First detection of Echinococcus multilocularis in Bosnia and Herzegovina

Abstract

Echinococcus multilocularis has been spreading through Central Eastern Europe but has not yet been reported in Bosnia and Herzegovina (B&H). Recently, this parasite is confirmed in Croatia suggesting the movement of the parasite's distribution limit further south. Given that there is no surveillance or monitoring system for echinococcosis in B&H, our study was designed as a pilot study of E. multilocularis. A total of 57 red foxes originating from 24 localities all over the country were collected during the routine rabies monitoring, autopsied and examined for the presence of echinococcosis. Based on intestinal scraping technique and microscopy, E. multilocularis adult worms have been detected in one (1/57, 1.75%) red fox. To verify this finding and to differentiate Echinococcus spp., DNA extracted from adult worms was subjected to species-specific PCR targeting part of the mitochondrial 12S ribosomal RNA gene. E. multilocularis PCR-positive samples were further confirmed by NGS sequencing of a 203 bp amplified fragment of 12S rRNA, which has been deposited in GenBank (Accession no.: OP047920). This finding represents the first detection of E. multilocularis in B&H, strongly suggesting its presence in the country. The confirmation of the parasite in the same locality where migrants/refugees temporarily stay on their route to Western Europe highlights the need for a One Health approach in addressing all future questions. Moreover, the first detection of E. multilocularis in B&H warrants the need for the implementation of an appropriate state surveillance program.

Keywords: Alveolar echinococcosis; Bosnia and Herzegovina; Echinococcus multilocularis; NGS; PCR; Red fox.

Graphical abstract

Fig. 1

Study area localities (dots) and collection sites for red foxes were included in the study. The red dot shows the location with a determined E. multilocularis positive red fox. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 2

The E. multilocularis adult parasite was determined in the small intestines of the infested red fox using the intestinal scraping technique (stereomicroscopic magnification x100). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)