A filter-assisted culture method for isolation of Treponema spp. from bovine digital dermatitis and their identification by MALDI-TOF MS

Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. (T. pedis, T. phagedenis, and T. medium) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS–based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.     

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MicroRNA-21 expression,serum tumor markers,and immunohistochemistry in canine mammary tumors

Background

Canine mammary tumors (CMTs) are one of the most common malignancies in dogs and are associated with significant mortality. Serum tumor markers and non-coding microRNAs have gained widespread popularity in human oncology studies. The present study has two aims, first one is to investigate the miR-21 expression compared with changes in serum tumor markers (CEA and CA15-3) in CMT. The second aim is to detect the immunohistochemistry markers as vimentin, P63, and -SMA in CMT.

Methods

This study enrolled 17 female dogs: 10 with mammary tumors and seven controls without tumors. Blood samples were collected to measure miR-21, CEA, and CA 15-3, and histological samples were prepared for histological grading and immunohistochemistry.

Results

CA 15-3 was elevated in all animals, whereas CEA levels showed no change compared with controls. miR-21 was upregulated 12.84-fold in animals with CMT. The most frequently recorded CMT was the mixed type. Myoepithelial cells were identified by P63 immunoreactivity, but not SMA. High expression of miR-21 was observed with positive vimentin immunoreactivity, indicating the mesenchymal origin of the tumor cells.

Conclusion

The present study showed that miR-21 was elevated to a greater extent than CA 15-3 (12.84-fold vs. threefold). Tumors that was positive for vimentin immunoreactivity was also associated with an elevation in the levels of miR-21, showing that miR-21 is released from mesenchymal cells. These findings support the hypothesis that miR-21 may be a more sensitive, noninvasive indicator for CMT.

     

Effect of a single injection of gonadotropin-releasing hormone (GnRH) and human chorionic gonadotropin (hCG) on testicular blood flow measured by color doppler ultrasonography in male Shiba goats

Although color Doppler ultrasonography has been used to evaluate testicular blood flow in many species, very little has been done in goat. Eight male Shiba goats were exposed to a single intramuscular injection of either gonadotropin-releasing hormone (GnRH group; 1 µg/kg BW) or human chorionic gonadotropin (hCG group; 25 IU/kg BW). Plasma testosterone (T), estradiol (E2) and inhibin (INH) were measured just before (0 hr) and at different intervals post injection by radioimmunoassay. Testis volume (TV) and Doppler indices, such as resistive index (RI) and pulsatility index (PI) of the supratesticular artery, were measured by B-mode and color Doppler ultrasonography, respectively. The results indicated an increase in testicular blood flow in both groups, as RI and PI decreased significantly (P<0.05), but this increase was significant higher and earlier in hCG group (1 hr) than in the GnRH group (2 hr). A high correlation was found for RI and PI with both T (RI, r= −0.862; PI, r= −0.707) and INH in the GnRH group (RI, r=0.661; PI, r=0.701). However, a significant (P<0.05) correlation was found between E2 and both RI (r= −0.610) and PI (r= −0.763) in hCG group. In addition, TV significantly increased and was highly correlated with RI in both groups (GnRH, r= −0.718; hCG, r= −0.779). In conclusion, hCG and GnRH may improve testicular blood flow and TV in Shiba goats.     

Effect of different pre-sowing seed treatments on the germination of <Emphasis Type="Italic">Leucaena leucocephala</Emphasis> (Lam.) and <Emphasis Type="Italic">Acacia farnesiana</Emphasis> (L.)

Leucaena leucocephala and Acacia farnesiana are tree species used for several agricultural purposes in the Mediterranean region. The seeds of these species exhibit dormancy, causing delayed germination. Two experiments were conducted to investigate the effect of pre-sowing treatments (scarification, hot water, or soaking) on seed germination of L. leucocephala and A. farnesiana. In one experiment, seeds were exposed to three pre-sowing treatments: control, sandpaper scarification, or soaking in 70°C water for 4, 8, 12, 16, 20, or 24 min. In another experiment, seeds were soaked in 70°C water for 20 min, and then soaked in water at room temperature for an additional 24, 48, or 72 h or blade scarified. In general, soaking the seeds of the two species in hot water was more effective in breaking seed dormancy than scarification. Sandpaper scarification was not effective for either species. Blade scarification increased A. farnesiana seed germination to 56%, indicating that seed dormancy was mainly a consequence of hardseededness. L. leucocephala seeds collected from Jordan University of Science and Technology (JUST) site and soaked in 70°C water for 20 min and then soaked for 24, 48, or 72 h had germination rates above 97%. Our results suggest that blade scarification of A. farnesiana seeds and soaking of L. leucocephala seeds in 70°C water for 20 min are effective treatments to break seed dormancy and enhance seed germination of these vital species.