[Establishment of Simple and Quick Method to Confirm the Heating History of Milk and Dairy Products].

In Japan, the import quarantine procedure for dairy products was newly introduced in November 2017. The treatment such as 15 sec heating at 72℃ is required for virus inactivation when importing milk or dairy products from the area which is not free from foot and mouth disease. Moreover, the heating history of imported items is also inspected as import quality procedures. The IDF 63 method is known as one of the methods to confirm the heating history of milk by checking the alkaline phosphatase (ALP) activity. However, this procedure is complicated for daily quarantine inspection. Therefore, we attempted the ALP activity measurement based on the amount of fluorescent substance produced by the enzymatic reaction. Milk and dairy products derived from cow, sheep, and goat were tested after various heat treatment conditions. The ALP of heat-treated milk and dairy products derived from these species above were confirmed to be inactivated under substantially the same heat treatment for 15 sec at 72℃. The measurement method established in this study is simpler, faster, and requires smaller amount of sample compared to other methods. Additionally, the method was also applicable to confirm the heating history of various dairy products by making them into suspension.

Systemic Staphylococcus pseudintermedius infection in an arctic fox (Vulpes lagopus) with severe multifocal suppurative meningoencephalitis and nephritis.

A 2-year-female arctic fox (Vulpes lagopus) developed anorexia, dehydration, and emaciation during the quarantine period for importation from Norway, and died 17 days later. At necropsy, a fistula was observed on the left gluteal region, and the left eye, left brain, and kidneys were discolored. Histologically, severe diffuse suppurative meningoencephalitis and renal abscesses were detected. Numerous Gram-positive cocci were detected in these lesions. Multidrug-susceptible Staphylococcus pseudintermedius were isolated from the lesions. These results suggest that S. pseudintermedius can cause severe multifocal suppurative meningoencephalitis and nephritis in foxes. This is the first report of multidrug-susceptible S. pseudintermedius meningoencephalitis and nephritis in a fox.

Characterizing the temporal patterns of avian influenza virus introduction into Japan by migratory birds.

The objectives of the present study were to observe the temporal pattern of avian influenza virus (AIV) introduction into Japan and to determine which migratory birds play an important role in introducing AIV. In total, 19,407 fecal samples from migratory birds were collected at 52 sites between October 2008 and May 2015. Total nucleic acids extracted from the fecal samples were subjected to reverse transcription loop-mediated isothermal amplification to detect viral RNA. Species identification of host migratory birds was conducted by DNA barcoding for positive fecal samples. The total number of positive samples was 352 (prevalence, 1.8%). The highest prevalence was observed in autumn migration, and a decrease in prevalence was observed. During autumn migration, central to southern Japan showed a prevalence higher than the overall prevalence. Thus, the main AIV entry routes may involve crossing the Sea of Japan and entry through the Korean Peninsula. Species identification was successful in 221 of the 352 positive samples. Two major species sequences were identified: the Mallard/Eastern Spot-billed duck group (115 samples; 52.0%) and the Northern pintail (61 samples; 27.6%). To gain a better understanding of the ecology of AIV in Japan and the introduction pattern of highly pathogenic avian influenza viruses, information regarding AIV prevalence by species, the prevalence of hatch-year migratory birds, migration patterns and viral subtypes in fecal samples using egg inoculation and molecular-based methods in combination is required.