Veterinary Research Forum

Abstract Trichinella britovi is a species circulating in Iran and has already been observed in wild and domestic carnivores. The present study aims to determine the occurrence of Trichinella species in Iran's most important provinces, including the northern, northeastern, and northwest regions. In total, 93 carcasses of golden jackals, collected due to road accidents between 2019 and 2022, have been obtained from Ardabil, Mazandaran, Golestan, North Khorasan, and Khorasan Razavi provinces, Iran. Genomic DNA (four positives out of 93 carcasses) from each Trichinella genotype was amplified in a multiplex polymerase chain reaction (PCR) reaction. To confirm the multiplex PCR results, the sequences of the PCR products with cytochrome oxidase subunit I were sent to a reference laboratory within the European Union. Bioinformatics software was used to verify the sequencing results of primer fragments and to compare and analyze the results obtained with the sequences stored in the NCBI genetic database. Following multiplex PCR amplification, a two-band pattern was observed. Trichinella britovi larvae displayed the expected band pattern of 127 bp and 253 bp. The PCR products amplified using primers specific for the cytochrome oxidase subunit I gene showed 861 base pair bands in all four samples analyzed. The obtained sequences of cytochrome oxidase subunit I from all four individual muscle larvae had 100% identical haplotypes except for one nucleotide transversion in one sequence. As a result of this study, valuable insights into T. britovi prevalence and phylogenetic distribution among Iranian golden jackals have been gained in terms of prevalence and phylogenic distribution.

Abstract Rhipicephalus (Boophilus) spp. are important vectors for Babesia and Anaplasma species causing severe economic losses in livestock. Chemical compounds are commonly used to control tick infestation; however, acaricides resistance in tick has led to move toward alternative strategies such as vaccination. In this study, we introduced a vaccine candidate, namely CaTro against Rh. microplus tick composing of immunogenic B-cell epitopes derived from Rh. microplus cathepsin L and tropomyosin proteins. To evaluate this vaccine candidate, firstly the CaTro sequence was inserted into the prokaryotic expression vector and the recombinant protein CaTro was expressed in Bl21 bacteria. Afterward, purification was performed by Ni-NTA affinity chromatography. The quality of purified recombinant CaTro was also analyzed using sodium dodecyl sulfate-gel electrophoresis and western blotting. Moreover, to evaluate the induction of immune response, the rabbits were immunized with purified recombinant protein combined with Freund’s adjuvant. The findings of this study revealed molecular weight of expressed protein (CaTro) as 38.00 kDa. Furthermore, anti-CaTro antibody was detected in immunized rabbit's sera through dot blotting; while, there was not any response to the control rabbit's sera. The results suggest that CaTro is a potential candidate to develop an anti- Rh. microplus tick.