Horses as Sentinels for the Circulation of Flaviviruses in Eastern-Central Germany.

Since 2018, autochthonous West Nile virus (WNV) infections have been regularly reported in eastern-central Germany. While clinically apparent infections in humans and horses are not frequent, seroprevalence studies in horses may allow the tracing of WNV and related flaviviruses transmission, such as tick-borne encephalitis virus (TBEV) and Usutu virus (USUV), and consequently help to estimate the risk of human infections. Hence, the aim of our study was to follow the seropositive ratio against these three viruses in horses in Saxony, Saxony Anhalt, and Brandenburg and to describe their geographic distribution for the year 2021. In early 2022, i.e., before the virus transmission season, sera from 1232 unvaccinated horses were tested using a competitive pan-flavivirus ELISA (cELISA). In order to estimate the true seropositive ratio of infection with WNV, TBEV, and USUV for 2021, positive and equivocal results were confirmed by a virus neutralization test (VNT). In addition, possible risk factors for seropositivity using questionnaires were analyzed using logistic regression based on questionnaires similar to our previous study from 2020. In total, 125 horse sera reacted positive in the cELISA. Based on the VNT, 40 sera showed neutralizing antibodies against WNV, 69 against TBEV, and 5 against USUV. Three sera showed antibodies against more than one virus, and eight were negative based on the VNT. The overall seropositive ratio was 3.3% (95% CI: 2.38-4.40) for WNV, 5.6% (95% CI: 4.44-7.04) for TBEV, and 0.4% (95% CI: 0.14-0.98) for USUV infections. While age and number of horses on the holding were factors predicting TBEV seropositivity, no risk factors were discovered for WNV seropositivity. We conclude that horses are useful sentinels to determine the flavivirus circulation in eastern-central Germany, as long as they are not vaccinated against WNV.

Seroprevalence and Risk Factors for Equine West Nile Virus Infections in Eastern Germany, 2020.

West Nile virus (WNV) infections were first detected in Germany in 2018, but information about WNV seroprevalence in horses is limited. The study's overall goal was to gather information that would help veterinarians, horse owners, and veterinary-, and public health- authorities understand the spread of WNV in Germany and direct protective measures. For this purpose, WNV seroprevalence was determined in counties with and without previously registered WNV infections in horses, and risk factors for seropositivity were estimated. The cohort consisted of privately owned horses from nine counties in Eastern Germany. A total of 940 serum samples was tested by competitive panflavivirus ELISA (cELISA), and reactive samples were further tested by WNV IgM capture ELISA and confirmed by virus neutralization test (VNT). Information about potential risk factors was recorded by questionnaire and analyzed by logistic regression. A total of 106 serum samples showed antibodies against flaviviruses by cELISA, of which six tested positive for WNV IgM. The VNT verified a WNV infection for 54 samples (50.9%), while 35 sera neutralized tick-borne encephalitis virus (33.0%), and eight sera neutralized Usutu virus (7.5%). Hence, seroprevalence for WNV infection was 5.8% on average and was significantly higher in counties with previously registered infections (p = 0.005). The risk factor analysis showed breed type (pony), housing in counties with previously registered infections, housing type (24 h turn-out), and presence of outdoor shelter as the main significant risk factors for seropositivity. In conclusion, we estimated the extent of WNV infection in the resident horse population in Eastern Germany and showed that seroprevalence was higher in counties with previously registered equine WNV infections.

[Assessment of selenium status in relation to the supplementation of selenium enriched mineral licks and mineral feeds in equines in Thuringia]. / Selenstatus in Thüringer Pferdehaltungen in Abhängigkeit des Einsatzes selenhaltiger Mineral- und Ergänzungsfuttermittel.

OBJECTIVE: Screening of commercial mineral feeds and mineral licks on the German market containing selenium (Se) in relation to the Se status in equines in Thuringia with different forms of Se supplementation. MATERIAL AND METHODS: Commercially available Se-containing minerals for horses identified by an online research were evaluated for their Se concentration, Se source, flavour carrier and recommended dosage according to the manufacturer's labelling. Furthermore, serum Se status in 8 equine farms was regularly monitored over the period of one year. The sampled horses either received no Se supplement or Se was supplemented by a mineral lick, a mineral feed or supplementary feed. RESULTS: In 29.7 % of the mineral licks, the manufacturers provided a label with information on the maximal daily Se dosage, all of which exceeded the current recommendations of daily Se intake for a 600 kg horse under maintenance conditions. According to the manufacturers' labelled dosage, 67.5 % of the mineral feeds also exceeded the recommended daily Se intake taking the daily Se requirement of a 600 kg horse under maintenance conditions into consideration. The declarations of the feeds, especially in the case of mineral licks, were frequently not in conformity with the Regulation (EC) Nr. 767/2009 on commercial market introduction and use of feed. Concerning the horses' Se status, only horses receiving Se supplementation either via mineral lick with a Se concentration ≥ 20-50 mg/kg or a mineral or supplementary feed exhibited median serum Se concentrations within the reference range. In 2 of 10 horses with access to mineral licks with molasses containing Se concentrations ≥ 45 mg/kg, serum Se concentration exceeded the reference range at at least at one sampling time point. CONCLUSION AND CLINICAL RELEVANCE: Due to difficulties in their dosaging, the use of mineral licks should be only considered in extensively managed farms with limited access to the individual horse. The recommended Se concentration in mineral licks is recommended to lie in the range of 40-50 mg/kg. In order to attain improved control over Se intake, this trace element ought to be provided by individual feeding using either a mineral supplement or a supplementary feed.

[Paratuberculosis in a miniature donkey (Equus asinus f. asinus)]. / Paratuberkulose bei einem Zwergesel (Equus asinus f. asinus).

Paratuberculosis is mainly an infectious disease of ruminants with worldwide distribution. Infection occurs in early stages of life. Other animal species beyond ruminants are rarely affected, however, experimental and natural infections are possible. A case of paratuberculosis in a miniature donkey (Equus asinus f. asinus) with typical clinical and pathomorphological changes is reported here. Lesions were mainly observed in the intestine. Causative for the profuse diarrhoea with emaciation was massive diffuse granulomatous enteritis involving large quantities of acid-fast organism mainly in macrophages. Granulomatous inflammation with acid-fast bacilli again in macrophages to a lesser degree could be detected in the liver. Mycobacterium avium subsp. paratuberculosis (MAP) was isolated from intestinal contents after an incubation period of four weeks. MAP-specific DNA (IS900 and f57) was detected by polymerase chain reaction in culture material. Additionally MAP-isolates were characterized by multi-target genotyping (MIRU-VNTR- and MLSSR-typing). Isolates belonged to the Type II group and exhibited a unique genotype different from other MAP strains in Germany. The donkey originated from a donkey breeding farm in France with intensive free ranging cattle in the neighbourhood and could have been infected there. Donkeys should be considered as paratuberculosis-susceptible animals in exceptional cases and as possible reservoirs or disseminators of infection.