Suttonella ornithocola detected within lesions of tit birds (Paridae) from epidemic death episodes in Germany, 2018-2020.

Several episodes of increased mortality in wild birds of the families Paridae and Aegithalidae have been documented in recent decades. The majority of affected animals exhibited necrotizing pneumonia with intralesional bacteria. Suttonella (S.) ornithocola, a gram-negative bacterium in the Cardiobacteriaceae family, has been regularly cultured bacteriologically from affected birds and has long been suspected as a potentially fatal cause of respiratory disease in birds. However, a direct causal relationship between this specific bacterium and the observed lesions within birds has not yet been established. Therefore, postmortem tissue from six tits was used in the present study, including three blue tits (Cyanistes caeruleus) and three great tits (Parus major). Five of the six tits tested positive for S. ornithocola in bacteriological examination and originated from two incidents of increased mortality in Paridae in Germany. Animals found dead in the administrative district of Arnsberg (North Rhine Westphalia) in 2018 and 2020 were investigated for genomic fragments of S. ornithocola by chromogenic in situ hybridization using a newly developed DNA probe based on publicly assessable DNA sequences of the 16S rRNA gene of S. ornithocola. Positive hybridization signals were detected in five out of five animals and were predominantly detected within necrotizing lesions in lung and occasionally in lesions affecting liver and trachea. Interestingly, the lung of one animal without obvious necrotizing pulmonary lesions revealed positive hybridization results in the lumen of one pulmonary blood vessel. Two negative controls, including one bacteriologically S. ornithocola-negative great tit and a cattle egret (Bubulcus ibis) suffering from salmonellosis, did not yield positive signals, indicating high sensitivity and specificity of the probe used. This is the first time that S. ornithocola has been clearly identified within necrotizing lesions in deceased tits. Although Koch's postulates have yet to be fulfilled, positive hybridization signals in association with detectable lesions are considered as further and strong evidence of the significant contribution of S. ornithocola to the several episodes of tit mortality recorded in Germany.

In action-an early warning system for the detection of unexpected or novel pathogens.

Proactive approaches in preventing future epidemics include pathogen discovery prior to their emergence in human and/or animal populations. Playing an important role in pathogen discovery, high-throughput sequencing (HTS) enables the characterization of microbial and viral genetic diversity within a given sample. In particular, metagenomic HTS allows the unbiased taxonomic profiling of sequences; hence, it can identify novel and highly divergent pathogens such as viruses. Newly discovered viral sequences must be further investigated using genomic characterization, molecular and serological screening, and/or in vitro and in vivo characterization. Several outbreak and surveillance studies apply unbiased generic HTS to characterize the whole genome sequences of suspected pathogens. In contrast, this study aimed to screen for novel and unexpected pathogens in previously generated HTS datasets and use this information as a starting point for the establishment of an early warning system (EWS). As a proof of concept, the EWS was applied to HTS datasets and archived samples from the 2018-9 West Nile virus (WNV) epidemic in Germany. A metagenomics read classifier detected sequences related to genome sequences of various members of Riboviria. We focused the further EWS investigation on viruses belonging to the families Peribunyaviridae and Reoviridae, under suspicion of causing co-infections in WNV-infected birds. Phylogenetic analyses revealed that the reovirus genome sequences clustered with sequences assigned to the species Umatilla virus (UMAV), whereas a new peribunyavirid, tentatively named 'Hedwig virus' (HEDV), belonged to a putative novel genus of the family Peribunyaviridae. In follow-up studies, newly developed molecular diagnostic assays detected fourteen UMAV-positive wild birds from different German cities and eight HEDV-positive captive birds from two zoological gardens. UMAV was successfully cultivated in mosquito C6/36 cells inoculated with a blackbird liver. In conclusion, this study demonstrates the power of the applied EWS for the discovery and characterization of unexpected viruses in repurposed sequence datasets, followed by virus screening and cultivation using archived sample material. The EWS enhances the strategies for pathogen recognition before causing sporadic cases and massive outbreaks and proves to be a reliable tool for modern outbreak preparedness.

Salmonella enterica subspecies enterica serovar Choleraesuis in a German wild boar population: occurrence and characterisation.

BACKGROUND: The swine-adapted serovar Choleraesuis of Salmonella enterica subspecies enterica is found rarely in domestic pigs in Germany. However, a considerable and increasing number of S. Choleraesuis organisms have been isolated from wild boars in Germany in recent years. To investigate a possible epidemiological context, S. Choleraesuis strains from a regional German wild boar population and other hosts were characterised by genotyping methods. RESULTS: Macrorestriction analysis, biochemical differentiation and antimicrobial susceptibility typing enabled the identification of several clusters of S. Choleraesuis. Some clusters occurred almost permanently in a certain district, whereas other groups circulated among different wild boar herds in larger regions. Non-porcine hosts were infected with the same cluster as the wild boars. CONCLUSIONS: The emergence of S. Choleraesuis in wild boars might be caused by a higher prevalence in the wild boar population, but also the higher awareness to infections with African swine fever may have resulted in a higher number of examined animals. Separation of wild boar populations and, as a result, also the diverse S. Choleraesuis organisms might be due to natural barriers and artificial barriers like arterial roads. The findings of S. Choleraesuis in domestic pigs emphasize the importance of strict biosecurity measures to prevent transmission from wild boars of this but also other pathogens. To avoid risks for humans by zoonotic pathogens regular inspections of meat from wildlife need to be conducted.

Effects of induced endometritis on uterine blood flow in cows as evaluated by transrectal Doppler sonography.

This study was conducted to evaluate the effects of induced endometritis on uterine blood flow in cows. Transrectal Doppler sonography was performed on uterine arteries of six cyclic cows before and for 4 days after inducing acute endometritis by intrauterine infusion of 720 mg of policresulen, and for 4 days of the following estrous cycle. Time-averaged maximum velocity (TAMV) increased (p < 0.001) and pulsatility index (PI) decreased (p < 0.0001) within 1 h of policresulen administration, and did not change (p > 0.05) in the next 4 days of the same cycle. TAMV and PI values in the subsequent cycle did not differ (p > 0.05) from the values measured before infusion and showed no changes (p > 0.05) within the cycle. Blood flow parameters were not related (p > 0.05) to plasma concentrations of progesterone and estrogen. All cows showed an acute endometritis determined by histopathological findings of biopsy samples taken 1 day after infusion and fibrotic endometrial alterations detected in the subsequent cycle. No relationships were observed between fibrotic changes of the endometrium and uterine blood flow during either cycle. In conclusion, acute inflammation is accompanied by a rise in uterine blood flow, but fibrotic alterations do not seem to be related to Doppler sonographic findings.