[3 cases of abomasitis in calves associated with Sarcina sp. infection]. / 3 Fälle von Abomasitis im Zusammenhang mit einer Infektion mit Sarcina sp. beim Kalb.

In this case report - to the best knowledge of the authors for the first time in the German-speaking region - 3 calves with ulcerating or emphysematous abomasitis respectively are presented, in which intralesional bacteria of the Sarcina species were identified. The unusual appearance of these bacteria is described, their etiopathogenic relevance discussed.

Serological screening in wild ruminants in Germany, 2021/2022: No evidence of SARS-CoV-2, bluetongue virus or pestivirus spread but high seroprevalences against Schmallenberg virus.

Wildlife animals may be susceptible to multiple infectious agents of public health or veterinary relevance, thereby potentially forming a reservoir that bears the constant risk of re-introduction into the human or livestock population. Here, we serologically investigated 493 wild ruminant samples collected in the 2021/2022 hunting season in Germany for the presence of antibodies against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and four viruses pathogenic to domestic ruminants, namely, the orthobunyavirus Schmallenberg virus (SBV), the reovirus bluetongue virus (BTV) and ruminant pestiviruses like bovine viral diarrhoea virus or border disease virus. The animal species comprised fallow deer, red deer, roe deer, mouflon and wisent. For coronavirus serology, additional 307 fallow, roe and red deer samples collected between 2017 and 2020 at three military training areas were included. While antibodies against SBV could be detected in about 13.6% of the samples collected in 2021/2022, only one fallow deer of unknown age tested positive for anti-BTV antibodies, and all samples reacted negative for antibodies against ruminant pestiviruses. In an ELISA based on the receptor-binding domain (RBD) of SARS-CoV-2, 25 out of 493 (5.1%) samples collected in autumn and winter 2021/2022 scored positive. This sero-reactivity could not be confirmed by the highly specific virus neutralisation test, occurred also in 2017, 2018 and 2019, that is, prior to the human SARS-CoV-2 pandemic, and was likewise observed against the RBD of the related SARS-CoV-1. Therefore, the SARS-CoV-2 sero-reactivity was most likely induced by another hitherto unknown deer virus belonging to the subgenus Sarbecovirus of betacoronaviruses.

[Vetch poisoning in a Bavarian cattle herd - A case report from a pathological perspective]. / Wickenvergiftung in einem bayerischen Rinderbestand.

In a herd of 40 cattle of the Holstein-Friesian breed, 6 animals were euthanized for severe pruritus, fever and skin lesions. Pathomorphological examination of 3 animals revealed multisystemic granulomatous inflammation involving multinuclear giant cells. The content of vetch in the feed used and the characteristic histology led to the diagnosis of vetch poisoning.

Co-infections: Simultaneous detections of West Nile virus and Usutu virus in birds from Germany.

The emergence of West Nile virus (WNV) and Usutu virus (USUV) in Europe resulted in significant outbreaks leading to avifauna mortality and human infections. Both viruses have overlapping geographical, host and vector ranges, and are often co-circulating in Europe. In Germany, a nationwide bird surveillance network was established to monitor these zoonotic arthropod-borne viruses in migratory and resident birds. In this framework, co-infections with WNV and USUV were detected in six dead birds collected in 2018 and 2019. Genomic sequencing and phylogenetic analyses classified the detected WNV strains as lineage 2 and the USUV strains as lineages Africa 2 (n = 2), Africa 3 (n = 3) and Europe 2 (n = 1). Preliminary attempts to co-propagate both viruses in vitro failed. However, we successfully cultivated WNV from two animals. Further evidence for WNV-USUV co-infection was obtained by sampling live birds in four zoological gardens with confirmed WNV cases. Three snowy owls had high neutralizing antibody titres against both WNV and USUV, of which two were also positive for USUV-RNA. In conclusion, further reports of co-infections in animals as well as in humans are expected in the future, particularly in areas where both viruses are present in the vector population.

Prolonged SARS-CoV-2 RNA Shedding from Therapy Cat after Cluster Outbreak in Retirement Home.

We report a therapy cat in a nursing home in Germany infected with severe acute respiratory syndrome coronavirus 2 during a cluster outbreak in the home residents. Although we confirmed prolonged presence of virus RNA in the asymptomatic cat, genome sequencing showed no further role of the cat in human infections on site.

Exposure of wild boar to Influenza A viruses in Bavaria: Analysis of seroprevalences and antibody subtype specificity before and after the panzootic of highly pathogenic avian influenza viruses A (H5N8).

Swine influenza A viruses (S-IAV) circulate in wild boar populations worldwide. Subtypes primarily reflect those actually present within the respective pig industry. Accordingly, infections with swine H1N1, H1N2 and H3N2 have been reported for several regions of Germany. As pigs are susceptible not only to S-IAV but also to avian and human influenza A viruses, it is necessary to consider the possibility that new reassortant viruses with pandemic potential may arise in these new hosts. Therefore, in this study the impact of recent IAV epidemics on antibody prevalences in Bavarian wild boar was assessed. Important events considered were the H1N1pdm09 pandemic, which affected humans and swine, and the highly pathogenic avian influenza (HPAI) H5N8 panzootic in 2016 and 2017, affecting wild and domestic birds. IAV seroprevalences were determined analysing 1,396 samples from before and after the H5N8 panzootic, from various regions in Bavaria, a large administrative region in the South of Germany. Taken together, seroprevalences varied markedly from 1.44% to 12.59%, relative to region and time. However, no discrete correlation was found to population density either in wild boar or in pigs. Antibodies against H1N1 were the most prevalent. In addition, antibodies were detected reacting against H1N2 and against H1pdmNx reassortant viruses, already known to circulate in domestic pigs in Bavaria and notably also against the avian influenza A virus H5N8; the latter in samples taken in 2017. These results confirm the exposure of wild boar to IAV of diverse origin and the increasing variability of S-IAV present in the field. The necessity for continuous IAV surveillance not only of domestic swine but also of wildlife is emphasized.

Comparison of nine different commercially available molecular assays for detection of SARS-CoV-2 RNA.

To face the COVID-19 pandemic, the need for fast and reliable diagnostic assays for the detection of SARS-CoV-2 is immense. We describe our laboratory experiences evaluating nine commercially available real-time RT-PCR assays. We found that assays differed considerably in performance and validation before routine use is mandatory.

Full genome sequence of bovine alphaherpesvirus 2 (BoHV-2).

We present the complete genome sequence of bovine alphaherpesvirus 2 (BoHV-2), a member of the family Herpesviridae, subfamily Alphaherpesvirinae, genus Simplexvirus. BoHV-2 is the causative agent of bovine ulcerative mammillitis (bovine herpes mammillitis) and pseudo-lumpy skin disease. The genomic architecture of BoHV-2 is typical of most simplexvirus genomes and congruent with that of human alphaherpesvirus 1 (HHV-1). The genome comprises a total of 131,245 base pairs and has an overall G+C content of 64.9 mol%. A total of 75 open reading frames are predicted. The gene repertoire of BoHV-2 is analogous to that of HHV-1, although the coding region of US12 is missing. A phylogenetic analysis supported BoHV-2 as a member of the genus Simplexvirus.

Bovine Alphaherpesvirus 2 infections in Bavaria: an analysis of the current situation - several years after eradicating Bovine Alphaherpesvirus 1.

BACKGROUND: Bavaria, a large federal state in Germany, has been declared free from infections with Bovine Alphaherpesvirus 1 (BoHV-1) in 2011. To maintain this status the cattle population is monitored for antibodies against BoHV-1 regularly. Several years ago, infrequent but recurrent problems in this sero-surveillance were statistically put into correlation with the presence of antibodies against Bovine Alphaherpesvirus 2 (BoHV-2). In Europe, BoHV-2 is primarily known as the agent causing bovine herpes mammillitis. However, very little information about BoHV-2 infections in Bavaria is available so far. Therefore, the aims of this study were to determine BoHV-2 seroprevalences and to detect virus genomes in potential clinical samples. RESULTS: 6801 blood sera of healthy cattle from all over Bavaria were tested for antibodies against BoHV-2, revealing an overall seroprevalence of 5.51%. Interestingly, seroprevalences markedly varied between the North and the South of Bavaria, namely from 0.42 to 11.17%. Concurrently, the previously reported relation between the epidemiologically inexplicable sero-reactivities in BoHV-1 ELISAs and the presence of BoHV-2 infections were statistically corroborated in this study. To detect BoHV-2 genomes a fast and sensitive real time PCR was established. Using a multiple PCR strategy, tissue samples from skin lesions at relevant localizations, corresponding lymph nodes, and trigeminal ganglia from 111 animals, as well as nasal swabs from 918 bovines with respiratory symptoms were tested. However, BoHV-2 genomes were not detected in any of these samples. CONCLUSIONS: BoHV-2 antibodies were found in samples from bovines all over Bavaria, albeit with an explicit South-North-divide. BoHV-2 genomes, however, could not be detected in any of the analyzed samples, indicating that acute clinical cases as well as obvious virus reactivation are relatively rare. Consequently, the future spread of BoHV-2 infections throughout Bavaria, particularly, after eradicating BoHV-1, has to be further monitored.

Molecular Epidemiology and Evolution of European Bat Lyssavirus 2.

Bat rabies cases in Europe are mainly attributed to two lyssaviruses, namely European Bat Lyssavirus 1 (EBLV-1) and European Bat Lyssavirus 2 (EBLV-2). Prior to the death of a bat worker in Finland in 1985, very few bat rabies cases were reported. Enhanced surveillance in the two subsequent years (1986-1987) identified 263 cases (more than a fifth of all reported cases to date). Between 1977 and 2016, 1183 cases of bat rabies were reported, with the vast majority (>97%) being attributed to EBLV-1. In contrast, there have been only 39 suspected cases of EBLV-2, of which 34 have been confirmed by virus typing and presently restricted to just two bat species; Myotis daubentonii and Myotis dasycneme. The limited number of EBLV-2 cases in Europe prompted the establishment of a network of European reference laboratories to collate all available viruses and data. Despite the relatively low number of EBLV-2 cases, a large amount of anomalous data has been published in the scientific literature, which we have here reviewed and clarified. In this review, 29 EBLV-2 full genome sequences have been analysed to further our understanding of the diversity and molecular evolution of EBLV-2 in Europe. Analysis of the 29 complete EBLV-2 genome sequences clearly corroborated geographical relationships with all EBLV-2 sequences clustering at the country level irrespective of the gene studied. Further geographical clustering was also observed at a local level. There are high levels of homogeneity within the EBLV-2 species with nucleotide identities ranging from 95.5-100% and amino acid identities between 98.7% and 100%, despite the widespread distribution of the isolates both geographically and chronologically. The mean substitution rate for EBLV-2 across the five concatenated genes was 1.65 × 10-5, and evolutionary clock analysis confirms the slow evolution of EBLV-2 both between and within countries in Europe. This is further supported by the first detailed EBLV-2 intra-roost genomic analysis whereby a relatively high sequence homogeneity was found across the genomes of three EBLV-2 isolates obtained several years apart (2007, 2008, and 2014) from M. daubentonii at the same site (Stokesay Castle, Shropshire, UK).

The Recently Discovered Bokeloh Bat Lyssavirus: Insights Into Its Genetic Heterogeneity and Spatial Distribution in Europe and the Population Genetics of Its Primary Host.

In 2010, a novel lyssavirus named Bokeloh bat lyssavirus (BBLV) was isolated from a Natterer's bat (Myotis nattereri) in Germany. Two further viruses were isolated in the same country and in France in recent years, all from the same bat species and all found in moribund or dead bats. Here we report the description and the full-length genome sequence of five additional BBLV isolates from Germany (n=4) and France (n=1). Interestingly, all of them were isolated from the Natterer's bat, except one from Germany, which was found in a common Pipistrelle bat (Pipistrellus pipistrellus), a widespread and abundant bat species in Europe. The latter represents the first case of transmission of BBLV to another bat species. Phylogenetic analysis clearly demonstrated the presence of two different lineages among this lyssavirus species: lineages A and B. The spatial distribution of these two lineages remains puzzling, as both of them comprised isolates from France and Germany; although clustering of isolates was observed on a regional scale, especially in Germany. Phylogenetic analysis based on the mitochondrial cytochrome b (CYTB) gene from positive Natterer's bat did not suggest a circulation of the respective BBLV sublineages in specific Natterer's bat subspecies, as all of them were shown to belong to the M. nattereri sensu stricto clade/subspecies and were closely related (German and French positive bats). At the bat host level, we demonstrated that the distribution of BBLV at the late stage of the disease seems large and massive, as viral RNA was detected in many different organs.

A two year BTV-8 vaccination follow up: molecular diagnostics and assessment of humoral and cellular immune reactions.

The compulsory vaccination campaign against Bluetongue virus serotype eight (BTV-8) in Germany was exercised in the state of Bavaria using three commercial monovalent inactivated vaccines given provisional marketing authorisation for emergency use. In eleven Bavarian farms representing a cross sectional area of the state the immune reactions of sheep and cattle were followed over a two year period (2008-2009) using cELISA, a serum neutralisation test (SNT) and interferon gamma (IFN-γ) ELISPOT. For molecular diagnostics of BTV genome presence two recommended real time quantitative RT-PCR protocols were applied. The recommended vaccination scheme led to low or even undetectable antibody titers (ELISA) in serum samples of both cattle and sheep. A fourfold increase of the vaccine dose in cattle, however, induced higher ELISA titers and virus neutralising antibodies. Accordingly, repeated vaccination in sheep caused an increase in ELISA-antibody titers. BTV-8 neutralising antibodies occurred in most animals only after multiple vaccinations in the second year of the campaign. The secretion of interferon gamma (IFN-γ) in ELISPOT after in vitro re-stimulation of PBMC of BTV-8 vaccinated animals with BTV was evaluated in the field for the first time. Sera of BTV-8 infected or vaccinated animals neutralising BTV-8 could also neutralise an Italian BTV serotype 1 cell culture adapted strain and PBMC of such animals secreted IFN-γ when stimulated with BTV-1.

[Surveillance of wild birds for avian influenza A virus (AIV) in Bavaria in the years 2007 and 2008]. / Wildvogelmonitoring zum Vorkommen von aviärem Influenza A Virus (AIV) in Bayern in den Jahren 2007 und 2008.

A monitoring programme has been initiated in Bavaria to continuously control wild birds for the presence of avian Influenza A virus (AIV) and to monitor the possible occurrence and accumulation of notifiable AIV subtypes as an early-warning system. In addition information about the regional, seasonal and species-specific distribution of AIV could be obtained. Between July 2007 and December 2008 samples from 5864 wild birds of twelve different zoological orders had been collected (cloacal- and tracheal swab samples, droppings, and organs) and analysed. AIV genomes were detected in 3.7% of the 5864 wild birds by RT real time PCR. The subtype component H5 was identified in 52 samples (0.9%) and the N1 subtype component in 13 samples (0.2%), but never in combination with each other. The hemagglutinine subtype component H7 could not be detected. Most of the positive AIV genome results originated from samples in the district Swabia, which is situated in the central area of the south-west bird migration route across southern Germany and harbours favourable resting areas for migrating birds. Mallards (Anas platyrhynchos) were the most frequently sampled bird species and had the highest AIV infection rate of 6.4%, followed by Tufted ducks (Aythya fuligula) (AIV prevalence of 5.4%), Mute Swans (Cygnus olor) (1.6%), Coots (Fulica atra) (0.3%) and Greylag Goose (Anser anser) (0.1%). The detection rate of AIV in Bavarian wild birds showed a seasonal peak in autumn/winter. Ten virus isolates could be obtained after sample inoculation in embryonated hen's eggs.

Highly pathogenic avian influenza virus (H5N1) in frozen duck carcasses, Germany, 2007.

We conducted phylogenetic and epidemiologic analyses to determine sources of outbreaks of highly pathogenic avian influenza virus (HPAIV), subtype H5N1, in poultry holdings in 2007 in Germany, and a suspected incursion of HPAIV into the food chain through contaminated deep-frozen duck carcasses. In summer 2007, HPAIV (H5N1) outbreaks in 3 poultry holdings in Germany were temporally, spatially, and phylogenetically linked to outbreaks in wild aquatic birds. Detection of HPAIV (H5N1) in frozen duck carcass samples of retained slaughter batches of 1 farm indicated that silent infection had occurred for some time before the incidental detection. Phylogenetic analysis established a direct epidemiologic link between HPAIV isolated from duck meat and strains isolated from 3 further outbreaks in December 2007 in backyard chickens that had access to uncooked offal from commercial deep-frozen duck carcasses. Measures that will prevent such undetected introduction of HPAIV (H5N1) into the food chain are urgently required.