Ciuca L, Vismarra A, Lebon W, Beugnet F, Morchon R, Rinaldi L, Cringoli G, Kramer L, Genchi M
Ion Ionescu de la Brad, University of Agricultural Sciences and Veterinary Medicine Iasi, Faculty of Veterinary Medicine, M. Sadoveanu Alley No. 8, 700489, Iasi, Romania; Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via della Veterinaria, 1, Naples, Italy.
Dipartimento di Scienze Medico-Veterinarie, Università di Parma, via del Taglio 10, 43126, Parma, Italy.
Vet Parasitol. 2020;277S:100029. doi: 10.1016/j.vpoa.2020.100029. Epub 2020 Aug 5.
Dogs are the primary host for Dirofilaria repens, therefore it is mandatory to accurately diagnose the canine infection and to expand our current knowledge on parasite biology and the immune response of the infected host for a better prevention.Thus, the aim of the present study was to provide new insights from experimental infections of dogs with D. repens, focusing on the evaluation of: 1) the pre-patent period and 2) the antibody response against D. repens somatic antigens and against the Wolbachia endosymbiont. Briefly, on Day 0, twenty purpose-bred Beagle dogs were experimentally infected with 50 infective larvae (L3) of D. repens. Starting from Day 58 until the last day of the study (Day 281), blood samples were collected on a monthly basis for detection of antibodies against D. repens (Dr) and recombinant Wolbachia surface protein (rWSP) by non-commercial IgG-ELISAs. Additional samples were collected on Days 220, 245 and 281 for the detection of microfilariae (mff) using the modified Knott's test and biomolecular analysis, following two PCR protocols: Gioia et al. (2010; protocol A) and Rishniw et al. (2006- protocol B). The results were analysed by univariate statistical analyses using 2×2 contingency tables and K Cohen was calculated to assess the agreement among all the diagnostic techniques. Overall, the outcome of the study revealed that out of the 20 dogs experimentally infected with D. repens, 16 (80 %) were microfilaraemic, 17 (85 %) were positive at DNA detection in the blood, 18 (90 %) had D. repens antibodies and 16 (80 %) had Wolbachia antibodies on the last day of the study. The overall k agreement between Knott's and PCR protocol B was 0.442 (P=0.0001) and increased throughout the study, reaching 0.828 (P=0.0001) on Day 281. To the authors knowledge, this is only the second study reporting antibody response to D. repens somatic antigen in experimentally infected dogs. ELISA results showed that an antibody response develops before the onset of patency, and steadily increases with time. Results would suggest that the development of an immunological response to infection could lead to application in epidemiological studies, risk assessment and as an aid in the diagnostic approach in dogs, in particular for early infections without mff.
犬是匐行恶丝虫的主要宿主，因此准确诊断犬感染情况并拓展我们目前对寄生虫生物学及感染宿主免疫反应的认识对于更好地预防感染至关重要。因此，本研究的目的是通过对犬进行匐行恶丝虫实验性感染来提供新的见解，重点评估：1）潜隐期；2）针对匐行恶丝虫体抗原和沃尔巴克氏体共生菌的抗体反应。简而言之，在第0天，20只经专门培育的比格犬被实验性感染50条匐行恶丝虫感染性幼虫（L3）。从第58天开始直至研究的最后一天（第281天），每月采集血样，通过非商业IgG-ELISA检测针对匐行恶丝虫（Dr）和重组沃尔巴克氏体表面蛋白（rWSP）的抗体。在第220天、245天和281天采集额外样本，使用改良的Knott氏试验检测微丝蚴（mff），并按照两种PCR方案进行生物分子分析：Gioia等人（2010年；方案A）和Rishniw等人（2006年 - 方案B）。使用2×2列联表通过单变量统计分析对结果进行分析，并计算K Cohen值以评估所有诊断技术之间的一致性。总体而言，研究结果显示，在20只经实验感染匐行恶丝虫的犬中，16只（80%）出现微丝蚴血症，17只（85%）血液DNA检测呈阳性，18只（90%）有匐行恶丝虫抗体，16只（80%）在研究最后一天有沃尔巴克氏体抗体。Knott氏试验与PCR方案B之间的总体k一致性为0.442（P = 0.0001），且在整个研究过程中有所增加，在第281天达到0.828（P = 0.0001）。据作者所知，这是第二项报道实验感染犬对匐行恶丝虫体抗原抗体反应的研究。ELISA结果表明，在虫体出现之前抗体反应就已产生，并随时间稳步增加。结果表明，针对感染的免疫反应的发展可能会应用于流行病学研究、风险评估，并有助于犬的诊断方法，特别是对于无微丝蚴的早期感染。
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