Protective immunity enhanced Salmonella vaccine vectors delivering Helicobacter pylori antigens reduce H. pylori stomach colonization in mice.

Helicobacter pylori is a major cause of gastric mucosal inflammation, peptic ulcers, and gastric cancer. Emerging antimicrobial-resistant H. pylori has hampered the effective eradication of frequent chronic infections. Moreover, a safe vaccine is highly demanded due to the absence of effective vaccines against H. pylori. In this study, we employed a new innovative Protective Immunity Enhanced Salmonella Vaccine (PIESV) vector strain to deliver and express multiple H. pylori antigen genes. Immunization of mice with our vaccine delivering the HpaA, Hp-NAP, UreA and UreB antigens, provided sterile protection against H. pylori SS1 infection in 7 out of 10 tested mice. In comparison to the control groups that had received PBS or a PIESV carrying an empty vector, immunized mice exhibited specific and significant cellular recall responses and antigen-specific serum IgG1, IgG2c, total IgG and gastric IgA antibody titers. In conclusion, an improved S. Typhimurium-based live vaccine delivering four antigens shows promise as a safe and effective vaccine against H. pylori infection.

Therapeutic effects of equine amniotic membrane suspension on corneal re-epithelialization and haze in a modified lagomorph ex vivo wound healing model.

OBJECTIVE: To investigate the therapeutic effects of topical equine amniotic membrane (eAM) suspension following corneal wounding in a controlled experimental setting. PROCEDURES: Equine amniotic membrane was collected, gamma irradiated, homogenized for topical suspension preparation, and cryopreserved. Corneoscleral rims harvested from fresh rabbit globes were wounded via keratectomy and were maintained in an air-liquid interface ex vivo corneal culture model. Treatment groups included topical gamma irradiated eAM suspension (n = 20) and a control group (n = 20). Re-epithelialization of the wound was assessed with daily photographic evaluation of area of fluorescein uptake (mm2 ). Corneal wound haze after a 21-day period was assessed by photographic analysis of haze area (mm2 ) and pixel intensity (0-255). Histologic processing of corneal tissue was performed, and protein identification of eAM suspension using Liquid chromatography-mass spectrometry (LC-MS). RESULTS: The average day of complete corneal re-epithelialization in controls (5.5 ± 1.1) and topically treated (5.5 ± 0.6) corneas, and rates of reduction in area of fluorescein uptake over time did not significantly differ (p = .44). The corneal wound haze was significantly reduced in mean area by approximately 52% and intensity by 57% in corneas treated with topical eAM suspension (p < .05), compared to controls 21 days following wounding. Protein analysis identified numerous proteins, specifically decorin, dermatopontin, and lumican, which have previously been documented in eAM. CONCLUSIONS: Area and intensity of corneal wound haze were significantly reduced in corneas treated with gamma irradiated eAM suspension, which may be due to previously identified therapeutic proteins which promote corneal clarity.

Stimulation of regulatory T cells with Lactococcus lactis expressing enterotoxigenic E. coli colonization factor antigen 1 retains salivary flow in a genetic model of Sjögren's syndrome.

BACKGROUND: Sjögren's syndrome (SjS), one of the most common autoimmune diseases, impacts millions of people annually. SjS results from autoimmune attack on exocrine (salivary and lacrimal) glands, and women are nine times more likely to be affected than men. To date, no vaccine or therapeutic exists to treat SjS, and patients must rely on lifelong therapies to alleviate symptoms. METHODS: Oral treatment with the adhesin from enterotoxigenic Escherichia coli colonization factor antigen I (CFA/I) fimbriae protects against several autoimmune diseases in an antigen (Ag)-independent manner. Lactococcus lactis, which was recently adapted to express CFA/I fimbriae (LL-CFA/I), effectively suppresses inflammation by the induction of infectious tolerance via Ag-specific regulatory T cells (Tregs), that produce IL-10 and TGF-ß. To test the hypothesis that CFA/I fimbriae can offset the development of inflammatory T cells via Treg induction, oral treatments with LL-CFA/I were performed on the spontaneous, genetically defined model for SjS, C57BL/6.NOD-Aec1Aec2 mice to maintain salivary flow. RESULTS: Six-week (wk)-old C57BL/6.NOD-Aec1Aec2 mice were orally dosed with LL-CFA/I and treated every 3 wks; control groups were given L. lactis vector or PBS. LL-CFA/I-treated mice retained salivary flow up to 28 wks of age and showed significantly reduced incidence of inflammatory infiltration into the submandibular and lacrimal glands relative to PBS-treated mice. A significant increase in Foxp3+ and IL-10- and TGF-ß-producing Tregs was observed. Moreover, LL-CFA/I significantly reduced the expression of proinflammatory cytokines, IL-6, IL-17, GM-CSF, and IFN-γ. Adoptive transfer of CD4+ T cells from LL-CFA/I-treated, not LL vector-treated mice, restored salivary flow in diseased SjS mice. CONCLUSION: These data demonstrate that oral LL-CFA/I reduce or halts SjS progression, and these studies will provide the basis for future testing in SjS patients.

Retrospective analysis of ocular disease in a population of captive pteropodid bats, 2003-2020.

OBJECTIVE: To perform retrospective analysis of captive pteropodid bats presented to the University of Florida for ocular or adnexal disease from 2003-2020. ANIMALS STUDIED: Twenty-four individuals from seven species were included. PROCEDURES: Records were analyzed for disease process, methods of treatment, and surgical techniques and complications. RESULTS: The most frequently reported abnormality was corneal disease (79%), followed by cataracts (54%), and uveitis (42%). Corneal disease was primarily attributed to either trauma or exposure keratitis secondary to buphthalmia. The majority of uveitis appeared to be lens-induced. Five cases (21%) of glaucoma were reported, all of which accompanied lens luxation. Of the seven enucleations performed, six had post-operative complications (85.7%), including swelling at the surgical site, seroma formation, and bacterial infection. There was no significant relationship between age and trauma, age and cataract formation, sex and trauma, or species and cataract formation. CONCLUSIONS: The most common underlying cause of ocular pathology in these cases was trauma. While the bats tolerated topical and systemic treatment well, individual temperament must be taken into account when developing treatment plans, and prevention of injury is the most effective management strategy.

Retrobulbar embryonal tumor with multilayered rosettes in a golden retriever dog.

Although rare, embryonal tumors (previously called primitive neuroectodermal tumors) should be considered in the differential diagnosis of retrobulbar tumors in dogs regardless of the age of the patient, and ancillary tests are required for definitive diagnosis.

Pulmonary Sarcomatoid Carcinoma Associated with Arterial Thromboembolism in a Cat.

A 14-year-old, neutered male domestic shorthair cat presented for acute monoparesis with physical exam findings and biochemical data supportive of a distal arterial thromboembolism. Thoracic radiographs revealed an alveolar pattern in the right middle lung lobe and multifocal nodules in other lung lobes. A pulmonary mass was found on necropsy, which was composed of both carcinomatous and sarcomatous components, confirmed with cytokeratin and vimentin immunohistochemistry. Using the World Health Organization classification scheme for mixed pulmonary tumors, this tumor would be characterized as a pleomorphic squamous cell carcinoma under the umbrella term of pulmonary sarcomatoid carcinoma. The World Health Organization classification of mixed pulmonary tumors and its application to previously reported mixed pulmonary tumors in companion animals is discussed. This is the first reported case of this tumor type in a cat, as well as the first report of this tumor type associated with an arterial thromboembolism in any veterinary species.

Laparoscopic gonadectomy in a dog with 78,XX/78,XY chimerism and underdeveloped reproductive organs.

CASE DESCRIPTION: A 1-year-old externally sexually intact female Great Dane was referred for further evaluation of abnormal and underdeveloped internal reproductive organs. CLINICAL FINDINGS: Physical examination findings included a cranioventrally displaced vulva and a grade 2/6 left apical systolic heart murmur. No uterus or ovaries were identified during abdominal ultrasonography. Computed tomography with retrograde vaginourethrography revealed an underdeveloped uterus and possible left intra-abdominal gonad. Karyotyping revealed mixed sex chromosomes (70% XY and 30% XX). Analysis of a serum sample yielded positive results for anti-Müllerian hormone; other findings included mid range estradiol concentration (48.2 pg/mL [within reference intervals for sexually intact and neutered males and females]), low progesterone concentration (< 0.2 ng/mL [within reference intervals for anestrous females]), and low testosterone concentration (< 20 ng/dL [similar to the expected concentration in neutered males]). Overall, the results of the sex hormone analyses were consistent with findings for either a sexually intact female or a neutered male dog. The dog's cardiac structure and function were echocardiographically normal. TREATMENT AND OUTCOME: The dog was anesthetized and underwent laparoscopic gonadectomy. The gonads, although abnormal and underdeveloped, were readily identified intraoperatively and successfully removed. On the basis of histologic findings, the removed gonads were confirmed to be rudimentary testicles. The dog recovered from anesthesia and surgery without complications. CLINICAL RELEVANCE: Laparoscopic surgery was effective for visualization of abnormal and hypoplastic reproductive organs when abdominal ultrasonography and CT were of limited diagnostic usefulness, and laparoscopic surgery allowed straightforward gonadectomy in a 78,XX/78,XY chimeric dog.

Oral therapy with colonization factor antigen I prevents development of type 1 diabetes in Non-obese Diabetic mice.

Antigen (Ag)-specific tolerization prevents type 1 diabetes (T1D) in non-obese diabetic (NOD) mice but proved less effective in humans. Several auto-Ags are fundamental to disease development, suggesting T1D etiology is heterogeneous and may limit the effectiveness of Ag-specific therapies to distinct disease endotypes. Colonization factor antigen I (CFA/I) fimbriae from Escherichia coli can inhibit autoimmune diseases in murine models by inducing bystander tolerance. To test if Ag-independent stimulation of regulatory T cells (Tregs) can prevent T1D onset, groups of NOD mice were orally treated with Lactococcus lactis (LL) expressing CFA/I. LL-CFA/I treatment beginning at 6 weeks of age reduced disease incidence by 50% (p < 0.05) and increased splenic Tregs producing both IL-10 and IFN-γ 8-fold (p < 0.005) compared to LL-vehicle treated controls. To further describe the role of these Tregs in preventing T1D, protective phenotypes were examined at different time-points. LL-CFA/I treatment suppressed splenic TNF-α+CD8+ T cells 6-fold at 11 weeks (p < 0.005) and promoted a distinct microbiome. At 17 weeks, IFN-γ+CD4+ T cells were suppressed 10-fold (p < 0.005), and at 30 weeks, pancreatic Tbet+CD4+ T cells were suppressed (p < 0.05). These results show oral delivery of modified commensal organisms, such as LL-CFA/I, may be harnessed to restrict Th1 cell-mediated immunity and protect against T1D.

Regulating colonic dendritic cells by commensal glycosylated large surface layer protein A to sustain gut homeostasis against pathogenic inflammation.

Microbial interaction with the host through sensing receptors, including SIGNR1, sustains intestinal homeostasis against pathogenic inflammation. The newly discovered commensal Propionibacterium strain, P. UF1, regulates the intestinal immunity against pathogen challenge. However, the molecular events driving intestinal phagocytic cell response, including colonic dendritic cells (DCs), by this bacterium are still elusive. Here, we demonstrate that the glycosylation of bacterial large surface layer protein A (LspA) by protein O-mannosyltransferase 1 (Pmt1) regulates the interaction with SIGNR1, resulting in the control of DC transcriptomic and metabolomic machineries. Programmed DCs promote protective T cell response to intestinal Listeria infection and resist chemically induced colitis in mice. Thus, our findings may highlight a novel molecular mechanism by which commensal surface glycosylation interacting with SIGNR1 directs the intestinal homeostasis to potentially protect the host against proinflammatory signals inducing colonic tissue damage.

Primary leptomeningeal gliomatosis in a domestic shorthaired cat.

A 15-y-old neutered male domestic shorthaired cat was presented with a 16-d history of hindlimb paralysis in conjunction with 1-wk duration of inappetence and lethargy. Given intractable clinical signs, development of seizures, and poor prognosis, euthanasia was elected. Gross examination revealed mild, chronic, multifocal intervertebral disk disease; however, no gross abnormalities were noted in the spinal cord. Histologic examination of the cervical, thoracic, and lumbar spinal cord and the myelencephalon revealed diffuse and variable expansion of the meninges by sheets of neoplastic round-to-polygonal cells. The cells formed sheets and clusters, supported by a variably eosinophilic, fibrillar-to-basophilic, homogeneous matrix, and contained a small amount of eosinophilic cytoplasm. The nuclei were round with finely stippled to hyperchromatic chromatin and 1-2 small nucleoli. Mild white matter degeneration was present in the dorsal and ventral funiculi multifocally throughout the spinal cord, but was most severe in the ventral lumbar sections. Immunohistochemistry revealed strong intranuclear immunoreactivity for Olig2, and intracytoplasmic immunoreactivity for glial fibrillary acidic protein, MAP2, and vimentin in the neoplastic glial cells. To our knowledge, primary leptomeningeal gliomatosis has not been reported previously in a cat.

Anatomic characterization of pulmonary accessory fissures in canine cadavers.

Accessory fissures in the lungs are well described in humans, considered a normal finding, being identified in 60% of autopsied lungs, and more prevalent in the right lung (Gesase, ; Nene, Gajendra, & Sarma, 2011). In dogs, interlobar fissures are well recognized, but there is the lack of anatomic characterization for accessory fissures in the accessible literature. The purpose of this descriptive study was to identify the prevalence and to describe the anatomic localization of accessory fissures. The lungs from 87 dog cadavers used to teach veterinary gross anatomy were collected. Accessory fissures were characterized for each lung lobe, specific lung lobe location, orientation, length and the number of accessory fissures per lobe. Accessory fissures were recognized in 48/87 (55%) of canine lungs, all located in the periphery of the individual lobes. We found a significant association between the presence of accessory fissures and the costal surface of the lung (p < 0.0001), the right lung (p < 0.004), the right cranial lung lobe (p < 0.002) and the left cranial lung lobe (p < 0.04). Histologic results showed normal alveolar and respiratory bronchioles to the level of the accessory fissures. Our results show that accessory fissures are a common finding and should be considered a normal variant. They are more prevalent in the right lung, in the costal surface, and in the right and left cranial lung lobes. An anatomic nomenclature for accessory fissures based on the current Nomina Anatomica Veterinaria and the human literature is proposed. Further studies include a comparison among anatomical accessory fissures, and radiographic and computed tomographic images.

GRANULOMATOUS ENCEPHALOMYELITIS IN A FALSE GHARIAL (TOMISTOMA SCHLEGELII) ASSOCIATED WITH A NOVEL CHLAMYDIA SPECIES.

A 5-yr-old, captive, hatched, female false gharial (Tomistoma schlegelii) presented with a 1-mo history of cervical spinal curvature. Antemortem diagnostics, including blood work, electromyography, muscle biopsies, and advanced imaging tests, were either within reference ranges or did not identify any specific etiology. Necropsy revealed extensive, marked, chronic granulomatous encephalomyelitis along with neuronal necrosis, rarefaction, gliosis, and astrocytosis of the white and gray matter of the cerebrum, cerebellum, brainstem, and spinal cord. Pan-chlamydiae polymerase chain reaction protocols for the 16S ribosomal RNA and ompA genes were performed on samples of spinal cord and brain, and both resulted in amplicons. Sequencing of the products revealed that they were positive for a novel Chlamydia species. Infections by members of the phylum Chlamydiae have been reported in a diverse range of vertebrate hosts, including crocodilians. Chlamydia spp. infections are likely underdiagnosed because of a paucity of diagnostic techniques specific for detection. This is the first case report of a novel Chlamydia species associated with severe granulomatous encephalomyelitis in a false gharial.

Clinical behavior of intraocular teratoid medulloepithelioma in two-related Quarter Horses.

The objective of this paper is to describe clinical behavior, histopathologic features, and immunohistochemical staining of two-related horses with intraocular teratoid medulloepithelioma. Two-related Quarter Horses with similar intraocular masses presented to the UF-CVM Comparative Ophthalmology Service for evaluation and treatment. The first horse, a 3-year-old gelding, had glaucoma and a cyst-like mass in the anterior chamber. Enucleation was performed. Histopathology revealed a teratoid medulloepithelioma. The tumor was considered to be completely excised. Fifteen months later, the gelding presented with swelling of the enucleated orbit and local lymph nodes with deformation of the skull. Cytology revealed neuroectodermal neoplastic cells. Necropsy confirmed tumor metastasis. Six weeks later, a 9-year-old mare, a full sibling to the gelding, presented for examination. An infiltrative mass of the iris and ciliary body was found that extended into the anterior, posterior, and vitreal chambers. Uveitis was present, but secondary glaucoma was not noted. Enucleation was performed and the histopathologic diagnosis was also teratoid medulloepithelioma. The mare has had no recurrence to date, 2 years following enucleation. Metastasis of intraocular teratoid medulloepithelioma is possible. Staging is recommended in cases where the diagnosis of teratoid medulloepithelioma is confirmed. Surveillance of full siblings is recommended until more information regarding etiology is known.

Functional metastatic parathyroid adenocarcinoma in a dog.

A 12-year-old dachshund dog was presented for persistent hypercalcemia and hyperparathyroidism despite bilateral parathyroidectomy. Magnetic resonance imaging of the head, neck, and cranial mediastinum identified an increased number of cranial mediastinal lymph nodes with heterogeneous signal intensity. Hypercalcemia and hyperparathyroidism resolved after surgery to remove multiple cranial mediastinal lymph nodes, one of which contained presumed metastatic parathyroid tissue.

Adénocarcinome parathyroïdien métastatique fonctionnel chez un chien. Un chien Dachsund âgé de 12 ans a été présenté pour de l'hypercalcémie et de l'hyperparathyroïdie persistantes malgré une parathyroïdectomie bilatérale. Une imagerie par résonance magnétique de la tête, du cou et du médiastin crânien a identifié un nombre accru de ganglions lymphatiques médiastinaux avec une intensité hétérogène du signal. L'hypercalcémie et l'hyperparathyroïdie se sont résorbées après la chirurgie pour enlever les nombreux ganglions lymphatiques médiastinaux crâniens, dont l'un contenait du tissu parathyroïdien métastatique présumé.(Traduit par Isabelle Vallières).

Effects of aurothiomalate treatment on canine osteosarcoma in a murine xenograft model.

Osteosarcoma is a highly fatal cancer, with most patients ultimately succumbing to metastatic disease. The purpose of this study was to evaluate the effects of the antirheumatoid drug aurothiomalate on canine and human osteosarcoma cells and on canine osteosarcoma growth and metastasis in a mouse xenograft model. We hypothesized that aurothiomalate would decrease osteosarcoma cell survival, tumor cellular proliferation, tumor growth, and metastasis. After performing clonogenic assays, aurothiomalate or a placebo was administered to 54 mice inoculated with canine osteosarcoma. Survival, tumor growth, embolization, metastasis, histopathology, cell proliferation marker Ki67, and apoptosis marker caspase-3 were compared between groups. Statistical analysis was carried out using the Kaplan-Meier method with the log-rank test and one-way analysis of variance with the Tukey's test or Dunn's method. Aurothiomalate caused dose-dependent inhibition of osteosarcoma cell survival (P<0.001) and decreased tumor growth (P<0.001). Pulmonary macrometastasis and Ki67 labeling were reduced with low-dose aurothiomalate (P=0.033 and 0.005, respectively), and tumor emboli and pulmonary micrometastases were decreased with high-dose aurothiomalate (P=0.010 and 0.011, respectively). There was no difference in survival, tumor development, ulceration, mitotic indices, tumor necrosis, nonpulmonary metastases, and caspase-3 labeling. Aurothiomalate treatment inhibited osteosarcoma cell survival and reduced tumor cell proliferation, growth, embolization, and pulmonary metastasis. Given aurothiomalate's established utility in canine and human medicine, our results suggest that this compound may hold promise as an adjunctive therapy for osteosarcoma. Further translational research is warranted to better characterize the dose response of canine and human osteosarcoma to aurothiomalate.

Radiographic characterization of presumed plate-like atelectasis in 75 nonanesthetized dogs and 15 cats.

Discrete discoid or linear areas of increased soft opacity have been observed within the pulmonary parenchyma in thoracic radiographs of dogs and cats. Similar radiographic findings have been described in humans and termed plate-like atelectasis. The purpose of this retrospective study was to describe locations and characteristics of presumed plate-like atelectasis, presence of concurrent thoracic disease(s), and presence of persistent pulmonary changes on recheck thoracic radiographic studies in a cohort of dogs and cats. Hospital records between 2004 and 2011 were searched and a total of 90 cases were included (75 dogs and 15 cats, 2-17 years of age). Plate-like atelectasis was most commonly found in left lateral radiographs. Plate-like atelectasis was observed in the cranial thorax and was oriented in a dorsocranial to ventrocaudal direction in 68 (75%) patients. Plate-like atelectasis averaged 29.6 ± 14.4 mm in length and 2.6 ± 1.3 mm in width. In 57 of the 90 patients (63%), plate-like atelectasis was the only abnormality found. Plate-like atelectasis was present in 7 of 22 cases where follow-up radiographs were available. Findings from the current study indicated that, while the etiology of plate-like atelectasis remains unknown, anatomic variations in sublobar pulmonary anatomy might account for pleural areas of atelectasis. The authors propose that the presence of plate-like atelectasis may represent areas of atelectasis that track along sublobar lung lobe separations, an area of hypoventilation or decreased collateral ventilation, and/or area of decreased localized surfactant deficiency.