Actinomyces hyovaginalis-associated lymphadenitis in a Nubian goat.

A 6-year-old Nubian goat with a history of progressive weight loss and cough was presented for necropsy. The goat tested negative for antibodies to caseous lymphadenitis and caprine arthritis and encephalitis by hemagglutination inhibition assay and enzyme-linked immunosorbent assay, respectively. Postmortem examination revealed marked enlargement and, with histopathology, a fibrinopurulent necrotizing lymphadenitis of a tracheobronchial lymph node, with an appearance similar to that reported in cases of caseous lymphadenitis. An organism characterized by molecular methods as Actinomyces hyovaginalis was isolated together with Staphylococcus spp. and Streptococcus spp. from the lesion. No Corynebacterium pseudotuberculosis was recovered. To the authors' knowledge, this is the first isolation of A. hyovaginalis from a goat. Although the exact contribution of A. hyovaginalis to the lesion remains to be established, this case demonstrates that A. hyovaginalis should be considered in cases of caseous lymphadenitis-type lesions, especially when C. pseudotuberculosis has been excluded.

Culture conditions for equine bone marrow mesenchymal stem cells and expression of key transcription factors during their differentiation into osteoblasts.

BACKGROUND: The use of equine bone marrow mesenchymal stem cells (BMSC) is a novel method to improve fracture healing in horses. However, additional research is needed to identify optimal culture conditions and to determine the mechanisms involved in regulating BMSC differentiation into osteoblasts. The objectives of the experiments were to determine: 1) if autologous or commercial serum is better for proliferation and differentiation of equine BMSC into osteoblasts, and 2) the expression of key transcription factors during the differentiation of equine BMSC into osteoblasts. Equine BMSC were isolated from the sterna of 3 horses, treated with purchased fetal bovine serum (FBS) or autologous horse serum (HS), and cell proliferation determined. To induce osteoblast differentiation, cells were incubated with L-ascorbic acid-2-phosphate and glycerol-2-phosphate in the presence or absence of human bone morphogenetic protein2 (BMP2), dexamethasone (DEX), or combination of the two. Alkaline phosphatase (ALP) activity, a marker of osteoblast differentiation, was determined by ELISA. Total RNA was isolated from differentiating BMSC between d 0 to 18 to determine expression of runt-related transcription factor2 (Runx2), osterix (Osx), and T-box3 (Tbx3). Data were analyzed by ANOVA. RESULTS: Relative to control, FBS and HS increased cell number (133 ± 5 and 116 ± 5%, respectively; P < 0.001) and 5-bromo-2'-deoxyuridine (BrdU) incorporation (167 ± 6 and 120 ± 6%, respectively; P < 0.001). Treatment with DEX increased ALP activity compared with control (1,638 ± 38%; P < 0.001). In the absence and presence of Dex, BMP-2 did not alter ALP activity (P > 0.8). Runt-related transcription factor2 expression increased 3-fold (P < 0.001) by d 6 of culture. Osterix expression increased 9-fold (P < 0.05) by d 18 of culture. Expression of Tbx3 increased 1.8-fold at d 3 (P < 0.01); however expression was reduced 4-fold at d 18 (P < 0.01). CONCLUSIONS: Dexamethasone, but not BMP-2, is required for differentiation of equine BMSC into osteoblasts. In addition, expression of Runx2 and osterix increased and expression of Tbx3 is reduced during differentiation.