Trichinella spp. in Wild Boars (Sus scrofa), Brown Bears (Ursus arctos), Eurasian Lynxes (Lynx lynx) and Badgers (Meles meles) in Estonia, 2007-2014

Simple Summary Trichinellosis is an important foodborne zoonosis. In Estonia, Trichinella infections are endemic in wild animals. This paper summarizes findings of Trichinella-parasites during an 8-year period in Estonia in selected host species: wild boars, brown bears, Eurasian lynxes, and badgers. The results highlight that testing wildlife hunted for human consumption for Trichinella is important, and that there is room for improvement in the proportion of hunted animals tested. In this study, we summarize Trichinella findings from four wild, free-ranging host species from Estonia during 2007-2014. Trichinella spp. larvae were detected in 281 (0.9%, 95% confidence interval (CI) 0.8-1.0) of 30,566 wild boars (Sus scrofa), 63 (14.7%, 95% CI 11.6-18.3) of 429 brown bears (Ursus arctos), 59 (65.56%, 95% CI 55.3-74.8) of 90 Eurasian lynxes (Lynx lynx), and three (60.0%, 95% CI 18.2-92.7) of five badgers (Meles meles). All four European Trichinella species were detected: T. britovi in 0.7% of the wild boars, 7.2% of the brown bears, 45.6% of the lynxes, and 40.0% of the badgers; T. nativa in 0.1% of the wild boars, 5.8% of the brown bears, and 20.0% of the lynxes; T. pseudospiralis in 0.02% the wild boars; and T. spiralis in 0.03% of the wild boars and 4.4% of the lynxes. The results include the first description from Estonia of T. britovi in brown bear and badgers, T. pseudospiralis in wild boars, and T. spiralis in wild boars and lynxes. The results indicate high infection pressure in the sylvatic cycles across the years-illustrating continuous risk of spillover to domestic cycles and of transmission to humans.Peer reviewe

Trichinella spp. biomass has increased in raccoon dogs (Nyctereutes procyonoides) and red foxes (Vulpes vulpes) in Estonia

Background: Raccoon dogs and red foxes are well-adapted hosts for Trichinella spp. The aims of this study were to estimate Trichinella infection prevalence and biomass and to investigate which Trichinella species circulated in these indicator hosts in Estonia. Methods: From material collected for evaluating the effectiveness of oral vaccination program for rabies eradication in wildlife, samples from 113 raccoon dogs and 87 red foxes were included in this study. From each animal, 20 g of masseter muscle tissue was tested for the presence of Trichinella larvae using an artificial digestion method. The Trichinella larvae were identified to species level by multiplex polymerase chain reaction method. Results: The majority of tested animals were infected with Trichinella spp. The parasite species identified were T. nativa and T. britovi. The apparent infection prevalence was 57.5% in raccoon dogs and 69.0% in red foxes, which were higher than previous estimates. In addition, the larval burden had also increased in both hosts. We estimated that in 2011-2012, the Trichinella spp. biomass was more than 15 times higher in raccoon dogs and almost two times higher in red foxes than in 1992-2000 (based on mean larval burden), and almost 20 times higher in raccoon dogs and almost five times higher in red foxes than in 2000-2002 (based on median larval burden). Conclusions: Raccoon dogs and red foxes are relevant reservoirs for Trichinella spp. in Estonia. The biomass of Trichinella circulating in sylvatic cycles was substantial and had increased: there is substantial infection pressure in the sylvatic cycle.Peer reviewe

Occurence of Eserichia coli O157:H7 in Estonian dairy farms and beef production chain in 2005–2014

Escherichia coli O157:H7 human infections are mostly associated with consumption of raw or undercooked beef and raw milk. Ruminants, especially cattle are the main reservoir of the pathogen. The main aim of the present study was to evaluate the occurrence of E. coli O157:H7 in cattle dairy farm and beef production level in Estonia. It was found that E. coli O157:H7 is present at both the dairy farm and slaughterhouse level. The occurrence of the pathogen at Estonian dairy farm level was 1.2% from 1312 cattle's sampled. At slaughterhouse level 744 hide swabs were taken from which 30 (4.0%) were positive to E. coli O157:H7. Both stx1 and stx2 genes were determined from isolates and often the combination of both genes was found to be present. Minimal inhibitory concentration was determined for 30 E. coli O157:H7 isolates which revealed 26.7% of multiresistant isolates. In 2009–2010 in a total of 188 beef samples were analysed, which all were negative for O157:H7

Molecular Characterization of African Swine Fever Virus Isolates in Estonia in 2014–2019

After the extensive spread of the African swine fever virus (ASFV) genotype II in Eastern Europe, the first case of African swine fever (ASF) in Estonia was diagnosed in September 2014. By the end of 2019, 3971 ASFV-positive wild boars were found, and 27 domestic pig outbreaks were reported. A selection of ASFV isolates from wild boar and domestic pigs (during the period of September 2014–2019) was molecularly characterized using standardized genotyping procedures. One of the proven markers to characterize this virus is the central variable region (CVR) within the B602L gene. In summer 2015, a new ASFV genotype II CVR variant 2 (GII-CVR2) was confirmed in Estonia. The results suggest that the GII-CVR2 variant was only confirmed in wild boar from a limited area in southern Estonia in 2015 and 2016. In addition to GII-CVR2, a single nucleotide polymorphism (SNP) that resulted in amino acid change was identified within the genotype II CVR variant 1 (GII-CVR1). The GII-CVR1/SNP1 strain was isolated in Estonia in November 2016. Additional GII-CVR1/SNP1 cases were confirmed in two neighbouring counties, as well as in one outbreak farm in June 2017. Based on the available data, no GII-CVR2 and GII-CVR1/SNP1 have been reported by other affected European countries. The spread of variant strains in Estonia has been limited over time, and restricted to a relatively small area

Wild boar (Sus scrofa) carcasses as an attraction for scavengers and a potential source for soil contamination with the African swine fever virus

The wild boar (Sus scrofa) is a social animal species native to Eurasia. During the last decade, the wild boar population in Estonia has been severely affected by the African swine fever virus (ASFV), which has also affected domestic pig farming. The potential transmission routes of ASFV remain unclear and are currently under intensive investigation. This pilot study aimed to clarify the frequency and characteristics of contacts between living wild boars and the carcasses of their conspecifics, which could play a role in the transmission of ASFV. Wild animals' contact and scavenging behavior on wild boar carcasses were studied using trail cameras in an experimental setting on Hiiumaa, Western Estonia. Four legally hunted carcasses were used in the present study. This study aimed to determine whether intraspecies scavenging occurs in wild boars. The persistence of ASFV DNA in soil contaminated with infected wild boar carcasses was investigated separately. Among the 17 identified wildlife species that visited wild boar carcasses, the common raven (Corvus corax) was the most frequent one (37.26%), followed by raccoon dogs (Nyctereutes procyonoides; 4.25%), carcass conspecific/wild boars (3.16%), and red foxes (Vulpes vulpes; 2.14%). Regarding the direct contact with the carcass, the same species ranking was detected: common raven (74.95%), raccoon dogs (9.94%), wild boars (4.21%), and red foxes (4.21%). No clear signs of cannibalism were noted among the wild boars, although brief physical contact with the carcasses was evident. The persistence of ASFV DNA in soil contaminated by infected wild boar carcasses was investigated separately. This study revealed that ASFV DNA from infected carcasses could be detected in forest soil for prolonged periods, even after removing the carcasses. Hence, the carcasses of infected wild boars may play an important role in spreading the African swine fever virus in wild boar populations; thus, prompt removal and disinfection of the soil could be considered necessary to limit the spread of the infection