Effect of hydrocolloid-nitric oxide wound dressings on wound healing in dogs.

OBJECTIVE: To determine the rates of wound healing in surgically created wounds between nitric oxide releasing wound dressings and control wound dressings. STUDY DESIGN: Prospective, controlled, randomized experimental study. ANIMALS: Purpose-bred, adult, male Beagles (n = 6). METHODS: Four 2 × 2 cm wounds were surgically created on the trunk of each dog with each wound randomized to treatment with a nitric oxide wound pad (NP), nitric oxide wound gel (NG), plain hydrocolloid wound dressing (HC), or Telfa pad (T). Wound images were taken daily for 8 days then every other day until day 21 with images masked and randomized for evaluation. Total wound area, contraction percentage, and days until granulation were calculated. RESULTS: Time to first appearance of granulation tissue was significantly shorter for NP (3.2 days) than for NG (4 days; p = .023), HC (4.5 days; p = .001), and T (5.2 days; p < .0001). There were significant differences in total wound area and contraction percentage between sites and treatments (p < .001). Total wound area for NG was lower than treatment T (0.7 ± 0.1 cm3; p < .001), HC (0.9 ± 0.1 cm3, p < .001), and NP (0.6 ± 0.1 cm3, p < .001). CONCLUSION: Use of a nitric oxide wound dressing resulted in faster wound healing as evidenced by lower total wound area and higher contraction in the NG group and faster time to granulation tissue development in the NP group. CLINICAL SIGNIFICANCE: Nitric oxide wound dressings are innovative and inexpensive products that can significantly decrease the amount of time and cost necessary for open or second intention wound resolution in dogs.

Piscichuvirus-Associated Severe Meningoencephalomyelitis in Aquatic Turtles, United States, 2009-2021.

Viruses from a new species of piscichuvirus were strongly associated with severe lymphocytic meningoencephalomyelitis in several free-ranging aquatic turtles from 3 coastal US states during 2009-2021. Sequencing identified 2 variants (freshwater turtle neural virus 1 [FTuNV1] and sea turtle neural virus 1 [STuNV1]) of the new piscichuvirus species in 3 turtles of 3 species. In situ hybridization localized viral mRNA to the inflamed region of the central nervous system in all 3 sequenced isolates and in 2 of 3 additional nonsequenced isolates. All 3 sequenced isolates phylogenetically clustered with other vertebrate chuvirids within the genus Piscichuvirus. FTuNV1 and STuNV1 shared ≈92% pairwise amino acid identity of the large protein, which narrowly places them within the same novel species. The in situ association of the piscichuviruses in 5 of 6 turtles (representing 3 genera) with lymphocytic meningoencephalomyelitis suggests that piscichuviruses are a likely cause of lymphocytic meningoencephalomyelitis in freshwater and marine turtles.

Recognition of Field Signs, Necropsy Procedures, and Evaluation of Macroscopic Lesions of Cervids Infected with Epizootic Hemorrhagic Disease Virus.

Epizootic hemorrhagic disease virus (EHDV) is an arthropod-borne RNA virus in the genus Orbivirus, family Sedoreoviridae. Globally, seven known EHDV serotypes circulate among ruminant hosts and Culicoides species vectors. A variety of domestic and wild ruminant species are susceptible to EHDV infection, but infection outcome is highly variable between species, as well as between individuals of the same species. Thus, this disease system inherently operates at the wildlife-livestock interface. Domestic cattle are important hosts for EHDV, and while inapparent infection is the most common outcome, reports of clinical disease have increased in some parts of the world. However, fatal infection of cattle is rare. Among wildlife, white-tailed deer (Odocoileus virginianus) are highly susceptible to severe and often fatal disease. Considering the paucity of data and poorly characterized pathology of EHD in cattle, white-tailed deer represent a case study for describing the field signs and necropsy lesions associated with EHD. Here we describe the field signs that commonly define EHD outbreaks in North America, a basic approach to a gross necropsy examination of white-tailed deer, description of the gross lesions that may be present, and diagnostic sample collection. Field investigations of large-scale EHD outbreaks are common in North America. The necropsy examination is an essential tool in the study of disease and when coupled with other disciplines (e.g., virology, immunology, epidemiology) has been fundamentally important to understanding EHD in North America.

Microarray Gene Expression Analysis of Lesional Skin in Canine Pemphigus Foliaceus.

Canine pemphigus foliaceus (PF) is considered the most common autoimmune skin disease in dogs; the mechanism of PF disease development is currently poorly understood. Therefore, this study aimed to characterize the molecular mechanisms and altered biological pathways in the skin lesions of canine PF patients. Using an RNA microarray on formalin-fixed, paraffin-embedded samples, we analyzed the transcriptome of canine PF lesional skin (n = 7) compared to healthy skin (n = 5). Of the 800 genes analyzed, 420 differentially expressed genes (DEGs) (p < 0.05) were found. Of those, 338 genes were significantly upregulated, including pro-inflammatory and Th17-related genes. Cell type profiling found enhancement of several cell types, such as neutrophils, T-cells, and macrophages, in PF skin compared to healthy skin. Enrichment analyses of the upregulated DEGs resulted in 78 statistically significant process networks (FDR < 0.05), including the Janus kinase signal transducer and activator of transcription (JAK-STAT) and mitogen-activated protein kinase (MAPK) signaling. In conclusion, canine PF lesional immune signature resembles previously published changes in human pemphigus skin lesions. Further studies with canine PF lesional skin using next-generation sequencing (e.g., RNA sequencing, spatial transcriptomics, etc.) and the development of canine keratinocyte/skin explant PF models are needed to elucidate the pathogenesis of this debilitating disease.

Development of a peptide-generated antibody to rabbit hemorrhagic disease virus 2 VP60 and its immunohistochemical application in natural cases.

Rabbit hemorrhagic disease virus 2 (RHDV2) has spread across the United States infecting and causing death in domestic and wild rabbits. Immunohistochemistry (IHC) would be a useful tool for the detection of RHDV2 antigen in tissues as it is inexpensive and readily achievable in most diagnostic laboratories. However, there is no readily available antibody for this purpose. To fill this void, we generated an RHDV2 capsid protein VP60-specific antibody in chicken eggs and validated the antibody using formalin-fixed tissues from 5 domestic rabbits naturally infected with RHDV2. Viral antigen was detected immunohistochemically in various tissues, most prominently in hepatocytes and macrophages in liver, and in macrophages in spleen and cecal lymphoid tissue. Intravascular mononuclear cells in lung and renal tubular and biliary epithelium also were immunolabeled. Both nuclear and cytoplasmic immunolabeling were observed. This peptide-generated antibody is a potentially useful tool as an adjunct to reverse-transcription PCR or in situ hybridization for detection of RHDV2 in tissues.

Swine influenza A virus isolates containing the pandemic H1N1 origin matrix gene elicit greater disease in the murine model.

Since the 1990s, endemic North American swine influenza A viruses (swFLUAVs) contained an internal gene segment constellation, the triple reassortment internal gene (TRIG) cassette. In 2009, the H1N1 pandemic (pdmH1N1) virus spilled back into swine but did not become endemic. However, the pdmH1N1 contributed the matrix gene (pdmM) to the swFLUAVs circulating in the pig population, which replaced the classical swine matrix gene (swM) found in the TRIG cassette, suggesting the pdmM has a fitness benefit. Others have shown that swFLUAVs containing the pdmM have greater transmission efficiency compared to viruses containing the swM gene segment. We hypothesized that the matrix (M) gene could also affect disease and utilized two infection models, resistant BALB/c and susceptible DBA/2 mice, to assess pathogenicity. We infected BALB/c and DBA/2 mice with H1 and H3 swFLUAVs containing the swM or pdmM and measured lung virus titers, morbidity, mortality, and lung histopathology. H1 influenza strains containing the pdmM gene caused greater morbidity and mortality in resistant and susceptible murine strains, while H3 swFLUAVs caused no clinical disease. However, both H1 and H3 swFLUAVs containing the pdmM replicated to higher viral titers in the lungs and pdmM containing H1 viruses induced greater histological changes compared to swM H1 viruses. While the surface glycoproteins and other gene segments may contribute to swFLUAV pathogenicity in mice, these data suggest that the origin of the matrix gene also contributes to pathogenicity of swFLUAV in mice, although we must be cautious in translating these conclusions to their natural host, swine. IMPORTANCE: The 2009 pandemic H1N1 virus rapidly spilled back into North American swine, reassorting with the already genetically diverse swFLUAVs. Notably, the M gene segment quickly replaced the classical M gene segment, suggesting a fitness benefit. Here, using two murine models of infection, we demonstrate that swFLUAV isolates containing the pandemic H1N1 origin M gene caused increased disease compared to isolates containing the classical swine M gene. These results suggest that, in addition to other influenza virus gene segments, the swFLUAV M gene segment contributes to pathogenesis in mammals.

Pandemic Risk Assessment for Swine Influenza A Virus in Comparative In Vitro and In Vivo Models.

Swine influenza A viruses pose a public health concern as novel and circulating strains occasionally spill over into human hosts, with the potential to cause disease. Crucial to preempting these events is the use of a threat assessment framework for human populations. However, established guidelines do not specify which animal models or in vitro substrates should be used. We completed an assessment of a contemporary swine influenza isolate, A/swine/GA/A27480/2019 (H1N2), using animal models and human cell substrates. Infection studies in vivo revealed high replicative ability and a pathogenic phenotype in the swine host, with replication corresponding to a complementary study performed in swine primary respiratory epithelial cells. However, replication was limited in human primary cell substrates. This contrasted with our findings in the Calu-3 cell line, which demonstrated a replication profile on par with the 2009 pandemic H1N1 virus. These data suggest that the selection of models is important for meaningful risk assessment.