地区，预计未来几十年，由于气候变化，其负担将在全球范围内增加。在巴西，蜱虫侵扰每年造成的经济损失估计约为32.4亿美元。对化学杀螨剂的抗药性日益增强，需要采用生物制剂等替代控制策略。本研究采用成虫浸没试验，在分生孢子浓度分别为107、108和109分生孢子/mL的情况下，对当地一株金银绿僵菌（PR1）对微头蜱成虫的致病性进行了评估。各浓度均能使蜱在14 d内完全死亡，浓度越高，则加速了蜱的死亡，并完全抑制了其产卵和幼虫的孵化能力。109个分生孢子/mL处理的LT50和LT90分别为2.46[2.09-2.75]天和3.63[3.25-4.29]天。相比之下，未经处理的蜱死亡率仅为6.7%，平均产卵1567±314枚，其中84%孵化。在最高浓度下，蜱的体重与死亡时间呈显著正相关，表明较重的蜱对感染的抵抗力较长，可能是由于能量储备较高或表皮较厚。这些结果强调了绿僵菌PR1在实验室条件下具有强大的体外致病和生殖抑制作用，同时强调了在考虑将其应用于更广泛的媒介管理计划之前需要进一步的现场验证。

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引用次数: 0

Evaluation of the efficacy of four fasciolicides in naturally infected cattle in Beni-Suef governorate, Egypt: First report of therapeutic failure of triclabendazole 埃及贝尼-苏韦夫省四种杀片虫剂对自然感染牛的疗效评价：首次报道三氯咪唑治疗失败。

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-11-21 DOI: 10.1016/j.exppara.2025.109067

Mohamed G. Abdelrahman , Shawky M. Aboelhadid , Sherin R. Rouby , Ahmed Zaitoun , Ahmed M. El-Sherif

The emergence of anthelmintic resistance presents a major threat to the health and productivity of grazing livestock. This study aimed to evaluate the efficacy of commonly used fasciolicides against Fasciola spp. in naturally infected cattle from Beni-Suef, Egypt—an area where fasciolosis is endemic. Twenty adult female cattle, confirmed by microscopic examination to be infected with Fasciola spp., were selected and divided into three experimental trials to evaluate the efficacy of four commercial fasciolicides: triclabendazole (TCBZ), rafoxanide (RAF), nitroxynil (NIT), and oxyclozanide (OCZ). Experiment I evaluated possible resistance to TCBZ, RAF, and NIT by administering two consecutive doses of each drug. Experiment II investigated potential multidrug resistance by treating animals resistant to RAF and NIT with a single dose of TCBZ. Experiment III examined the efficacy of OCZ as an alternative treatment. The fecal egg count reduction test (FECRT) was used as the primary assessment method in all trials. In Experiment I, cattle treated with RAF or NIT showed no reduction in egg counts, even after the second dose. TCBZ treatment resulted in a marked decrease in egg counts at day 7 post-treatment, but counts gradually increased from day 14 onwards, returning to pre-treatment levels by day 56. In Experiment II, animals treated with TCBZ again showed a significant drop in egg counts at day 7, followed by a steady rise to baseline values by day 28. In Experiment III, OCZ achieved complete elimination of eggs by day 7, with 100 % efficacy. This study provides the first documented evidence of triclabendazole (TCBZ) treatment failure in animals in Egypt, highlighting the urgent need for large-scale investigations across the country. In contrast, oxyclozanide (OCZ) showed high efficacy in eliminating Fasciola infections in cattle, emphasizing its potential as a valuable alternative for improving fasciolosis control and management strategies in Egypt.

抗虫性的出现对放牧牲畜的健康和生产力构成了重大威胁。本研究旨在评估常用的杀片形虫剂对埃及贝尼-苏韦夫自然感染的牛片形虫的疗效。贝尼-苏韦夫是片形虫病流行的地区。选取镜检证实感染片吸虫病的成年母牛20头，分为3个试验，分别评价三氯苄达唑（TCBZ）、拉福昔尼（RAF）、硝基尼（NIT）和羟氯胺酮（OCZ） 4种市售片吸虫药的杀灭效果。实验1通过连续给药两剂来评估对TCBZ、RAF和NIT的可能耐药性。实验二通过单剂量TCBZ治疗对RAF和NIT耐药的动物，研究潜在的多药耐药。实验三考察了OCZ作为一种替代疗法的疗效。所有试验均采用粪卵计数减少试验（FECRT）作为主要评估方法。在实验1中，用RAF或NIT处理的牛，即使在第二次剂量后，卵子数量也没有减少。TCBZ处理导致鸡蛋数量在处理后第7天显著下降，但从第14天开始逐渐增加，到第56天恢复到处理前水平。在实验二中，经TCBZ处理的动物在第7天再次显示卵子数量显著下降，随后在第28天稳定上升到基线值。在实验三中，OCZ在第7天完全消除了卵子，有效率为100%。本研究首次提供了埃及动物使用三氯咪唑（TCBZ）治疗失败的文献证据，强调了在全国范围内开展大规模调查的迫切需要。相比之下，羟氯胺（oxyclozanide， OCZ）在消除牛片形虫感染方面表现出高效率，强调了其作为改善埃及片形虫病控制和管理战略的一种有价值的替代方案的潜力。

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引用次数: 0

Genome-wide screening, identification and analysis of BAR domain-containing proteins in Toxoplasma gondii 刚地弓形虫BAR结构域蛋白的全基因组筛选、鉴定和分析。

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-11-19 DOI: 10.1016/j.exppara.2025.109068

Weiwei Sun, An Yan, Lifang Wang, Bohan Wang, Baoliang Pan

Toxoplasma gondii is an important zoonotic pathogen that infects nucleated cells in all warm-blooded animals, and affects about one-third of the world's population. The treatment for T. gondii relies on drugs, but there is no specific cure. The discovery, identification and understanding of key protein families of T. gondii are of great significance for candidate antigen screening, vaccine development, and novel prevention and control program. The Bin, amphiphysin and Rvs (BAR) superfamily is a category of proteins with the BAR domain, which plays an important role in membrane tubulation and constriction during vesicle formation in mammalian cells, and is essential for material transport. However, the information of BAR proteins in T. gondii is not comprehensive. In present study, we screened and identified four BAR domain-containing proteins (Bcps), including TGGT1_259720, TGGT1_320760, TGGT1_232180 and TGGT1_224070 in the T. gondii genome using HMM search and local blast. All above-mentioned Bcps contained the BAR domain, and the 3D structures present the typical crescent shaped helical dimer of BAR. The classification of BAR domains was analyzed by evolutionary tree clustering, and it was found that TGGT1_259720 belonged to F-BAR and the other three belonged to N-BAR. The phylogenetic analysis of four protozoa showed that the Bcps of T. gondii were the closest relative to those of N. caninum, which was consistent with the result of collinearity analysis among species. Moreover, conserved motif and gene composition analysis further confirmed that these closely related proteins are more similar in these structures. The prediction of interacting proteins also showed that Bcps had strong interaction with the key proteins of vesicle transport. The quantification of BAR genes by qPCR showed that these BAR genes were expressed during the growth and proliferation of T. gondii, and the expression patterns were different under different nutritional conditions with the increase of FBS concentration gradient, indicating that these genes played different roles.

刚地弓形虫是一种重要的人畜共患病原体，感染所有温血动物的有核细胞，影响世界上约三分之一的人口。弓形虫的治疗依赖于药物，但没有特效药。弓形虫关键蛋白家族的发现、鉴定和认识，对候选抗原筛选、疫苗研制和新型防控规划具有重要意义。Bin, amphiphysin and Rvs （BAR）超家族是一类具有BAR结构域的蛋白质，在哺乳动物细胞囊泡形成过程中，在膜管和收缩中起重要作用，对物质运输至关重要。然而，关于弓形虫中BAR蛋白的信息并不全面。本研究利用隐马尔可夫搜索和局部blast技术，从弓形虫基因组中筛选出4个BAR结构域蛋白（bps），分别为TGGT1_259720、TGGT1_320760、TGGT1_232180和TGGT1_224070。上述Bcps均含有BAR结构域，三维结构呈现典型的BAR新月形螺旋二聚体。采用进化树聚类方法对BAR结构域进行分类，发现TGGT1_259720属于F-BAR，其余3个属于N-BAR。4种原生动物的系统发育分析表明，弓形虫的Bcps与犬奈虫的Bcps最接近，这与种间共线性分析结果一致。此外，保守基序和基因组成分析进一步证实了这些密切相关的蛋白在这些结构上更加相似。相互作用蛋白的预测也表明，Bcps与囊泡运输的关键蛋白具有较强的相互作用。通过qPCR对BAR基因的定量分析发现，这些BAR基因在弓形虫生长和增殖过程中均有表达，且随着FBS浓度梯度的增加，不同营养条件下BAR基因的表达模式不同，说明这些基因发挥了不同的作用。

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引用次数: 0

Novel anticoagulant proteins identified in the salivary glands of Rhipicephalus sanguineus and their interaction with coagulation factor X 在血鼻猪唾液腺中鉴定的新型抗凝蛋白及其与凝血因子X的相互作用

IF 1.6 4区医学 Q3 PARASITOLOGY

Experimental parasitology

Pub Date : 2025-11-13 DOI: 10.1016/j.exppara.2025.109063

YaJun Lu , YaE Zhao , XingZhi Feng

In the salivary glands of the hematophagous arthropod R. sanguineus, a total of 122 protein coding sequences interacting with coagulation factor X (FX) were identified using the yeast two-hybrid (Y2H) technology, including 100 characterized proteins, and 22 uncharacterized. Three proteins, ferritin-like, tyrosine-protein phosphatase Lar-like, and elongation factor 1 alpha, were selected for cloning, demonstrated significant impacts on four key coagulation parameters: prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), and fibrinogen (FIB) levels. They significantly prolonged PT, APTT, and TT, and reduced FIB levels, indicating interactions with both extrinsic and endogenous coagulation pathways and potential interference with fibrinogen synthesis, metabolism, or functional activity. Docking simulations between the proteins and FX provided evidence of stable, specific protein-protein complexes, mediated by hydrophobic forces. The high confidence scores and diverse interaction modes suggested evolutionary refinement for tight, specific binding to FX. The study introduced promising anticoagulant molecules from hematophagous arthropods, rigorously validated their activity, and highlighted their potential for research and therapeutic applications. These findings have implications for the development of new anticoagulant and antithrombotic therapeutics, underscoring the importance of exploring natural sources for new drugs to expand the drug pipeline in cardiovascular medicine.

利用酵母双杂交（Y2H）技术，在嗜血节肢动物R. sanguineus唾液腺中共鉴定出122个与凝血因子X （FX）相互作用的蛋白编码序列，其中已鉴定蛋白100个，未鉴定蛋白22个。选择铁蛋白样、酪氨酸蛋白磷酸酶样和延伸因子1 α 3种蛋白进行克隆，发现它们对凝血酶原时间（PT）、活化的部分凝血活酶时间（APTT）、凝血酶时间（TT）和纤维蛋白原（FIB）水平等4个关键凝血参数有显著影响。它们显著延长了PT、APTT和TT，并降低了FIB水平，表明它们与外源性和内源性凝血途径相互作用，并可能干扰纤维蛋白原合成、代谢或功能活性。蛋白质和FX之间的对接模拟提供了由疏水力介导的稳定、特异性蛋白质复合物的证据。高置信度分数和多样化的相互作用模式表明，进化改进了与FX的紧密、特异性结合。本研究从食血节肢动物中引入了有前途的抗凝血分子，严格验证了它们的活性，并强调了它们在研究和治疗应用方面的潜力。这些发现对开发新的抗凝血和抗血栓治疗药物具有重要意义，强调了探索新药天然来源以扩大心血管医学药物管道的重要性。

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引用次数: 0