Wencke Reineking

All Publications

• Strain-resolved metagenomic analysis and qPCR validation suggest Clostridium cuniculi is the etiologic agent for infectious diarrhea in severely immunodeficient mice. Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc Barouch-Bentov, R., Merrill, B. D., Reineking, W., Moorhead, R., Herberg de Alonso, F., Fazel, M., Uzal, F. A., Felt, S. A., Sonnenburg, J. L., Casey, K. M., Nagamine, C. M. 2025: 10406387251378688

  Abstract

  In late 2020, the mouse barrier facility at Stanford University experienced an outbreak of diarrhea in adult mice and sudden deaths in mid-lactation females. Affected strains were immunodeficient, carrying either the Prkdcscid or Ragnull mutations and the Il2rγnull mutation, predominantly NOD.Cg-Prkdcscid Il2rγtm1Wjl/SzJ (NSG) mice. The diarrhea was transmissible to naïve NSG mice by co-housing or gavage of intestinal homogenates from diarrheic mice, suggesting the involvement of an infectious agent and thus was given the name "infectious diarrhea." Conventional testing failed to identify an etiology. Strain-resolved metagenomic analyses using DNA from diarrheic and control fecal samples yielded the genome sequence of an enterotoxin-encoding Clostridium cuniculi strain, a candidate factor underlying the diarrhea outbreak. We hypothesized that the presence of C. cuniculi and its enterotoxin in fecal samples could serve as biomarkers. Quantitative real-time PCR (qPCR) assays using specific primers for C. cuniculi and its enterotoxin were generated and validated. We analyzed fecal samples from 111 NSG or NSG-related mice that were healthy, 37 that had clinical signs of infectious diarrhea, and 28 that had diarrhea attributable to known causes. Positive qPCR results for C. cuniculi and its enterotoxin only occurred in feces from mice with infectious diarrhea. All positive samples contained both C. cuniculi and its enterotoxin. Our data suggest that infectious diarrhea in these cases is mediated, at least in part, by the transmission of C. cuniculi and its enterotoxin. Our novel qPCR assays for C. cuniculi and its enterotoxin are effective tools for the detection of infectious diarrhea in NSG mice.

  View details for DOI 10.1177/10406387251378688

  View details for PubMedID 40974247

• Clostridium cuniculi is associated with chronic high-morbidity low-mortality diarrhea in NSG and NSG-related mouse strains. Veterinary pathology Casey, K. M., Barouch-Bentov, R., Reineking, W., Alonso, F. H., Moorhead, R., Fazel, M., Armien, A. G., Uzal, F. A., Chanin, R. B., Bhatt, A. S., Green, S. L., Felt, S. A., Nagamine, C. M. 2025: 3009858251372565

  Abstract

  In October 2020, adult male and female NSG (NOD. Cg-Prkdcscid Il2rgtm1Wjl/Sz) mice were reported for diarrhea within a mouse barrier facility. Other immunodeficient strains harboring the SCID (Prkdcscid) or Rag (Ragnull) mutations together with the IL2rg (Il2rgnull) mutation were affected. At its peak, over 20 laboratories in 10/16 (62.5%) barrier rooms were affected. Mortality was rare except in lactating females (≥ P11). Grossly, nonlactating adult female and male mice (n = 16) had mild to moderate, small and large intestinal distension with corresponding individual cell death and sloughing of superficial enterocytes in the cecocolonic mucosa. Lactating NSG dams (n=6) had moderate to severe gastrointestinal distension and/or segmental, dark red to gray, small intestinal discoloration. In addition to the same histologic lesions seen in nonlactating female NSG mice, lactating NSG dams often had severe ulcerative inflammation affecting the jejunum, ileum, cecum, and colon. Traditional ancillary diagnostic tests including aerobic and anaerobic cultures (blood, liver, spleen, and intestines), fecal PCR, and fecal floatation failed to yield a causative organism. Further cohousing and oral gavage studies determined neither immunocompetent CD1 (Crl:CD1 [ICR]) mice nor immunodeficient NOD scid (NOD.Cg-Prkdcscid/J) and Rag2 KO (C57BL/6. Cg-Rag2tm1.1Cgn/J) mice were susceptible to clinical disease. Extensive control barriers were implemented including a veterinary-managed NSG breeding barrier, alterations in husbandry practices, and strategic environmental disinfection, allowing for continuity of experimental studies while avoiding widespread depopulation of the barrier. Subsequent strain-resolved metagenomics and qPCR assay development identified Clostridium cuniculi and its enterotoxin exclusively within diarrheic mice.

  View details for DOI 10.1177/03009858251372565

  View details for PubMedID 40974275

• Color-neutral and reversible tissue transparency enables longitudinal deep-tissue imaging in live mice. Proceedings of the National Academy of Sciences of the United States of America Keck, C. H., Schmidt, E. L., Roth, R. H., Floyd, B. M., Tsai, A. P., Garcia, H. B., Cui, M., Chen, X., Wang, C., Park, A., Zhao, S., Liao, P. A., Casey, K. M., Reineking, W., Cai, S., Zhang, L. Y., Yang, Q., Yuan, L., Baghdasaryan, A., Lopez, E. R., Cooper, L., Cui, H., Esquivel, D., Brinson, K., Chen, X., Wyss-Coray, T., Coleman, T. P., Brongersma, M. L., Bertozzi, C. R., Wang, G. X., Ding, J. B., Hong, G. 2025; 122 (35): e2504264122

  Abstract

  Light scattering in biological tissue presents a significant challenge for deep in vivo imaging. Our previous work demonstrated the ability to achieve optical transparency in live mice using intensely absorbing dye molecules, which created transparency in the red spectrum while blocking shorter-wavelength photons. In this paper, we extend this capability to achieve optical transparency across the entire visible spectrum by employing molecules with strong absorption in the ultraviolet spectrum and sharp absorption edges that rapidly decline upon entering the visible spectrum. This color-neutral and reversible tissue transparency method enables optical transparency for imaging commonly used fluorophores in the green and yellow spectra. Notably, this approach facilitates tissue transparency for structural and functional imaging of the live mouse brain labeled with yellow fluorescent protein and GCaMP through the scalp and skull. We show that this method enables longitudinal imaging of the same brain regions in awake mice over multiple days during development. Histological analyses of the skin and systemic toxicology studies indicate minimal acute or chronic damage to the skin or body using this approach. This color-neutral and reversible tissue transparency technique opens opportunities for noninvasive deep-tissue optical imaging, enabling long-term visualization of cellular structures and dynamic activity with high spatiotemporal resolution and chronic tracking capabilities.

  View details for DOI 10.1073/pnas.2504264122

  View details for PubMedID 40857313

• Reply to: "EFNB3 Frameshift Variant in Weimaraner Dogs with a Condition Resembling a Congenital Mirror Movement Disorder". Movement disorders : official journal of the Movement Disorder Society Schwarz, C., Bartenschlager, F., Kershaw, O., Braun, J., Guevar, J., Jagannathan, V., Epplen, J. T., Reineking, W., Baumgartner, W., Bhatia, K. P., Gruber, A. D., Leeb, T. 2025

  View details for DOI 10.1002/mds.70000

  View details for PubMedID 40772476

• Analysis of knockout mice reveals critical female-specific roles for the Hippo pathway component PTPN14. Genes & development McCrea, E. M., Makrides, N., Tabata, T., Reineking, W., Vilches-Moure, J. G., Wang, M., Lake, J. S., Vogel, H., Howitt, B., Zhang, X., Attardi, L. D. 2025

  Abstract

  The Hippo pathway regulates many physiological processes, including development, tumor suppression, and wound healing. One understudied Hippo pathway component is PTPN14, an evolutionarily conserved tyrosine phosphatase that inhibits YAP/TAZ. Although it is an established tumor suppressor, PTPN14's role in tissue homeostasis has remained unclear. We thus generated Ptpn14-deficient mice and found that only ∼60% of Ptpn14 -/- mice survived postnatally, highlighting the importance of PTPN14 for viability while also enabling the discovery of PTPN14 physiological functions. Ptpn14 -/- mice developed debilitating corneal lesions and the uterus defect hydrometra, as well as heart and kidney abnormalities. Ptpn14 deficiency precipitated an impaired injury response in the cornea and dysregulated YAP signaling in both the uterus and the cornea. Notably, these phenotypes were female-specific, revealing sexually dimorphic Hippo pathway function through PTPN14. Finally, analysis of human PTPN14 variants suggested that PTPN14's essential roles are conserved in humans, underscoring the importance of our insights for designing therapies to improve women's health.

  View details for DOI 10.1101/gad.352620.125

  View details for PubMedID 40533389

• Synergistic interference with SARS-CoV-2 replication by molnupiravir-derived N4-hydroxycytidine and inhibitors of CTP synthetase in cell culture. Virology Stegmann, K. M., Dickmanns, A., Fuchs, H. L., Scheibner, D., Mohl, B. P., Moeselaken, F. R., Reineking, W., Störk, T., Volz, A., Beer, P. A., Parker, A. E., Pilchova, V., Meyer Zu Natrup, C., von Köckritz-Blickwede, M., Baumgärtner, W., Balkema-Buschmann, A., Dobbelstein, M. 2025; 610: 110598

  Abstract

  N4-hydroxycytidine (NHC), the active metabolite of molnupiravir, is incorporated into nascent RNA of SARS-CoV-2 and interferes with subsequent virus replication. We have previously described synergy between NHC and inhibitors of dehydroorotate dehydrogenase (DHODH), an enzyme required for pyrimidine synthesis. Upon DHODH inhibition, the lack of endogenous pyrimidines conceivably enhances NHC incorporation. However, the question remains whether preventing the synthesis of just one pyrimidine base, cytidine, might as well augment the antiviral efficacy of NHC. We tested this by inhibiting CTP synthetases (CTPSs), the cellular enzymes that directly catalyze the synthesis of a cytidine nucleotide. We observed that inhibitors of CTP synthetase (CTPSis), namely cyclopentenyl cytosine (CPEC) as well as STP938 and STP720, display a strong synergy with NHC for diminishing SARS-CoV-2 replication in cell culture, as shown earlier for DHODH inhibitors. NHC and CTPSis in combination prevented the cytopathic effect of SARS-CoV-2 and strongly reduced the release of viral RNA and infectious particles, as well as the synthesis of viral proteins. This combination was also active against an Omicron variant of SARS-CoV-2. Addition of cytidine, but not uridine, rescued virus growth under these conditions. Surprisingly, this synergy was not confirmed in the SARS-CoV-2 animal model in Syrian hamsters. While treatment with the CTPS1 inhibitor STP938 alone strongly diminished virus propagation and COVID pathology, addition of molnupiravir did not augment this effect and even counteracted the benefits of STP938 in vivo. We propose that, if further developed, CTPS inhibitors might represent candidates for antiviral therapy.

  View details for DOI 10.1016/j.virol.2025.110598

  View details for PubMedID 40561865

• <i>EFNB3</i> Frameshift Variant in Weimaraner Dogs with a Condition Resembling a Congenital Mirror Movement Disorder MOVEMENT DISORDERS Schwarz, C., Bartenschlager, F., Kershaw, O., Braun, J., Guevar, J., Jagannathan, V., Epplen, J. T., Reineking, W., Baumgaertner, W., Bhatia, K. P., Gruber, A. D., Leeb, T. 2025

  Abstract

  Congenital mirror movement disorders (CMMs) are clinically and genetically heterogeneous in human patients. CMMs have not been documented to occur spontaneously in animals.The objective of this work was to document the first case of CMMs spontaneously occurring in Weimaraner dogs and to identify the underlying genetic cause.Clinical and pathological investigations were performed. Genetic investigations used linkage and autozygosity mapping followed by whole-genome sequencing of 3 affected dogs and 1489 control dogs to identify disease-associated variants.Three of 11 puppies in a litter of Weimaraner dogs exhibited an abnormal gait characterized by synchronized saltatorial locomotion. Their phenotype was tentatively termed congenital mirror movement disorder 1 (CMM1). The underlying genetic cause was identified as a 2-bp duplication in EFNB3 encoding ephrin-B3, a transmembrane protein important for axon guidance and spinal midline barrier formation during neurodevelopment. The identified variant, XM_038536724.1:c.643_644dup, is predicted to lead to a frameshift and introduction of a premature stop codon XP_038392652.1:p.(Ala216Valfs*79). CMM1 is inherited as an autosomal recessive trait in these dogs.Similar to humans, CMMs may occur in dogs as an inherited disease as a result of a spontaneously arisen genetic variant. The CMM1 phenotype in dogs resembles the phenotype of experimentally induced Efnb3-/- knockout mice. So far, no human patients with EFNB3-related CMMs have been reported. Our study provides the first naturally occurring large-animal model for CMMs. EFNB3 should be considered a candidate gene in human CMM patients with unclear disease etiology. © 2025 International Parkinson and Movement Disorder Society.

  View details for DOI 10.1002/mds.30243

  View details for Web of Science ID 001492426700001

  View details for PubMedID 40401490

• Somatic hypermutation shapes the viral escape profile of SARS-CoV-2 neutralising antibodies. EBioMedicine Bruhn, M., Obara, M., Gonzalez-Hernandez, M., Reineking, W., Salam, A., Mirolo, M., Hinrichs, I., Mergani, A., Bartsch, Y., Schambach, A., Zimmer, G., Baumgartner, W., Osterhaus, A. D., Kalinke, U. 2025; 116: 105770

  Abstract

  BACKGROUND: Since the onset of the COVID-19 pandemic, SARS-CoV-2 neutralising monoclonal antibodies (mAbs) are being developed for clinical use. With the appearance of new virus variants, most mAbs lost their virus-neutralising activity, highlighting the complexity of mAb development under conditions of continuous SARS-CoV-2 evolution.METHODS: Hamsters were treated with SARS-CoV-2 neutralising mAbs and then challenged with SARS-CoV-2. Recombinant VSV expressing the spike protein of SARS-CoV-2 was utilised in an in vitro system to select for antibody escape variants. Surface plasmon resonance measurements were performed to characterise the binding affinity and epitope of various mAbs. Fc-mediated effector functions of neutralising and non-neutralising mAb combinations were determined via multiple in vitro assays.FINDINGS: Few of the mAb treated and infected hamsters experienced breakthrough infections, which derived from mutated virus that emerged in vivo. We developed an in vitro antibody escape assay that recapitulated the in vivo situation and we found that somatic hypermutations (SHM) affected the profile of viral escape hotspots that mAbs selected for. Pairwise combination of mAbs binding non-overlapping epitopes suppressed the emergence of viral mutants. The formulation with a third, non-neutralising mAb enhanced the Fc-mediated effector functions of the mAb treatment in an additive manner.INTERPRETATION: We conclude that treatment with single mAbs rapidly leads to the formation of novel virus variants. An important function of SHM is to suppress the emergence of viral antibody escape variants. Our data suggest that the anticipatory B cell memory can be harnessed to design combinations of SARS-CoV-2 neutralising mAbs that have a reduced risk to induce viral escape.FUNDING: This study was supported by public funding from the German Research Foundation (DFG), the Federal Ministry of Education and Research (BMBF), the COVID-19-Research Network of the State of Lower Saxony (COFONI), the German Centre for Infection Research (DZIF), and the Helmholtz Association of German Research Centres.

  View details for DOI 10.1016/j.ebiom.2025.105770

  View details for PubMedID 40403696

• Respiratory long COVID in aged hamsters features impaired lung function post-exercise with bronchiolization and fibrosis NATURE COMMUNICATIONS Heydemann, L., Ciurkiewicz, M., Stoerk, T., Zdora, I., Huelskoetter, K., Gregor, K., Michaely, L., Reineking, W., Schreiner, T., Beythien, G., Volz, A., Tuchel, T., Meyer zu Natrup, C., Schuenemann, L., Clever, S., Henneck, T., von Koeckritz-Blickwede, M., Schaudien, D., Rohn, K., Schughart, K., Geffers, R., Kaneko, M. K., Kato, Y., Gross, C., Amanakis, G., Pavlou, A., Baumgaertner, W., Armando, F. 2025; 16 (1): 2080

  Abstract

  Long-term consequences of SARS-CoV-2 infection affect millions of people and strain public health systems. The underlying pathomechanisms remain unclear, necessitating further research in appropriate animal models. This study aimed to characterize the trajectory of lung regeneration over 112 days in the male hamster model by combining morphological, transcriptomic and functional readouts. We demonstrate that in the acute phase, SARS-CoV-2 Delta-infected, male, aged hamsters show a severe impairment of lung function at rest. In the chronic phase, similar impairments persisted up to 7 weeks post-infection but were only evident after exercise on a rodent treadmill. The male hamster model recapitulates chronic pulmonary fibrotic changes observed in many patients with respiratory long COVID, but lacks extra-pulmonary long-term lesions. We show that sub-pleural and interstitial pulmonary fibrosis as well as alveolar bronchiolization persist until 112 dpi. Interestingly, CK8+ alveolar differentiation intermediate (ADI) cells are becoming less prominent in the alveolar proliferation areas from 28 dpi on. Instead, CK14+ airway basal cells and SCGB1A1+ club cells, expressing cell proliferation markers, mainly populate alveolar bronchiolization areas at later time-points. We postulate that pulmonary fibrosis and SCGB1A1+ club cell-rich areas of alveolar bronchiolization represent potential risk factors for other diseases in long-COVID survivors.

  View details for DOI 10.1038/s41467-025-57267-x

  View details for Web of Science ID 001435592900035

  View details for PubMedID 40021627

  View details for PubMedCentralID PMC11871369

• Color-neutral and reversible tissue transparency enables longitudinal deep-tissue imaging in live mice. bioRxiv : the preprint server for biology Keck, C. H., Schmidt, E. L., Roth, R. H., Floyd, B. M., Tsai, A. P., Garcia, H. B., Cui, M., Chen, X., Wang, C., Park, A., Zhao, S., Liao, P. A., Casey, K. M., Reineking, W., Cai, S., Zhang, L., Yang, Q., Yuan, L., Baghdasaryan, A., Lopez, E. R., Cooper, L., Cui, H., Esquivel, D., Brinson, K., Chen, X., Wyss-Coray, T., Coleman, T. P., Brongersma, M. L., Bertozzi, C. R., Wang, G. X., Ding, J. B., Hong, G. 2025

  Abstract

  Light scattering in biological tissue presents a significant challenge for deep in vivo imaging. Our previous work demonstrated the ability to achieve optical transparency in live mice using intensely absorbing dye molecules, which created transparency in the red spectrum while blocking shorter-wavelength photons. In this paper, we extend this capability to achieve optical transparency across the entire visible spectrum by employing molecules with strong absorption in the ultraviolet spectrum and sharp absorption edges that rapidly decline upon entering the visible spectrum. This new color-neutral and reversible tissue transparency method enables optical transparency for imaging commonly used fluorophores in the green and yellow spectra. Notably, this approach facilitates tissue transparency for structural and functional imaging of the live mouse brain labeled with yellow fluorescent protein and GCaMP through the scalp and skull. We show that this method enables longitudinal imaging of the same brain regions in awake mice over multiple days during development. Histological analyses of the skin and systemic toxicology studies indicate minimal acute or chronic damage to the skin or body using this approach. This color-neutral and reversible tissue transparency technique opens new opportunities for noninvasive deep-tissue optical imaging, enabling long-term visualization of cellular structures and dynamic activity with high spatiotemporal resolution and chronic tracking capabilities.

  View details for DOI 10.1101/2025.02.20.639185

  View details for PubMedID 40060493

• Syrian hamsters (Mesocricetus auratus) as an upper respiratory tract model for respiratory syncytial virus infection. Npj viruses Kolbe, S. M., Guilfoyle, K., Reineking, W., van Amerongen, G., van der Net, G., Lockow, S., Baumgärtner, W., Ludlow, M., Osterhaus, A. D. 2025; 3 (1): 2

  Abstract

  Respiratory syncytial virus (RSV) is one of the leading causes of respiratory tract infection in children, immunocompromised individuals and older adults. Vaccines have recently been approved for use in adults and although further efforts to develop suitable interventions for children are ongoing, there are limited animal models for RSV infection. For preclinical efficacy testing of prophylactic and therapeutic treatments cotton rat and ferret models can be used. However, these can be expensive, difficult to source and house, and often have limitations such as insufficient virus replication in the respiratory tract and/or lack of horizontal transmission. In this study, Syrian hamsters (Mesocricetus auratus), which are relatively cheap, easy to source and house, were inoculated intranasally with a recombinant RSV-A-0594 strain expressing EGFP and using virological and pathological analyses. Viral replication was assessed and compared to viral replication in the ferret model. Although there was limited virus infection of the lower respiratory tract of Syrian hamsters, we show that a contemporary recombinant RSV-A strain replicates efficiently in the upper respiratory tract of Syrian hamsters (titers up to 4.5 Log10 TCID50/g and 12 Log10 RNA copies/g). These titers are comparable to those found in the ferret upper respiratory tract tissues post-infection with the same virus strain (up to 6.0 Log10 TCID50/g and 12 Log 10 RNA copies/g). Fluorescent regions indicating virus infection were macroscopically visible under UV-light in the nasal turbinates and histological assessment showed mucosal inflammation with necrotic cells in this tissue. In summary, Syrian hamsters generally displayed less severe systemic and pulmonary changes than ferrets, but do appear to be a promising model for upper respiratory tract infection with RSV.

  View details for DOI 10.1038/s44298-024-00086-6

  View details for PubMedID 40295717

  View details for PubMedCentralID PMC11721388

• Inhibitors of dihydroorotate dehydrogenase synergize with the broad antiviral activity of 4'-fluorouridine. Antiviral research Schrell, L., Fuchs, H. L., Dickmanns, A., Scheibner, D., Olejnik, J., Hume, A. J., Reineking, W., Stork, T., Muller, M., Graaf-Rau, A., Diederich, S., Finke, S., Baumgartner, W., Muhlberger, E., Balkema-Buschmann, A., Dobbelstein, M. 2024: 106046

  Abstract

  RNA viruses present a constant threat to human health, often with limited options for vaccination or therapy. Notable examples include influenza viruses and coronaviruses, which have pandemic potential. Filo- and henipaviruses cause more limited outbreaks, but with high case fatality rates. All RNA viruses rely on the activity of a virus-encoded RNA-dependent RNA polymerase (RdRp). An antiviral nucleoside analogue, 4'-Fluorouridine (4'-FlU), targets RdRp and diminishes the replication of several RNA viruses, including influenza A virus and SARS-CoV-2, through incorporation into nascent viral RNA and delayed chain termination. However, the effective concentration of 4'-FlU varied among different viruses, raising the need to fortify its efficacy. Here we show that inhibitors of dihydroorotate dehydrogenase (DHODH), an enzyme essential for pyrimidine biosynthesis, can synergistically enhance the antiviral effect of 4'-FlU against influenza A viruses, SARS-CoV-2, henipaviruses, and Ebola virus. Even 4'-FlU-resistant mutant influenza A virus was re-sensitized towards 4'-FlU by DHODH inhibition. The addition of uridine rescued influenza A virus replication, strongly suggesting uridine depletion as a mechanism of this synergy. 4'-FlU was also highly effective against SARS-CoV-2 in a hamster model of COVID. We propose that the impairment of endogenous uridine synthesis by DHODH inhibition enhances the incorporation of 4'-FlU into viral RNAs. This strategy may be broadly applicable to enhance the efficacy of pyrimidine nucleoside analogues for antiviral therapy.

  View details for DOI 10.1016/j.antiviral.2024.106046

  View details for PubMedID 39638153

• Ex vivo comparison of full-thickness biopsy techniques in the equine small intestine. Veterinary surgery : VS Verhaar, N., Hammer, E., Reineking, W., Hewicker-Trautwein, M., Geburek, F. 2024

  Abstract

  To compare the practicability and tissue sample quality between different intestinal biopsy techniques.Experimental, randomized ex vivo study.Small intestine of nine horses.Four different biopsy techniques were evaluated in the aboral jejunum and the ileum within 1 h after euthanasia. One segment was used as control (C), and the applied techniques included an 8 mm biopsy punch (BP), transverse wedge resection (TW), longitudinal wedge resection with transverse closure (LW) and a longitudinal sample using Eppendorfer biopsy forceps (EF). Defects were closed using a single-layer continuous Lembert pattern. Duration of the procedure, intestinal diameter, contamination, and bursting pressure were determined. The quality of the obtained tissue samples for histological assessment was evaluated using a semiquantitative score. The jejunal and ileal samples were analyzed separately.All biopsy procedures including defect closure were completed within 5 min, with shorter closure times for BP (p = .03). Minimal contamination could be noted in 1/8 TW and 2/8 LW cases, without significant differences between the groups. Longitudinal closure techniques (BP, EF) showed more constriction than transverse closures (TW, LW) (p < .05). Bursting pressure was >75 mmHg in all cases. Technique BP showed significantly lower biopsy quality scores (p = .009).The tested biopsy techniques could all be applied effectively within a reasonable time frame, yet the biopsy punch was associated with significant artifacts and risk of missing mucosa.The findings provide insights into the possible advantages and limitations of the different techniques and alert the surgeon to potential issues with the quality of the tissue sample.

  View details for DOI 10.1111/vsu.14178

  View details for PubMedID 39404177

• Flowmetry and spectrophotometry can detect reduced intestinal microperfusion in nonsurvivors during equine colic surgery for large intestinal strangulation. American journal of veterinary research Verhaar, N., Reineking, W., Hewicker-Trautwein, M., Grages, A. M., Kästner, S. B., Geburek, F. 2024: 1-8

  Abstract

  To evaluate the use of laser Doppler flowmetry and spectrophotometry (LDFS) for large intestinal viability assessment in horses with naturally occurring large intestinal strangulations.By use of LDFS, intestinal microperfusion was quantified as tissue oxygen saturation (tSo2), hemoglobin (tHB), and blood flow (tBF) in cases with large colon volvulus and small colon strangulations undergoing colic surgery (n = 17). Intestinal biopsies were taken from the pelvic flexure in all large colon cases and in small colon cases that underwent intraoperative euthanasia. Measurements were compared between survivors and nonsurvivors, and the correlation between LDFS and (immuno)histology was tested (P < .05).The tSo2 and tBF were clearly lower and tHB was higher than previously reported in healthy horses. Following correction of the lesion, pelvic flexure tBF was significantly lower than that of the left ventral colon. Prior to correction of the lesion, microperfusion did not differ between survivors and nonsurvivors, but following release of the strangulation the survivors had a significantly higher tSo2 and tBF compared to the nonsurvivors. There was a negative correlation between tBF and interstitium-to-crypt ratio and a positive correlation between tHB and the histological hemorrhage score. There were no significant correlations between LDFS measurements and inflammatory cell counts or hypoxia-inducible factor-1α immunoreactivity.Large intestinal microperfusion was decreased in nonsurvivors compared to survivors and was correlated with histological injury, suggesting that LDFS has the potential to predict tissue injury and postoperative survival.The use of LDFS as an ancillary diagnostic aid may improve intraoperative viability assessment during colic surgery.

  View details for DOI 10.2460/ajvr.24.05.0142

  View details for PubMedID 39116909

• Successful Treatment of an Acinar Pancreatic Carcinoma in an Inland Bearded Dragon (Pogona vitticeps): A Case Report. Animals : an open access journal from MDPI Hetterich, J., Hewicker-Trautwein, M., Reineking, W., Allnoch, L., Pees, M. 2024; 14 (13)

  Abstract

  An adult, 362 g, male, intact inland bearded dragon (Pogona vitticeps) was admitted to a veterinary clinic due to a temporary cloacal prolapse and a two-week history of reduced overall condition and forage intake. Physical examination revealed an approximately 2 × 1 cm round-shaped, rigid intracoelomic tissue mass. Multiple sand deposits were present on the cloacal mucous membranes, though no signs of cloacal prolapse were present. The lizard was otherwise responsive but showed reduced body tension and movement behavior. Initial fecal examination revealed a high-grade oxyuriasis. A 2 × 1.5 cm sized intracoelomic, well-vascularized, round-shaped mass was subsequently visualized by ultrasonography. After a two-day stabilization therapy, the intracoelomic mass was removed by performing a standard ventral coeliotomy under general anesthesia. Histopathological examination of the excised mass revealed an acinar pancreatic adenocarcinoma with infiltration of the peritumorous connective soft tissue. The lizard remained at the clinic for a further seven days. Its postsurgical condition improved slowly. However, the lizard started regular forage intake 10 days after surgery, and general behavior enhanced constantly within the following three weeks. The animal was presented for a follow-up six weeks after surgery, showing bright and alert behavior with no signs of disease or illness. The lizard was re-examined 20 months after the initial presentation due to a reduced overall condition and reduced food intake. Blood chemistry evaluation revealed markedly decreased protein parameters, and moderate ascites was identified ultrasonographically. A distinct association with the preceding neoplastic disease could not be made, and the lizard returned to its regular condition under supportive therapy within three weeks. To the authors' knowledge, this is the first report of successful treatment of a pancreatic carcinoma in a bearded dragon.

  View details for DOI 10.3390/ani14131976

  View details for PubMedID 38998088

• Flowmetry and spectrophotometry for the assessment of intestinal viability in horses with naturally occurring strangulating small intestinal lesions. Equine veterinary journal Verhaar, N., Grages, A. M., Bienert-Zeit, A., Schwieder, A., Reineking, W., Hewicker-Trautwein, M., Kästner, S., Geburek, F. 2024

  Abstract

  Ancillary diagnostic methods to enhance the accuracy of viability assessment have not been established for use in clinical practice.To assess intestinal microperfusion measured by Laser Doppler Flowmetry and Spectrophotometry (LDFS) in naturally occurring small intestinal strangulations of different origins and to compare this between viable and non-viable segments.Prospective clinical trial.Forty horses undergoing colic surgery for naturally occurring small intestinal strangulations were included. Tissue oxygen saturation (tSO2), haemoglobin (tHB) and blood flow (tBF) were determined by LDFS before and after release of the strangulation. Intestinal biopsies were taken in cases that underwent intestinal resection or intraoperative euthanasia and assessed using a semi-quantitative mucosal injury score (MIS). The LDFS measurements were compared between the different categories of strangulation causes and histopathological injury using parametric and non-parametric tests (p < 0.05).Strangulations by pedunculated lipomas had lower tBF (13.9 ± 18 arbitrary units [AU]) than epiploic foramen entrapments (65.2 ± 61 AU; CI -1.697 to -0.2498; p = 0.005). Segments with MIS > 5 showed lower tBF during strangulation than segments with MIS < 4 (mean difference 61.1 AU; CI -1.119 to -0.07361; p = 0.03). This did not differ significantly following release of strangulation. Furthermore, there was a positive correlation between the inflammatory cell count and tBF during strangulation (r 0.34; CI 0.01 to 0.60; p = 0.04). The tSO2 and tHB did not differ between the different categories of lesions or injury.No biopsies could be taken from the intestinal segments that did not undergo resection. The duration of strangulation could not reliably be ascertained.Blood flow measurements in naturally occurring strangulating lesions show a varying degree of ischaemia in different causes of strangulation. Intestinal blood flow measurements prior to release of the strangulation could potentially contribute to the identification of mucosal injury, yet a high individual variability and other contributing factors need to be considered.

  View details for DOI 10.1111/evj.14118

  View details for PubMedID 38888520

• Spontaneous Lethal Outbreak of Influenza A Virus Infection in Vaccinated Sows on Two Farms Suggesting the Occurrence of Vaccine-Associated Enhanced Respiratory Disease with Eosinophilic Lung Pathology. Viruses Reineking, W., Hennig-Pauka, I., Schröder, L., Höner, U., Schreiber, E., Geiping, L., Lassnig, S., Bonilla, M. C., Hewicker-Trautwein, M., de Buhr, N. 2024; 16 (6)

  Abstract

  Influenza A virus (IAV) infections in swine are usually subclinical, but they can reach high morbidity rates. The mortality rate is normally low. In this study, six vaccinated, spontaneously deceased sows revealed IAV infection and enhanced neutrophilic bronchopneumonia with unexpectedly large numbers of infiltrating eosinophils. The purpose of this study was to characterize these lung lesions with special emphasis on the phenotypes of inflammatory cells, the presence of eosinophilic peroxidase (EPO), and neutrophil extracellular traps (NETs). The number of Sirius red-stained eosinophils was significantly higher in the lungs of IAV-infected sows compared to healthy pigs, indicating a migration of eosinophils from blood vessels into the lung tissue stimulated by IAV infection. The detection of intra- and extracellular EPO in the lungs suggests its contribution to pulmonary damage. The presence of CD3+ T lymphocytes, CD20+ B lymphocytes, and Iba-1+ macrophages indicates the involvement of cell-mediated immune responses in disease progression. Furthermore, high numbers of myeloperoxidase-positive cells were detected. However, DNA-histone-1 complexes were reduced in IAV-infected sows, leading to the hypothesis that NETs are not formed in the IAV-infected sows. In conclusion, our findings in the lungs of IAV-infected vaccinated sows suggest the presence of so far unreported field cases of vaccine-associated enhanced respiratory disease.

  View details for DOI 10.3390/v16060955

  View details for PubMedID 38932247

  View details for PubMedCentralID PMC11209110

• Melan-A immunolabeling in canine extramedullary plasmacytomas. Veterinary pathology Schuwerk, L., Ulianytska, A., Baumgärtner, W., Reineking, W. 2024: 3009858241246979

  Abstract

  Histologic diagnosis of less well-differentiated cases of canine extramedullary plasmacytomas (CEMPs) may require immunohistochemical confirmation to discriminate these tumors from other round cells tumors including lymphoma, cutaneous histiocytoma, and amelanotic melanomas. CEMPs are characterized by widespread immunoreactivity for multiple myeloma 1 (MUM1) antigen and λ light chains, while the melanocytic marker melan-A has been reported to yield negative results. Here, 33 randomly selected CEMPs, 20 melanocytomas, and 20 malignant melanomas were immunohistochemically tested for MUM1, melan-A, and PNL2. In addition, CEMPs were examined for PAX5, E-cadherin, CD3, CD18, CD20, S100, as well as λ and κ light chain immunoreactivity. All CEMPs were characterized by labeling for MUM1 and λ light chain, as well as variable immunopositivity for the remaining antibodies. Notably, 13 cases of CEMPs (39.4%) exhibited immunolabeling for melan-A. Melanocytic tumors immunolabeled for melan-A (40/40; 100%) and PNL2 (34/40; 85%). An unexpected cytoplasmic immunoreactivity for MUM1 was observed in 2 melanocytic tumors. Summarized, MUM1 or melan-A immunomarkers alone are not sufficient to differentiate between CEMPs and amelanotic melanomas and should be part of a larger immunopanel including λ light chain, CD20, and PNL2.

  View details for DOI 10.1177/03009858241246979

  View details for PubMedID 38642035

• Measuring tissue oxygen saturation in the orad intestinal segment during equine colic surgery may aid in predicting the occurrence of postoperative ileus. American journal of veterinary research Verhaar, N., Grages, A. M., Sauer, F. J., Geiger, T., Reineking, W., Hewicker-Trautwein, M., Geburek, F., Kästner, S. B. 2024: 1-8

  Abstract

  To assess the histological injury and intestinal microperfusion measured by laser Doppler flowmetry and spectrophotometry (LDFS) of the small intestine orad to a strangulation during colic surgery.Horses with naturally occurring small intestinal strangulations undergoing colic surgery were included.In this prospective clinical trial, intestinal tissue oxygen saturation (tSO2) and tissue blood flow (tBF) were measured by LDFS orad to the strangulation following release of the strangulation (n = 18). The number of horses with postoperative reflux (POR) and the cases that survived until discharge were compared between groups using Fisher's exact test (P < .05). Intestinal biopsies were taken in cases that underwent intestinal resection or intraoperative euthanasia (n = 28). Measurements were compared between injured and noninjured segments with a Mann-Whitney U or t test.The tSO2 and tBF of the orad intestine were lower than previously reported in healthy horses. Horses with low tSO2 of < 35% were significantly more likely to suffer from POR (6/6 cases) compared to cases with tSO2 > 69% (1/6). The number of horses that survived were not statistically different between these groups (2/6 and 6/6). All horses with mucosal injury developed POR (6/6), which was significantly more likely compared to horses without mucosal injury (3/13). No significant difference in tSO2 or tBF could be found between the segments with and without histological injury.The results suggest that measuring tSO2 in the orad segment during colic surgery may aid in predicting postoperative issues.

  View details for DOI 10.2460/ajvr.23.12.0286

  View details for PubMedID 38626792

• Concurrent Detection of a Papillomatous Lesion and Sequence Reads Corresponding to a Member of the FamilyAdintoviridaein a Bell's Hinge-Back Tortoise (Kinixys belliana). Animals : an open access journal from MDPI Hetterich, J., Mirolo, M., Kaiser, F., Ludlow, M., Reineking, W., Zdora, I., Hewicker-Trautwein, M., Osterhaus, A. D., Pees, M. 2024; 14 (2)

  Abstract

  An adult male Bell's hinge-back tortoise (Kinixys belliana) was admitted to a veterinary clinic due to a swelling in the oral cavity. Physical examination revealed an approximately 2.5 * 1.5 cm sized, irregularly shaped tissue mass with villiform projections extending from its surface located in the oropharyngeal cavity. An initial biopsy was performed, and the lesion was diagnosed as squamous papilloma. Swabs taken for virological examination tested negative with specific PCRs for papillomavirus and herpesvirus. Further analysis of the oropharyngeal mass via metagenomic sequencing revealed sequence reads corresponding to a member of the family Adintoviridae. The tissue mass was removed one week after the initial examination. The oral cavity remained unsuspicious in follow-up examinations performed after one, five and twenty weeks. However, a regrowth of the tissue was determined 23 months after the initial presentation. The resampled biopsy tested negative for sequence reads of Adintoviridae. Conclusively, this report presents the diagnostic testing and therapy of an oral cavity lesion of unknown origin. The significance of concurrent metagenomic determination of adintovirus sequence reads within the tissue lesion is discussed.

  View details for DOI 10.3390/ani14020247

  View details for PubMedID 38254416

• Nanobodies to multiple spike variants and inhalation of nanobody-containing aerosols neutralize SARS-CoV-2 in cell culture and hamsters. Antiviral research Aksu, M., Kumar, P., Güttler, T., Taxer, W., Gregor, K., Mußil, B., Rymarenko, O., Stegmann, K. M., Dickmanns, A., Gerber, S., Reineking, W., Schulz, C., Henneck, T., Mohamed, A., Pohlmann, G., Ramazanoglu, M., Mese, K., Groß, U., Ben-Yedidia, T., Ovadia, O., Fischer, D. W., Kamensky, M., Reichman, A., Baumgärtner, W., von Köckritz-Blickwede, M., Dobbelstein, M., Görlich, D. 2023; 221: 105778

  Abstract

  The ongoing threat of COVID-19 has highlighted the need for effective prophylaxis and convenient therapies, especially for outpatient settings. We have previously developed highly potent single-domain (VHH) antibodies, also known as nanobodies, that target the Receptor Binding Domain (RBD) of the SARS-CoV-2 Spike protein and neutralize the Wuhan strain of the virus. In this study, we present a new generation of anti-RBD nanobodies with superior properties. The primary representative of this group, Re32D03, neutralizes Alpha to Delta as well as Omicron BA.2.75; other members neutralize, in addition, Omicron BA.1, BA.2, BA.4/5, and XBB.1. Crystal structures of RBD-nanobody complexes reveal how ACE2-binding is blocked and also explain the nanobodies' tolerance to immune escape mutations. Through the cryo-EM structure of the Ma16B06-BA.1 Spike complex, we demonstrated how a single nanobody molecule can neutralize a trimeric spike. We also describe a method for large-scale production of these nanobodies in Pichia pastoris, and for formulating them into aerosols. Exposing hamsters to these aerosols, before or even 24 h after infection with SARS-CoV-2, significantly reduced virus load, weight loss and pathogenicity. These results show the potential of aerosolized nanobodies for prophylaxis and therapy of coronavirus infections.

  View details for DOI 10.1016/j.antiviral.2023.105778

  View details for PubMedID 38065245

• Histological and immunohistochemical characterization of mucosa-associated lymphoid tissue and antigen-presenting cells in trachea and lung of cattle ANATOMIA HISTOLOGIA EMBRYOLOGIA Thomasmeyer, A., Reineking, W., Hewicker-Trautwein, M. 2023

  Abstract

  The presence of bronchus-associated lymphoid tissue (BALT) and its structural components has been described in different healthy animal species and in animals with diseases of the respiratory tract. In contrast to normal mammals, BALT is absent in healthy human adult lungs, but has been found in the lungs of children. The histological characteristics of organized mucosa-associated lymphoid tissue (MALT), its subsets of immune cells and their in situ distribution in the lung of healthy subadult and adult cattle shows close similarities with BALT in humans and other animal species such as sheep, horses and pigs. This study clearly demonstrates that organized MALT also occurs in the tracheal mucosa of cattle. The absence of tracheal MALT and BALT in calves suggest that these structures are not constitutive. In the mucosa of bovine trachea, bronchi and bronchioli, MHC II+ and CD11c+ dendritic cells (DCs) are located in the epithelium and in the lamina propria mucosae. These DCs are already present in calves soon after birth. Examination of tangential epithelial sheets shows that in the bovine tracheal epithelium, like in man and rat, a dense network of MHC II+ and CD11c+ DCs exists and that their number is considerably higher than in conventional transverse sections. In the bovine tracheal and bronchial epithelium, MHC II+ DCs are extending their dendrites towards the lumen indicating that these DCs possibly are involved in sampling of luminal antigens. The presence of significantly higher numbers of MHC II+ DCs in the tracheal and bronchial/bronchiolar mucosa of older cattle in than in calves possibly results from local stimulation with exogenous antigens during postnatal life. Detection of DCs expressing the costimulatory molecules CD80 and CD86 in calves and cattle suggests maturation of DCs, which is most likely induced by stimulation with exogenous antigens.

  View details for DOI 10.1111/ahe.12959

  View details for Web of Science ID 001077319900001

  View details for PubMedID 37646363

• T cell immunity ameliorates COVID-19 disease severity and provides post-exposure prophylaxis after peptide-vaccination, in Syrian hamsters FRONTIERS IN IMMUNOLOGY Somogyi, E., Kremlitzka, M., Csiszovszki, Z., Molnar, L., Lorincz, O., Toth, J., de Waal, L., Pattijn, S., Reineking, W., Beineke, A., Toke, E. R. 2023; 14: 1111629

  Abstract

  The emergence of novel SARS-CoV-2 variants that resist neutralizing antibodies drew the attention to cellular immunity and calls for the development of alternative vaccination strategies to combat the pandemic. Here, we have assessed the kinetics of T cell responses and protective efficacy against severe COVID-19 in pre- and post-exposure settings, elicited by PolyPEPI-SCoV-2, a peptide based T cell vaccine.75 Syrian hamsters were immunized subcutaneously with PolyPEPI-SCoV-2 on D0 and D14. On D42, hamsters were intranasally challenged with 102 TCID50 of the virus. To analyze immunogenicity by IFN-γ ELISPOT and antibody secretion, lymphoid tissues were collected both before (D0, D14, D28, D42) and after challenge (D44, D46, D49). To measure vaccine efficacy, lung tissue, throat swabs and nasal turbinate samples were assessed for viral load and histopathological changes. Further, body weight was monitored on D0, D28, D42 and every day after challenge.The vaccine induced robust activation of T cells against all SARS-CoV-2 structural proteins that were rapidly boosted after virus challenge compared to control animals (~4-fold, p<0.05). A single dose of PolyPEPI-SCoV-2 administered one day after challenge also resulted in elevated T cell response (p<0.01). The vaccination did not induce virus-specific antibodies and viral load reduction. Still, peptide vaccination significantly reduced body weight loss (p<0.001), relative lung weight (p<0.05) and lung lesions (p<0.05), in both settings.Our study provides first proof of concept data on the contribution of T cell immunity on disease course and provide rationale for the use of T cell-based peptide vaccines against both novel SARS-CoV-2 variants and supports post-exposure prophylaxis as alternative vaccination strategy against COVID-19.

  View details for DOI 10.3389/fimmu.2023.1111629

  View details for Web of Science ID 000924893300001

  View details for PubMedID 36761759

  View details for PubMedCentralID PMC9902696

• Infections with highly pathogenic avian influenza A virus (HPAIV) H5N8 in harbor seals at the German North Sea coast, 2021 EMERGING MICROBES & INFECTIONS Postel, A., King, J., Kaiser, F. K., Kennedy, J., Lombardo, M., Reineking, W., de le Roi, M., Harder, T., Pohlmann, A., Gerlach, T., Rimmelzwaan, G., Rohner, S., Striewe, L. C., Gross, S., Schick, L. A., Klink, J. C., Kramer, K., Osterhaus, A. E., Beer, M., Baumgaertner, W., Siebert, U., Becher, P. 2022; 11 (1): 725-729

  Abstract

  In brain tissue of three harbor seals of the German North Sea coast, high virus loads of highly pathogenic avian influenza virus (HPAIV) H5N8 were detected. Identification of different virus variants indicates high exposure to HPAIV circulating in wild birds, but there is no evidence for H5 specific antibodies in healthy seals. Replication of avian viruses in seals may allow HPAIV to acquire mutations needed to adapt to mammalian hosts as shown by PB2 627K variants detected in these cases.

  View details for DOI 10.1080/22221751.2022.2043726

  View details for Web of Science ID 000762625100001

  View details for PubMedID 35172704

  View details for PubMedCentralID PMC8890524

• Sox9, Hopx, and survivin and tuft cell marker DCLK1 expression in normal canine intestine and in intestinal adenoma and adenocarcinoma VETERINARY PATHOLOGY Reineking, W., Schauerte, I. E., Junginger, J., Hewicker-Trautwein, M. 2022; 59 (3): 415-426

  Abstract

  Self-renewal of the intestinal epithelium originates from stem cells located at the crypt base. Upregulation of various stem cell markers in intestinal epithelial neoplasms indicates a potential role of stem cells in tumorigenesis. In this study, the immunoreactivity of potential intestinal stem cell markers (Sry box transcription factor 9 [Sox9], homeodomain-only protein [Hopx], survivin) and tuft cell marker doublecortin-like kinase 1 (DCLK1) in normal canine intestine and intestinal epithelial neoplasms was investigated. Formalin-fixed paraffin-embedded (FFPE) small and large intestine as well as intestinal neoplasms (55 colorectal adenomas [CRAs], 17 small intestinal adenocarcinomas [SICs], and 12 colorectal adenocarcinomas [CRCs]) were analyzed immunohistologically. Potential stem cell markers Sox9, Hopx, and survivin were detected in the crypts of normal canine small and large intestine. DCLK1+ tuft cells were present in decreasing numbers along the crypt-villus axis of the jejunum and rarely detectable in large intestine. In canine intestinal epithelial tumors, nuclear Sox9 immunoreactivity was detectable in 84.9% (CRA), 80% (CRC), and 77% of epithelial neoplastic cells (SIC). Hopx and survivin were expressed within cytoplasm and nuclei of neoplastic cells in benign and malignant tumors. DCLK1 showed a cytoplasmic reaction within neoplastic cells. The combined score of Hopx, DCLK1, and survivin varied among the examined cases. Overall, malignant tumors showed lower DCLK1 scores but higher Hopx scores in comparison with benign tumors. For survivin, no differences were detectable. In conclusion, stem cell markers Sox9, Hopx, and survivin were detectable at the crypt base and the immunoreactivity of Sox9, DCLK1, survivin, and Hopx was increased in canine intestinal adenomas and adenocarcinomas compared with normal mucosa.

  View details for DOI 10.1177/03009858221079666

  View details for Web of Science ID 000772532400001

  View details for PubMedID 35220825

• Polyadenine insertion disrupting the <i>G6PC1</i> gene in German Pinschers with glycogen storage disease type Ia (GSD1A) ANIMAL GENETICS Christen, M., Reineking, W., Beineke, A., Jagannathan, V., Baumgaertner, W., Leeb, T. 2021; 52 (6): 900-902

  View details for DOI 10.1111/age.13146

  View details for Web of Science ID 000703656500001

  View details for PubMedID 34610166

  View details for PubMedCentralID PMC9293233

• Hanoverian F/W-line contributes to segregation of Warmblood fragile foal syndrome type 1 variant <i>PLOD1:c</i>.2032G&gt;A in Warmblood horses EQUINE VETERINARY JOURNAL Metzger, J., Kreft, O., Sieme, H., Martinsson, G., Reineking, W., Hewicker-Trautwein, M., Distl, O. 2021; 53 (1): 51-59

  Abstract

  Warmblood fragile foal syndrome (WFFS) is a lethal condition detected in Warmblood horses. Its origin and association with performance traits and fertility among horse populations is unknown.To validate the previously identified WFFS type 1 (WFFST1)-associated missense variant PLOD1:c.2032G>A and to investigate its distribution among various horses with particular focus on Hanoverian breed, as well as its pathomorphological picture. The study aimed at identifying the origin of the mutant allele and its correlation with performance and fertility traits in Warmblood horses.Retrospective case-control and association study.WFFST1 variant was validated using whole genome sequencing (WGS) in 78 equids. In an affected foal with a homozygous mutant genotype, necropsy was performed. Skin samples were examined using histology and transmission electron microscopy. Pathway analysis was performed to trace back 81 genetic carriers to the most common recent ancestor. Furthermore, generalised linear model analysis was employed to test estimated breeding values (EBVs) for differences in performance and fertility traits among different genotypes in Hanoverian horses.WFFST1 variant had the lowest minor allele frequency among all variants detected in WGS data in the region of PLOD1. Further genotyping of this variant revealed allele frequencies of 0.14 in Hanoverian horses. Histological investigations of the WFFST1-affected foal showed loosely arranged collagen fibres in the dermis. Ultrastructurally, multifocal areas with degraded collagen fibrils and fibrillar plaques were detected. Further pathway analysis revealed a stallion from the Hanoverian sire F/W line as the most common recent ancestor of all tested genetic carriers. Furthermore, WFFST1 variant was found to be correlated with EBVs for gait-related traits as well as conformation and dressage.Study evaluated carriers and cases only from Europe.This study provides a comprehensive evaluation of WFFST1 variant and traces it back to its potential origin.

  View details for DOI 10.1111/evj.13271

  View details for Web of Science ID 000533598600001

  View details for PubMedID 32323341

• Laryngeal chondritis as a differential for upper airway diseases in German sheep ACTA VETERINARIA SCANDINAVICA Reineking, W., Punsmann, T., Wagener, M., Verspohl, J., Ganter, M., Baumgaertner, W., Puff, C. 2020; 62 (1): 12

  Abstract

  Ovine laryngeal chondritis is a rare entity of sheep in the USA, Great Britain, New Zealand and Iceland, but has not been reported in Germany so far. Here, two German cases are reported.Two rams showed severe and progressive signs of dyspnea. Endoscopically, a severe bilateral swelling of the larynx was identified in both rams. Due to poor prognosis and progression of clinical signs one ram was euthanized, while the other ram died overnight. In both cases, a necrosuppurative laryngitis and chondritis of arytenoid cartilages was found at necropsy. Fusobacterium necrophorum and Streptococcus ovis were isolated from the laryngeal lesion in one animal.This is the first report of ovine laryngeal chondritis in continental Europe. This entity should be considered a differential diagnosis for upper airway disease in sheep.

  View details for DOI 10.1186/s13028-020-0510-0

  View details for Web of Science ID 000519913700001

  View details for PubMedID 32131871

  View details for PubMedCentralID PMC7057637

• Characterization of abortion, stillbirth and non-viable foals homozygous for the <i>Warmblood Fragile Foal Syndrome</i> ANIMAL REPRODUCTION SCIENCE Aurich, C., Mueller-Herbst, S., Reineking, W., Mueller, E., Wohlsein, P., Gunreben, B., Aurich, J. 2019; 211: 106202

  Abstract

  Warmblood fragile foal syndrome (WFFS) is a monogenetic defect with autosomal recessive inheritance. The WFFS homozygosity is non-compatible with extra-uterine life. Although as many as 15% of Warmblood horses are WFFS carriers, there has been little veterinary focus on this condition. The aim of this study was to determine outcomes and symptoms of clinical signs and pathological abnormalities during pregnancies when there were WFFS homozygous foetuses. Diagnostic material of 15 abortion or stillbirth cases with suspected diagnosis of WFFS was available for this study. Additionally, there were examinations in 37 cases where there were no indications of WFFS when submitted for routine diagnostic procedures. Foals in all cases were genotyped and external morphological defects were recorded. Amongst the 15 cases in which WFSS was suspected, there were 14 homozygous foetuses with the WFFS allele (WFFS/WFFS). Three heterozygous WFFS foetuses (N/WFFS) were detected in the cases submitted for routine diagnostic procedures. Of the 14 WFFS homozygous foetuses, 11 of mares had a gestation length of at least 320 days. Nine foals were born alive but died within a short time. Skin defects were obvious in 12 WFFS homozygous foals, and there was abnormal flexibility in the digital joints, flexed forelegs and incomplete closure of the abdominal wall in five, four, and one of the foals, respectively. In conclusion, the predominant manifestation of WFFS are death during the latter stages of gestation or live births with foals being non-viable. Losses in Warmblood horse breeding caused by WFFS are greater than previously assumed.

  View details for DOI 10.1016/j.anireprosci.2019.106202

  View details for Web of Science ID 000520009800002

  View details for PubMedID 31785623

• Predominance of Granular Cell Tumours among Testicular Tumours of Rabbits (<i>Oryctolagus cuniculi</i> f. dom.) JOURNAL OF COMPARATIVE PATHOLOGY Reineking, W., Seehusen, F., Lehmbecker, A., Wohlsein, P. 2019; 173: 24-29

  Abstract

  Testicular neoplasms are reported rarely in pet and laboratory rabbits (Oryctolagus cuniculi f. dom.), with interstitial cell tumours being the most commonly described testicular neoplasm. In this retrospective study, paraffin wax-embedded testicles with neoplastic changes from 52 rabbits were investigated. Five out of 52 animals exhibited more than one tumour type, resulting in a total of 57 tumours. Granular cell tumours were the most prevalent neoplasm with 36 examples (63%) out of the 57 testicular tumours. Interstitial cell tumours, Sertoli cell tumours and seminomas occurred less frequently. Granular cell tumours of the testis are rare in rabbits. Histological similarities between granular cell and interstitial cell (Leydig cell) tumours in haematoxylin and eosin-stained tissue sections may lead to misdiagnoses. The periodic acid-Schiff reaction or immunohistochemistry for periaxin and S100 protein, as well as ultrastructural analysis, are useful methods to confirm the diagnosis.

  View details for DOI 10.1016/j.jcpa.2019.09.012

  View details for Web of Science ID 000500939300005

  View details for PubMedID 31812170

• Ocular dermoids and microphthalmia in a juvenile TIERAERZTLICHE PRAXIS AUSGABE GROSSTIERE NUTZTIERE Meilwes, J., Leitzen, E., Reineking, W., Hewicker-Trautwein, M., Ganter, M. 2018; 46 (6): 379-384

  Abstract

  A 6-month-old Leine sheep was presented because of dermal tissue located on the left eye. During the first examination, the animal was clinically silent, apart from the deformed eye. A corneal and conjunctival dermoid and blindness of the left eye were diagnosed. Over a period of a year, the animal displayed conjunctivitis and inflammation of the affected eye. Furthermore, the sheep did not develop according to its age. During histopathological examination of the euthanized animal, microphthalmia and aphakia of the left eye were found in addition to the dermoids. Dermoids are described in humans and in different domestic animals. They can be combined with other congenital malformations. In sheep, dermoids are rarely diagnosed or reported in the literature.

  View details for DOI 10.15653/TPG-170690

  View details for Web of Science ID 000455804800005

  View details for PubMedID 30616279

• Canine primary jejunal and colonic epithelial cells predominantly express TLR5 and TLR9 but do not change TLR expression pattern after stimulation with certain Toll-like receptor ligands VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY Reineking, W., Junginger, J., Mischke, R., Hewicker-Trautwein, M. 2018; 206: 16-24

  Abstract

  The intestinal mucosa is in contact with abundant luminal antigens and coordinates immune responses to differentiate commensals from pathogens. Intestinal epithelial cells (IECs) not only represent a physical barrier but also an immunologically important cell type that recognizes microbe-associated molecular patterns via Toll-like receptors (TLR). The importance of TLR expression has been elucidated for intestinal disorders in humans, mice and dogs. However, as knowledge about canine intestinal TLRs is mainly limited to the transcriptional level, the present study analyzed the protein expression of TLR2, TLR3, TLR4, TLR5 and TLR9 by primary canine IECs in the steady state and after stimulation with TLR ligands. This exhibited TLR5 and TLR9 to be predominantly expressed in canine IECs. TLR stimulation did not result in changes of the TLR expression pattern. Further studies are needed to elucidate whether this implicates hyporesponsiveness of canine IECs towards TLR stimulation under steady state conditions.

  View details for DOI 10.1016/j.vetimm.2018.11.003

  View details for Web of Science ID 000453623100003

  View details for PubMedID 30502908