Molecular characterization of Echinococcus species in dogs from four regions of Kenya

Prevalence and Genotyping of Echinococcus Species from Livestock in Kajiado County, Kenya

“…The first nested PCR (entire nad1 gene) was performed as described by Hüttner and Nakao [17]. The cyst materials negative using the first PCR assay were genotyped using a second nested PCR as described by Mulinge and Magambo [18], which amplifies part of the nad1 gene (545-552 bp). In both PCR assays, the reaction mixture contained 2 μ l of the DNA, 1 × DreamTaq Green Buffer (20 mM Tris-HCl (pH 8.0), 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, 0.5 % (v/v) Nonidet P40, 0.5 % (v/v) Tween 20) (Thermo Scientific), 0.2 mM dNTPs, 0.25 μ M of forward and reverse primers, 2 mM MgCl 2 , and 0.625 units of DreamTaq Green DNA Polymerase (Thermo Scientific) in 25 μ l final volume.…”

“…Positive PCR products were genotyped by RFLP to the specific Echinococcus species. To this end, 10 μ l of the nested PCR products was digested using 0.5 μ l (5 U) of Hph I restriction enzyme, 1 × Buffer, and 7.5 μ l of nuclease-free water and incubated at 37°C overnight [18, 19]. Positive controls for E. granulosus s. s., E. ortleppi, E. canadensis (G6/7), and E. felidis were resolved alongside the test samples for both methods.…”

The Molecular Epidemiology of Echinococcus Infections

“…Although infections in lions was known to occur for many years, it is only relatively recently that adult worms from lions were characterised using molecular tools and confirmed to be a distinct species, E. felidis [ 31 ]. Interestingly, recent ME studies have demonstrated that E. felidis is not confined to a sylvatic cycle and has been identified in domestic dogs in Kenya [ 32 ]. ME studies have also demonstrated infections with E. granulosus, E. intermedius , and E. ortleppi in dogs, including some mixed infections [ 32 ].…”

“…Interestingly, recent ME studies have demonstrated that E. felidis is not confined to a sylvatic cycle and has been identified in domestic dogs in Kenya [ 32 ]. ME studies have also demonstrated infections with E. granulosus, E. intermedius , and E. ortleppi in dogs, including some mixed infections [ 32 ].…”

“…In endemic regions where multiple species of Echinococcus are maintained in sympatric cycles of transmission, and humans are at risk of infection, it is important to determine the species responsible for clinical infections. This will aid clinical interventions and management [ 32 ] but also inform and guide public health measures. Molecular diagnostic approaches have been shown to be valuable tools in this respect.…”