Activities are mainly divided into three, namely; lecture, laboratory practical and field trip. Aside from Ryukoku University, lectures and field visits were composed of diverse topics done at Yokohama Plant Protection, Institute of Plant Protection in NARO, Kyushu Okinawa Agricultural Research Center, NARO, Kumamoto University, Nara Institute of Science and Technology, Kyoto University, and Forestry and Fishery Product Research Institute.Laboratory activities include DNA extraction using ISOHAIR, PCR protocols following the Mi, Harris and Power methods, gel electrophoresis, restriction fragment length polymorphism (RFLP), brilliant blue staining for RKN-infected plants and picking of stained RKN eggs, extraction of cyst nematodes using the Fenwick funnel method, quantification of lesion nematodes (Pratylenchus spp.) using real-time PCR, and the use of Loop-mediated isothermal amplification (LAMP) for the detection of pine wilt nematode, Bursaphelenchus xylophilus.
Important findings regarding the attachment activities were presented during the activity evaluation report held at Ryukoku University on April 3, 2023. The evaluation report was attended by undergraduate and graduate students under the supervision of Prof. Iwahori. Both participants were able to discuss the usefulness or advantages, differences, and possible scenarios which resulted in inconsistency of expected results. In these activities which involved technical skills and execution of optimized protocols for morphological and molecular identification and detection of specific nematodes, participants were able to give conclusions and recommendations that are applicable to their laboratories and nature of work.Also, new information was generated such as the detection of Hirschmanniella spp. on lotus that must be monitored for possible cross-infestation to other crops particularly aquatic plants which have gained popularity in the international trade. Another is the potential application of LAMP to identify plant-parasitic nematodes particularly B. xylophilus which is a global threat to pine trees.
Recommendations coming from the participants were also discussed during the evaluation report. Priority on the list is the continuous and intense training on nematodes for ASEAN plant protectionists to gain skills, new techniques as well as updated pest lists and of global quarantine importance. Second, is the monitoring of staff and country performance in terms of their capacity in handling diagnostics on plant parasitic nematodes to further equip and assist them in establishing and sustaining the ASEAN Diagnostics Network.Third, it was suggested to provide equipment for the diagnostic laboratory. However, as per discussion, the need for infrastructure and/or equipment can be proposed to other funding agencies with the assistance from APHCN-ASEANET in terms of project proposal preparation, since APHCN-ASEANET mainly provides the technical capacity building program for the ASEAN plant protectionists.