Hepatitis E virus (HEV) is one of the major causes of acute viral hepatitis worldwide. In Europe, food-borne zoonotic transmission of HEV genotype 3 has been associated with domestic pigs and wild boar. Controversial data are available on the circulation of the virus in animals that are used for human consumption, and to date, no gold standard has yet been defined for the diagnosis of HEV-associated hepatitis. To investigate the current HEV infection status in Lithuanian pigs and wild ungulates, the presence of viral RNA was analyzed by nested reverse transcription polymerase chain reaction (RT-nPCR) in randomly selected samples, and the viral RNA was subsequently genotyped. In total, 32.98 and 22.55% of the domestic pig samples were HEV-positive using RT-nPCR targeting the ORF1 and ORF2 fragments, respectively. Among ungulates, 25.94% of the wild boar samples, 22.58% of the roe deer samples, 6.67% of the red deer samples and 7.69% of the moose samples were positive for HEV RNA using primers targeting the ORF1 fragment. Using primers targeting the ORF2 fragment of the HEV genome, viral RNA was only detected in 17.03% of the wild boar samples and 12.90% of the roe deer samples. Phylogenetic analysis based on a 348-nucleotide-long region of the HEV ORF2 showed that all obtained sequences detected in Lithuanian domestic pigs and wildlife belonged to genotype 3. In this study, the sequences identified from pigs, wild boars and roe deer clustered within the 3i subtype reference sequences from the GenBank database. The sequences obtained from pig farms located in two different counties of Lithuania were of the HEV 3f subtype. The wild boar sequences clustered within subtypes 3i and 3h, clearly indicating that wild boars can harbor additional subtypes of HEV. For the first time, the ORF2 nucleotide sequences obtained from roe deer proved that HEV subtype 3i can be found in a novel host. The results of the viral prevalence and phylogenetic analyses clearly demonstrated viral infection in Lithuanian pigs and wild ungulates, thus highlighting a significant concern for zoonotic virus transmission through both the food chain and direct contact with animals. Unexpected HEV genotype 3 subtype diversity in Lithuania and neighboring countries revealed that further studies are necessary to understand the mode of HEV transmission between animals and humans in the Baltic States region.

Hepatitis E is an important public health concern disease that causes acute hepatitis in humans with different hepatitis E virus strains reported among domestic and wild animals. In Lithuania, seroprevalence studies on animals considered to be possible virus reservoirs, particularly in wildlife species, had never been investigated before. In order to assess the actual distribution of the virus in the domestic pig and wildlife species, serum samples originating from different geographic areas of Lithuania were tested for the presence of antibodies to the virus using enzyme-linked immunosorbent assay. The study results proved that hepatitis E virus infection is prevalent among domestic pigs, wild boar, moose, and roe deer in Lithuania, with the overall prevalence of IgG antibodies against hepatitis E virus being 43.75%, 57.05%, 11.76%, and 1.20%, respectively. Significantly higher ( P < 0.01) seroprevalence values of 53.66% and 80% were observed in weaned pigs and adult wild boar. Herds with 15 001–30 000 pigs were 2.4 times more likely ( P < 0.01) to test positive for hepatitis E virus antibodies (70.18%), than herds with ≤ 500 pigs (21.11%). The differences in seroprevalence rates between domestic pigs (from 25 to 66.67%) and wild boar (from 42.86 to 73.33% ) in all 10 counties were not significant ( P > 0.05). Estimated low seroprevalence rate in moose, roe deer and absence of antibodies in red deer suggest that such wildlife species are accidental hosts of hepatitis E virus or at least hepatitis E virus is not present in Lithuanian cervides.

Background: Porcine reproductive and respiratory syndrome virus (PRRSV) is recognized worldwide as an important and economically devastating pathogen in pig production. Although PRRSV is widespread in domestic swine, there is a lack of information regarding PRRSV infection in European wild boars ( Sus scrofa ). Currently available information does not provide conclusive evidence that wild boars are a reservoir of PRRSV. Nevertheless, wild boars may be likely to become infected by domestic swine through occasional direct or indirect contact. Furthermore, wild boars can act as a reservoir for infectious diseases of domestic pigs. Therefore, the objectives of the present study were to determine the virus prevalence and further explore the epidemiology and diversity of PRRSV strains present in Lithuanian wild boars over a 5-year period. A total of 1597 tissue and serum samples from wild boars inhabiting 44 districts and ten counties in Lithuania were analysed using conventional nested reverse transcription polymerase chain reaction (RT-PCR) and real-time Taqman RT-PCR for the detection of PRRSV-specific open reading frame (ORF) 1 and 6 sequences. Results: PRRSV was highly prevalent in Lithuanian wild boar populations, with an average rate of 18.66 % using conventional RT-PCR and 19.54 % using real-time RT-PCR. PRRSV was detected in 36.71 and 41.77 % of 237 hunting grounds tested by conventional RT-nPCR and real-time RT-PCR, respectively. No statistically significant differences in PRRSV prevalence were observed by geographic area in the ten Lithuanian counties. Animals infected with PRRSV were identified in all age groups; however, significantly higher prevalence rates were identified in subadult and adult wild boars than in juveniles up to 12 months old. No positive results were obtained using conventional PCR with Type 2 specific primers. Phylogenetic analysis of the partial ORF5 region revealed that ten wild boars harboured virus sequences belonging to genetic subtypes 3 and 4 and may therefore pose a serious threat to Lithuanian pig farms in which only subtype two strains are circulating. Conclusions: The results of virus prevalence and phylogenetic analyses strongly support the role of wild boars as a possible natural reservoir for PRRSV in Lithuania.

There is an increased interest in using automatic milking systems (AMS) to indirectly assess the welfare of dairy cows, but knowledge on analyzing the association between lameness, milk yield characteristics, and reproductive performance in cows is still insufficient. The main aims of this study were to evaluate the influence of lameness on several AMS variables and reproductive performance indicators during the early stage of lactation and estrus in Lithuanian Black and White dairy cows, as well as to assess the associations between lameness, productivity and reproductive efficiency. A total of 418 milking cows (50.3 +/- 1.2 d postpartum) without any apparent reproductive disorder were monitored for hoof health status. Cows were assigned to two groups on the basis of visual locomotion scoring: "non-lame"cows (group 1; 74.20%) and cows presenting "lameness" (lame cows) (group 2; 25.80%). Productive and milking performances of dairy cows were recorded from 50 to 100 days in milk (DIM) and 1 day after the first estrus. The lameness was predominantly localized on the hind feet (79.60%) and less frequently - on the front feet (20.40%; p<0.001). Furthermore, the lameness had a tendency to decrease milk production (4.24%; p<0.05) and increase the difference in milk yield between rear and front quarters of the udder (1.20%; p<0.05). The frequency of milking (5.19%) was lower in lame cows (p<0.05). The lame cows during estrus showed a more pronounced decrement in milk yield and milking frequency (p<0.05), and also higher milk progesterone concentration values (1.55-1.76 time's; p<0.001), and an increasing number of inseminations (11.69%; p<0.05) were observed. The results highlighted that analysis of data from AMS programs can be a successful tool for reducing risk factors related to the effective management of reproductive performance and hoof health of dairy cows.

Following publication of the original article [1], we have been notified of a typing error in the “Discussion” section.

This study evaluated infrared thermography (IRT) as a dual-purpose, non-invasive tool for detecting limb pathology and subclinical mastitis in dairy cows. Surface temperatures were measured at the coffin, pastern, and fetlock regions of all limbs and at the teat surface of all udder quarters. Limb health status was determined by orthopedic clinical examination, and mastitis status was assessed using the California Mastitis Test (CMT). Hindlimbs were significantly warmer than forelimbs in healthy cows (p < 0.001), with confirmed bilateral symmetry (r = 0.524–0.569). Limbs with pathology showed significantly higher temperatures at all anatomical sites, with particularly large effect sizes in the hindlimbs (r around 0.80). Additional lesion-specific analyses demonstrated that both infectious and claw horn lesions exhibited significantly higher temperatures than healthy limbs. In the hindlimbs, infectious lesions showed higher thermographic values than claw horn lesions. CMT-positive quarters showed teat temperatures of 30.0–30.2 °C compared with 25.2–25.8 °C in CMT-negative quarters (p < 0.001). Receiver operating characteristic (ROC) analysis demonstrated excellent diagnostic performance for teat surface temperature in identifying CMT-positive quarters, with an area under the ROC curve (AUC) of 0.956. The optimal threshold was 29.5 °C, corresponding to a sensitivity of 0.992 and a specificity of 0.838. Each 1 °C increase in teat temperature increased the odds of CMT positivity by 5.3-fold (p < 0.001). A practical three-zone thermographic framework was established: <27 °C (healthy), 27–29.5 °C (monitoring recommended), and ≥29.5 °C (mastitis likely). Teat temperature was not associated with composite milk somatic cell count, but hindlimb pathology was independently associated with elevated teat temperatures (partial r = 0.23–0.26, p ≤ 0.001). These findings demonstrate that IRT has strong potential as an integrated screening tool for dairy cow limb health and udder health monitoring.