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.     

Sin Herbarium zu verfaufen

Ohne Zusammenfassung     

Safety and immunogenicity of a delta inulin-adjuvanted inactivated Japanese encephalitis virus vaccine in pregnant mares and foals

In 2011, following severe flooding in Eastern Australia, an unprecedented epidemic of equine encephalitis occurred in South-Eastern Australia, caused by Murray Valley encephalitis virus (MVEV) and a new variant strain of Kunjin virus, a subtype of West Nile virus (WNV_KUN). This prompted us to assess whether a delta inulin-adjuvanted, inactivated cell culture-derived Japanese encephalitis virus (JEV) vaccine (JE-ADVAX™) could be used in horses, including pregnant mares and foals, to not only induce immunity to JEV, but also elicit cross-protective antibodies against MVEV and WNV_KUN. Foals, 74–152 days old, received two injections of JE-ADVAX™. The vaccine was safe and well-tolerated and induced a strong JEV-neutralizing antibody response in all foals. MVEV and WNV_KUN antibody cross-reactivity was seen in 33% and 42% of the immunized foals, respectively. JE-ADVAX™ was also safe and well-tolerated in pregnant mares and induced high JEV-neutralizing titers. The neutralizing activity was passively transferred to their foals via colostrum. Foals that acquired passive immunity to JEV via maternal antibodies then were immunized with JE-ADVAX™ at 36–83 days of age, showed evidence of maternal antibody interference with low peak antibody titers post-immunization when compared to immunized foals of JEV-naïve dams. Nevertheless, when given a single JE-ADVAX™ booster immunization as yearlings, these animals developed a rapid and robust JEV-neutralizing antibody response, indicating that they were successfully primed to JEV when immunized as foals, despite the presence of maternal antibodies. Overall, JE-ADVAX™ appears safe and well-tolerated in pregnant mares and young foals and induces protective levels of JEV neutralizing antibodies with partial cross-neutralization of MVEV and WNV_KUN.     

Expression of Perilipin 2 (PLIN2) in Porcine Oocytes During Maturation

Perilipins have been reported to limit the interaction of lipases with neutral lipids within the droplets, thereby regulating neutral lipid accumulation and utilization. This study aimed to identify the location and expression of PLIN1 and PLIN2 in porcine oocytes during maturation. Quantitative real‐time polymerase chain reaction (qRT‐PCR), immunostaining and Western blot methods were used to characterize the expression and distribution patterns of PLIN1 and PLIN2 in porcine oocytes. The results showed that PLIN1 was not detectable in porcine oocytes. PLIN2 and BODIPY 493/503‐detected neutral lipid droplets appeared identical distribution patterns and extensive colocalization in both GV and MII porcine oocytes. PLIN2 protein expression was higher in GV oocytes than that in MII oocytes (p < 0.05), although PLIN2 mRNA expression was similar in both groups. These findings suggested that PLIN2 was a major lipid droplet‐associated protein in porcine oocytes.     

Dirofilaria immitis in cats from inner Sydney

Two hundred feral cats from the inner suburbs of Sydney were examined post mortem for adult Dirofilaria immitis and circulating microfilariae, and 101 of these cats were tested for heartworm antigens by an ELISA. Only 2 cats (1%) had adult heartworms, the blood sample from another cat contained a single microfilaria. The blood of a further three cats contained small amounts of D immitis antigen. Although D immitis occurs in cats in Sydney, the prevalence is not high enough to warrant prophylactic treatment.     

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.