Development of a multiplex PCR for identification of Dictyocaulus lungworms in domestic and wild ruminants

Abstract

Dictyocaulus lungworms are the causative agents of parasitic bronchitis (dictyocaulosis) characterised by coughing and severe lung pathology in domestic and wild ruminants. The objective of this study was to design a simple molecular test that could detect of lungworm DNA from both adult and larval lungworms and could distinguish between the most common Dictyocaulus species found in cattle and in some species of wild ruminants. A multiplex PCR test with four novel primers targeting species-specific regions of the second internal transcribed spacer (ITS2) was designed based on our own sequence data as well as on available sequence information in GenBank. After PCR amplification of lungworms from European bison (Bison bonasus), cattle (Bos taurus), moose (Alces alces), red deer (Cervus elaphus) and roe deer (Capreolus capreolus), products were analysed with gel electrophoresis. This resulted in three specific bands of different size depending on the species analysed. Dictyocaulus viviparus collected from cattle or European bison resulted in a ca. 560 bp band, D. capreolus collected from roe deer produced a band ca. 400 bp and the longest DNA band (ca. 660 bp) was obtained with DNA from Dictyocaulus sp. collected from red deer and moose. Dictyocaulus eckerti bands with expected size of 714 bp were not observed in our study. The multiplex method produced consistent results with samples from both Sweden and Poland and overcame the limitations of traditional techniques based on differences in morphological features of parasites at different life stages.

Dictiocaulose Em Bovinos No Município De Londrina, Pr, Brasil

“…Recentemente, devido à importância econômica da doença, uma técnica de diagnóstico molecular baseada em PCR multiplex foi desenvolvida, permitindo o diagnóstico e a identificação precisa de Dictyocaulus spp. em ruminantes domésticos e silvestres (11).…”

Severity and prevalence of small lungworm infection on three South Australian farms and associations with sheep carcass characteristics

“…Additionally, the study found no association between the presence of small lungworm lesions and pneumonia. It also highlighted the limitations of currently available diagnostic tests for small lungworm on farm and for assessing infections at the processing speeds of a commercial abattoir (McRae et al, 2016;Pyziel et al, 2015;Vina et al, 2013). A previous unpublished study in southeast SA had suggested small lungworm lesions reduced carcass weight (Webb Ware unpublished data).…”

“…However, this method was not validated, cannot be performed at abattoir processing speed, does not allow for diagnosis of the species present and must occur post-mortem. Alternatively, a genus-or species-specific molecular test could be developed for sheep lungworms (Pyziel et al, 2015), which would also allow for improved testing whilst sheep were on the farm. It would also provide a better indication of the effect of a current patent infection on liveweight as patent infections may have a more significant effect on sheep productivity (Sauerlander, 1988).…”

Molecular characterization of Dictyocaulus nematodes in wild red deer Cervus elaphus in two areas of the Italian Alps

“…In free-living cervids, the disease should be considered noteworthy for wildlife management with respect to the interaction with livestock (Pyziel et al 2015 ), as well as to potential threats to biodiversity, wildlife conservation, and the development of the game industry (Pyziel et al 2017 ; Pyziel 2018 ). Some species of the genus Dictyocaulus show host-specificity, as in the case of Dictyocaulus viviparus Bloch, 1782 (cattle) and Dictyocaulus filaria Rudolphi, 1809 (sheep and goat) (Bangoura et al 2020 ).…”