The sudden emergence of SARS-CoV-2 demonstrates the need for new vaccines that rapidly protect in the case of an emergency. In this study, we developed a recombinant MVA vaccine co-expressing SARS-CoV-2 prefusion-stabilized spike protein (ST) and SARS-CoV-2 nucleoprotein (N, MVA-SARS-2-ST/N) as an approach to further improve vaccine-induced immunogenicity and efficacy. Single MVA-SARS-2-ST/N vaccination in K18-hACE2 mice induced robust protection against lethal respiratory SARS-CoV-2 challenge infection 28 days later. The protective outcome of MVA-SARS-2-ST/N vaccination correlated with the activation of SARS-CoV-2-neutralizing antibodies (nABs) and substantial amounts of SARS-CoV-2-specific T cells especially in the lung of MVA-SARS-2-ST/N-vaccinated mice. Emergency vaccination with MVA-SARS-2-ST/N just 2 days before lethal SARS-CoV-2 challenge infection resulted in a delayed onset of clinical disease outcome in these mice and increased titers of nAB or SARS-CoV-2-specific T cells in the spleen and lung. These data highlight the potential of a multivalent COVID-19 vaccine co-expressing S- and N-protein, which further contributes to the development of rapidly protective vaccination strategies against emerging pathogens.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), is known to affect multiple organ systems, including the respiratory tract and nervous and ocular systems. This retrospective study aimed to characterize the spatiotemporal distribution of viral antigen and associated pathological changes in the nose, lungs, brain, and eyes of K18-hACE2 mice intranasally infected with SARS-CoV-2. Using histology and immunohistochemistry, tissues were examined at 3, 6, and 7/8 days post-infection (dpi). In addition, lung and brain tissues were analyzed by means of RT-qPCR to determine viral RNA titers. Viral antigen was most pronounced in the nose, brain, and lung at 3, 6, and 7/8 dpi, respectively, whereas viral antigen was detected at 6 and 7/8 dpi in the retina. Quantitative PCR confirmed increasing viral RNA levels in both lung and brain, peaking at 7/8 dpi. Nasal and lung inflammation mirrored viral antigen distribution and localization. In the brain, the predominantly basal viral spread correlated with lymphohistiocytic meningoencephalitis, neuronal vacuolation, and altered neurofilament immunoreactivity. Retinal ganglion cells showed viral antigen expression without associated lesions. Microglial activation was evident in both the optic chiasm and the brain. These findings highlight the K18-hACE2 model’s utility for studying extrapulmonary SARS-CoV-2 pathogenesis. Understanding the temporal and spatial dynamics of viral spread enhances insights into SARS-CoV-2 neurotropism and its clinical manifestations.

The Eurasian lynx (Lynx lynx), a widespread wild felid on the Eurasian continent, is currently classified as “critically endangered” in Germany. Understanding the impact of infectious agents is of particular importance for the continued conservation of these animals, especially regarding pathogens with broad host ranges and risk of interspecies transmission. Feline leukemia virus (FeLV) is known to infect wild and domestic felids worldwide, including several species of lynx, but it has not been reported thus far in the Eurasian lynx. In September 2020, a 16-month-old female Eurasian lynx from the Bavarian Forest, Germany, showed a sudden onset of gastrointestinal signs such as anorexia, diarrhea, and vomiting, and died within one week. Macroscopic and histologic examination revealed hemorrhagic-necrotizing enteritis and typhlocolitis, with the degeneration of crypts and crypt abscesses, as well as depleted Peyer’s patches. In addition, the animal showed lymphoid depletion (lymph nodes, thymus, and spleen) and hypocellularity of the bone marrow. FeLV infection was confirmed by immunohistochemistry and next generation sequencing. A secondary bacterial infection with hemolytic Escherichia coli and Clostridium perfringens type A was present in the intestine. This is the first report of FeLV-associated enteritis, lymphoid depletion and bone marrow suppression with associated secondary bacterial infection in a Eurasian lynx.

Native to South America, the coypu (Myocastor coypus) is an invasive alien species (IAS) of Union concern. It was introduced to Germany a hundred years ago and is considered established in all German federal states. Between January and February 2021, ground temperatures below −10° Celsius were recorded in Lower Saxony, Germany, for approximately two consecutive weeks. Five male and five female coypus, harvested between 23 February and 31 March 2021, received a post-mortem examination. Nutritional status was poor in six cases, moderate in three and good in one case. Pregnancy was observed in two females. In all the animals, lesions were predominantly found on the distal limbs (n = 7) and/or tail (n = 10), involving the skin and soft tissue with occasional exposure or loss of bones. The histological findings consisted of chronic, ulcerative to necrotizing dermatitis and occasional ulcerative-suppurative dermatitis, necrotizing myositis, thrombosis, granulation tissue, fibrosis and intralesional dystrophic mineralization. Intralesional bacteria were present in six and fungal spores in one animal. Determination of the exact cause was not possible; however, considering the local weather conditions and the distribution of lesions, frostbite injuries have to be considered as the most likely cause. The intralesional bacteria and fungal spores most likely represent secondary contaminants. Interestingly, lesions of this kind have not been reported in coypus in Germany so far. Therefore, frostbite should be considered as a potential cause of disease in coypus, warranting further investigation.

Neuronal ceroid lipofuscinoses (NCL) are among the most prevalent neurodegenerative disorders of early life in humans. Disease‐causing variants have been described for 13 different NCL genes. In this study, a refined pathological characterization of a female rabbit with progressive neurological signs reminiscent of NCL was performed. Cytoplasmic pigment present in neurons was weakly positive with Sudan black B and autofluorescent. Immunohistology revealed astrogliosis, microgliosis and axonal degeneration. During the subsequent genetic investigation, the genome of the affected rabbit was sequenced and examined for private variants in NCL candidate genes. The analysis revealed a homozygous ~10.7 kb genomic duplication on chromosome 15 comprising parts of the MFSD8 gene, NC_013683.1:g.103,727,963_103,738,667dup. The duplication harbors two internal protein coding exons and is predicted to introduce a premature stop codon into the transcript, truncating ~50% of the wild‐type MFSD8 open reading frame encoding the major facilitator superfamily domain containing protein 8, XP_002717309.2:p.(Glu235Leufs*23). Biallelic loss‐of‐function variants in MFSD8 have been described to cause NCL7 in human patients, dogs and a single cat. The available clinical and pathological data, together with current knowledge about MFSD8 variants and their functional impact in other species, point to the MFSD8 duplication as a likely causative defect for the observed phenotype in the affected rabbit.

Arthropod-borne diseases represent one of the greatest infection-related threats as a result of climate change and globalization. Repeatedly, arbovirus-infected mosquitoes show behavioral changes whose underlying mechanisms are still largely unknown, but might help to develop control strategies. However, in contrast to well-characterized insects such as fruit flies, little is known about neuroanatomy and neurotransmission in mosquitoes. To overcome this limitation, the study focuses on the immunohistochemical characterization of the nervous system of Culex pipiens biotype molestus in comparison to Drosophila melanogaster using 13 antibodies labeling nervous tissue, neurotransmitters or neurotransmitter-related enzymes. Antibodies directed against γ-aminobutyric acid, serotonin, tyrosine-hydroxylase and glutamine synthetase were suitable for investigations in Culex pipiens and Drosophila melanogaster, albeit species-specific spatial differences were observed. Likewise, similar staining results were achieved for neuronal glycoproteins, axons, dendrites and synaptic zones in both species. Interestingly, anti-phosphosynapsin and anti-gephyrin appear to represent novel markers for synapses and glial cells, respectively. In contrast, antibodies directed against acetylcholine, choline acetyltransferase, elav and repo failed to produce a signal in Culex pipiens comparable to that in Drosophila melanogaster. In summary, present results enable a detailed investigation of the nervous system of mosquitoes, facilitating further studies of behavioral mechanisms associated with arboviruses in the course of vector research.