In this study, we investigated whether β-glucan from Saccharomyces cerevisiae exerts beneficial effects on mucosal immunity in an ovine ruminal explant (ORE) model. Once the ORE model was established, viability was assessed through histological change, E-cadherin expression, CK-18 and Ki-67 distribution. Then, the OREs were co-cultured with β-glucan, following which, gene and protein expression levels of sheep β-defensin-1 (SBD-1), pro-inflammatory interleukin (IL)-6, and anti-inflammatory IL-10 were detected using quantitative real-time polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA). Hematoxylin & eosin staining, qPCR, and immunohistochemistry showed that the overall ORE structure was intact after 96 hours in culture, but explants cultured for more than 24 hours showed epithelial degradation. Therefore, we performed the follow-up test within 24 hours. qPCR and ELISA revealed that the gene and protein expression levels of SBD-1, IL-6, and IL-10 in the OREs significantly increased (P < 0.05) after treatment with β-glucan compared with controls. This study identified the feasibility and optimal conditions of ORE culture and demonstrated that β-glucan activates SBD-1, IL-6, and IL-10 secretion in OREs to promote mucosal immunity.

Immunohistochemistry has been used extensively to evaluate protein expression in clinical and research settings. However, immunohistochemistry is not always successful in veterinary medicine due to the lack of reliable antibody options, poor tissue preservation, labor-intensive staining, and antigen-retrieval optimization processes. RNAscope in-situ hybridization (ISH) is a powerful technology that uses a specific sequence probe to identify targeted mRNA. In this study, we demonstrate RNAscope ISH in 4 common canine malignancies, which are traditionally diagnosed by histopathology and immunohistochemistry. Probes were designed for commonly targeted mRNA markers of neoplastic tumors; these included c-kit in mast cell tumor, microphthalmia-associated transcription factor in malignant melanoma, ionized calcium-binding adapter molecule(-1) in histiocytic sarcoma, and alkaline phosphatase in osteosarcoma. A strong staining signal was obtained by these 4 targets in each canine malignancy. These results support the use of RNAscope ISH for definitive diagnosis in canine malignancies.

Bovine respiratory disease complex is etiologically complex and usually involves co-infection by several agents, including bovine parainfluenza virus-3 (BPIV-3), bovine respiratory syncytial virus (BRSV), and bovine coronavirus (BCoV). Traditionally, vaccines have been tested in seronegative calves infected with a single in vitro-passaged agent, often with little disease, resulting in unvaccinated subjects. To overcome the potential problem of attenuation coincident with in vitro culture of the viruses, cocktails of field isolates of BPIV-3s and BCoVs were passaged in the lungs of neonatal colostrum-deprived calves. Lung lavage fluids were used as inocula, alone and in combination with in-vivo passaged BRSV, and aerosolized into a trailer containing conventionally reared 9-week-old weaned Holstein calves with decayed, but still measurable, maternal antibodies. Calves developed acute respiratory disease of variable severity. Upon necropsy, there were characteristic gross and histologic lesions in the respiratory tract, associated immunohistochemically with BPIV-3, BRSV, and BCoV. In-vivo passage of viruses is an alternative to in vitro culture to produce inocula to better study the pathogenesis of infection and more rigorously and relevantly assess vaccine efficacy.

Integrin alpha-v/beta3 (αvβ3) recognizes arginine-glycine-aspartic acid (RGD) sequences and has important functions in cell adhesion, signaling, and survival. However, the expression of integrin αvβ3 in the equine lungs and jejunum is not well understood. The objective of this study was to explore the hitherto unknown expression of integrin αvβ3 in the lungs and jejuna of the horse using light and electron immunocytochemistry. Immunohistochemistry showed integrin αvβ3 on the epithelium, the immune cells in Peyer's patches, the smooth muscle, and the endothelium of equine jejuna. In equine lungs, we recognized integrin αvβ3 on the endothelium of blood vessels, the alveolar septa, the bronchial lymph nodes, and the cartilages, although the expression of integrin αvβ3 was weak on the epithelium of bronchioles. In conclusion, these are the first data to show the expression of integrin αvβ3 in equine lungs and jejuna.

Objective: This study was carried out to find out the immunolocalization of Interleukin 6 (IL-6) and Interleukin 10 (IL-10) in the testicular tissue of testicular dysfunction rat treated with secretome from human umbilical stem cells. Materials and methods: Rats were induced with cisplatin for testicular dysfunction condition. After that, the rats were grouped into two categories and were treated with secretome at 0.2 and 0.5 ml/kg BW once every week for 4 weeks. One week later, after the secretome treatment, the rats were sacrificed for histological evaluation using the immunohistochemical method. The preparation slides were examined using a light microscope and were analyzed descriptively and quantitatively. Results: There were no IL-6 and IL-10 immunoreactivities seen in the testicular tissue after cispla¬tin induction. However, the immunoreactivities of IL-6 and IL-10 were detected after secretome treatment, with both dosages of 0.2 and 0.5 ml/kg BW. These immunoreactivities were detected in the spermatogonia, spermatid/luminal tissue of seminiferous tubule, spermatogenic cells, and Leydig cells. In the cell calculation, the numbers of IL-6 immunoreactive cells were higher at the lower secretome dosage. In contrast, the numbers of IL-10 immunoreactive cells were higher at the higher secretome dosage. Conclusion: IL-6 and IL-10 can be localized in the testicular tissue of testicular dysfunction rat after secretome treatment. [J Adv Vet Anim Res 2020; 7(3.000): 514-520]

Objectives: Aqueous extracts of Rhizophora mucronata and Avicennia marina leaves were inves¬tigated for their hepatoprotective potential in diabetic rats. Materials and methods: One hundred twenty male albino rats were randomly assigned to eight equal groups (n = 15). The first group (control) comprised normal healthy rats, while the second to fifth groups were intraperitoneally injected with a single dose of streptozotocin (STZ) [60 mg/ kg body weight (BW)] for induction of diabetes. Group 2 was kept as positive diabetic control, while groups 35 were orally treated with aqueous extracts of R. mucronata (400 mg/kg BW), A. marina (400 mg/kg BW) and with a combination of ½ a dose of the two plants, respectively, for six weeks. Groups 68 were non-diabetic rats that orally received aqueous extracts of R. mucronata (400 mg/kg BW), A. marina (400 mg/kg BW), and a combination of ½ a dose of the two plants, respectively, for 6 weeks. Results: STZ-induced diabetic rats showed a significant reduction in serum glucose and liver enzymes, increased serum insulin, Homeostasis Model Assessment of &#946;-cells (HOMA-&#946;), and Homeostasis Model Assessment of Insulin Resistance (HOMA-IR). Histopathological and immuno¬histochemical examinations of the liver revealed improved pathologic criteria in the plant extract treated diabetic rats compared with the remarkable changes which had been seen in STZ-induced diabetic rats. Conclusion: This study suggests that the aqueous extract of R. mucronata or its combination with A. marina showed potent hypoglycemic and hepatoprotective effects for liver dysfunction, as well as histopathological and immunohistochemical changes in the liver of STZ-induced diabetic rats. [J Adv Vet Anim Res 2020; 7(1.000): 177-185]