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The genus Sporothrix contains both species pathogenic to humans and animals as well as environmental fungi. S. humicola, a member of the latter S. pallida clade, has previously been reported only from soil. We have isolated this dimorphic fungus from multiple cutaneous lesions in two endangered marsupials native to Tasmania. Clinical appearance resembled cutaneous sporotrichosis, highlighting the principle pathogenic potential. Identification was based on partial ITS, β-tubulin and calmodulin gene sequencing. Keywords: Sporothrix humicola, Eastern quoll, Dasyurus viverrinus, Dermatomycosis, MALDI-TOF MS

A number of different Chlamydia spp. have been detected in the class Amphibia with C. pneumoniae being the predominant species involved. Chlamydiae have been linked to mass mortality events, thereby representing significant pathogens that deserve attention with respect to worldwide amphibian decline. We here present six cases of chlamydiosis and asymptomatic chlamydial infections in different frog species from three ex situ amphibian conservation facilities. Clinical signs predominantly characterised by regurgitation, chronic wasting, lethargy and suspended breeding were associated with C. pneumoniae infection. Despite various treatment regimens, it was not possible to clear infections. However, intra vitam diagnostics succeeded from skin, faeces and urine for the first time.

Live traps are established in pest control, fishing and hunting, but they are discussed to compromise animal welfare due to inadequate construction and control of the trap. To assure animal welfare, the Agreement on International Humane Trapping Standards (AIHTS) demands for legal regulations regarding trapping devices. According to AIHTS, the certification of each trap is based on evaluation in 20 individuals of a specific animal species (target species). To this end, more than 80% of the animals must not show specific alterations of physiology and behaviour or potential injuries. The aim of this study was to evaluate the ‘Krefelder Fuchsfalle’, equipped with an electronic trap system, according to AIHTS standards. The raccoon (Procyon lotor) was chosen as target species, as it is an invasive species in urban regions of Central Europe, a potential vector for zoonotic pathogens, and a skilful animal, which potentially is most difficult to meet the AIHTS requirements. The behaviour of the trapped animal was evaluated, before shooting it and performing radiographical, pathological and histological examination. In total, 20 raccoons were trapped within 10 months in a hunting district in Hesse, Germany. Mean period of stay within the trap were 6.39 h. Despite of 14 animals, which demonstrated low-grade excoriations (superficial skin layer) at the paws and rhinarium, no other injuries or alterations of physiology and behaviour were detected. Therefore, this study demonstrates that the ‘Krefelder Fuchsfalle’ fulfilled the AIHTS standards and accomplished trapping according to animal welfare.

SUMMARYGermany has been an officially bovine tuberculosis (bTB)-free (OTF) country since 1996. Gradually rising numbers of bTB herd incidents due toMycobacterium bovisandM. capraein North-Western and Southern Germany during the last few years prompted the competent authorities to conduct a nationwide bTB survey in 2013/2014. This led to the detection of a dairy herd in which as many as 55 cattle reacted positively to consecutiveintra vitamtesting. Test-positive animals lacked visible lesions indicative of bTB at necropsy. Extensive mycobacterial culturing as well as molecular testing of samples from 11 tissues for members of theM. tuberculosiscomplex (MTC) yielded negative results throughout. However, caseous lymphadenitis of Ln. mandibularis accessorius was observed during meat inspection of a fattening pig from the same farm at regular slaughter at that time. Respective tissue samples tested MTC positive by polymerase chain reaction, andM. tuberculosisT1 family were identified by spoligotyping. Four human reactors within the farmer's family were also found to be immunoreactive. As exposure of livestock toM. tuberculosisis not generally considered, its impact may result in regulatory and practical difficulties when using protocols designed to detect classical bTB, particularly in OTF countries.

Background Next to various known infectious and non-infectious causes, the aetiology of non-suppurative encephalitis in red foxes (Vulpes vulpes) often remains unclear. Known causes in foxes imply rabies, canine distemper, toxoplasmosis, Aujeszky’s disease, as well as parvovirus, adenovirus, circovirus and flavivirus infections. In this study, particular attention was paid on bornaviruses, since red foxes are predators of bicoloured white-toothed shrews, a reservoir of Borna disease virus 1 (BoDV-1). In addition, foxes are known to be highly susceptible for viruses of the order Mononegavirales. Methods Analyses for the presence of anti-BoDV-1 antibodies, BoDV-1-RNA and antigen were performed on 225 blood and 59 brain samples, from a total of 232 red foxes. Foxes originated from BoDV-1 endemic and non-endemic German areas. Additional investigations for the presence of rabies, canine distemper, toxoplasmosis, Aujeszky’s disease, parvovirus, adenovirus and flavivirus infections were carried out on 16 red foxes with non-suppurative (meningo-) encephalitis. A metagenomic analysis was used on three representative brain samples displaying encephalitis. Results Among 225 foxes, 37 displayed anti-BoDV-1 antibodies with titres ranging between 1:40 and 1:2560, regardless of geographic origin. In 6 out of 16 foxes with encephalitis, canine distemper virus was detected. No evidence of any of the other investigated agents was found in the 16 fox brains with encephalitis. Metagenomics revealed no infectious agents, except for one already known canine distemper case. Conclusion Red foxes can exhibit BoDV-1 specific antibodies without association with geographic origin or encephalitis due to bornavirus infection. The encephalitis pattern was highly conspicuous for a viral infection, but remained unclear in 10 out of 16 foxes. Thus, presently unknown infectious and non-infectious causes need to be considered and further investigated, especially since foxes also tend to occur in human proximity.

In a breeding and fattening pig farm an increasing number of cases of abortion and generalized mycobacteriosis at slaughter occurred. Pathological findings compatible with mycobacteriosis, acid-fast organisms in tissues, and isolation of mycobacteria from tissue samples including fetuses, lungs and reproductive organs from sows, genital swabs, mesenteric lymph nodes, and from a sperm sample revealed the cause of the disease. Bacterial cultures were identified as Mycobacterium avium subsp. hominissuis using IS901-/ IS1245-specific PCR. Genotyping of selected isolates from animals as well as from their environment by MIRU-VNTR analysis showed that the herd was infected with one single outbreak strain. The same genotype was also isolated from pigs of two other farms which showed comparable symptoms and were in direct contact with the index farm as well as from their environment. Immunological host responses detected by tuberculin skin test and ELISA gave positive results at herd level only. Despite the detection of other potential pathogens mycobacteria were regarded as the causative agent of the reproductive disorders. To our knowledge this is the first report of an epidemic mycobacterial infection in a pig holding associated with reproductive disorders, which could be attributed to one single virulent strain, and the first report of detection of M. avium subsp. hominissuis in pig sperm.

Introduction: Streptobacillus moniliformis causes rat‐bite fever, an underdiagnosed zoonosis occurring worldwide. A variety of animals including livestock and exotic mammals are known to be susceptible hosts for this species, but little information is available regarding infection in companion animals. Case presentation: Following the necropsy of a domestic cat, bacteria displaying substantial characteristics of Streptobacillus sp. were cultured from pneumonic lung tissue. Streptobacillus‐like morphological features observed included strictly microaerophilic pleomorphic Gram‐negative rods with bulbar swellings that grew exclusively in the presence of serum. Significant shared biochemical properties included negative reactions for cytochrome oxidase, catalase, urease, nitrate reduction and indole production, as well as broad antimicrobial susceptibility. These characteristics are all indicative of Streptobacillus moniliformis. However, 16S rRNA gene sequencing revealed only 98 % sequence homology to type strain DSM 12112. A mass spectrometry analysis confirmed the affiliation of the domestic cat isolate described in this study with bacteria of the genus Streptobacillus, but matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry indicated that it differed from nine reference strains of Streptobacillus moniliformis isolated from various sources and host species. Conclusion: This is the first evidence for clinical disease caused by a streptobacillary infection in a domestic cat.

Coronary atherectomy specimens from 50 patients with coronary heart disease were examined for the presence of Chlamydia pneumoniae by two different methods of polymerase chain reaction (PCR) and by in situ hybridization. C. pneumoniae DNA was detected by PCR in atherosclerotic plaques of four patients (8%). Two patients' coronary atheromas were positive, both by a single-step 16S rRNA-based PCR and by an omp1-based nested PCR. The other two patients' specimens were positive only by the nested PCR. In contrast, C. pneumoniae was not detected by in situ hybridization in any of the cardiovascular tissues tested. Of three patients with evidence of C. pneumoniae in coronary atheromas, two had an antibody titer of 1:32 and the third had no specific antibodies detectable. Results of this study demonstrate a low prevalence of C. pneumoniae DNA in coronary atheromas. These findings do not support the hypothesis that the organism plays a major role in atherogenesis.

Background Tularemia is a zoonotic disease caused by Francisella tularensis that has been found in many different vertebrates. In Germany most human infections are caused by contact with infected European brown hares (Lepus europaeus). The aim of this study was to elucidate the epidemiology of tularemia in hares using phenotypic and genotypic characteristics of F. tularensis. Results Cultivation of F. tularensis subsp. holarctica bacteria from organ material was successful in 31 of 52 hares that had a positive PCR result targeting the Ft-M19 locus. 17 isolates were sensitive to erythromycin and 14 were resistant. Analysis of VNTR loci (Ft-M3, Ft-M6 and Ft-M24), INDELs (Ftind33, Ftind38, Ftind49, RD23) and SNPs (B.17, B.18, B.19, and B.20) was shown to be useful to investigate the genetic relatedness of Francisella strains in this set of strains. The 14 erythromycin resistant isolates were assigned to clade B.I, and 16 erythromycin sensitive isolates to clade B.IV and one isolate was found to belong to clade B.II. MALDI-TOF mass spectrometry (MS) was useful to discriminate strains to the subspecies level. Conclusions F. tularensis seems to be a re-emerging pathogen in Germany. The pathogen can easily be identified using PCR assays. Isolates can also be identified within one hour using MALDI-TOF MS in laboratories where specific PCR assays are not established. Further analysis of strains requires genotyping tools. The results from this study indicate a geographical segregation of the phylogenetic clade B.I and B.IV, where B.I strains localize primarily within eastern Germany and B.IV strains within western Germany. This phylogeographical pattern coincides with the distribution of biovar I (erythromycin sensitive) and biovar II (erythromycin resistance) strains. When time and costs are limiting parameters small numbers of isolates can be analysed using PCR assays combined with DNA sequencing with a focus on genetic loci that are most likely discriminatory among strains found in a specific area. In perspective, whole genome data will have to be investigated especially when terrorist attack strains need to be tracked to their genetic and geographical sources.