Activities of JICA Experts in DIC Subang

1 General Activities 　 1) Chief Advisor / Animal Health Administration 　  - We continue the diagnostic activity on the specimens collected from the diseased animals in the field and analyzed the data on the diseases. Therefore the diagnostic ability of the laboratories in Subang DIC has been enhanced gradually.     - The diagnostic ability of the staff in Pathology Laboratory was enhanced by obtaining the histopathological staining techniques including immuno-histopatjological staining technique transferred by JICA short-term expert. - The staff in Epidemiology and Bacteriology Laboratories started the surveillance of brucellosis in cattle. - The staff in Bacteriology Laboratory isolated mycoplasma from lung of chicken. It is the first findings in DIC Subang. 　 2) Veterinary Diagnosis / Epidemiology     - Active field surveys on poultry diseases were conducted by the DIC in both districts of Cianjur and Garut of West Java Province, Bekasi district of Jakarta, and districts of Cilegon and Tangerang of Banten Province. Major sampling of blood and cotton swab was made, including autopsy materials in a few cases, from 6 to 10 fowl flocks in respective districts.  Inspection on materials of cotton swabs and autopsy was made with chicken embryonated eggs (CEE) through a second passage. As a result, no AI (H5) virus was isolated from each district surveyed except Bekasi, of which materials are under inspection, and ND virus was isolated from one flock of Tangerang district. HI antibody inspection of 667 sera samples was made on AI (H5) and ND. The result showed that positive reactors of AI (H5) ranged from less 1 % to more 90 %, and ND positive reactors ranged from slightly above 10 % to more 90 % in cases of respective districts.     - Responding to disease outbreak report and request from Indramayu district, West Java, where breeding of fighting cock is prospering, the DIC, Subang investigated the outbreak cases in fighting cock breeding farms of a few sub districts. As a result, suspicion of Gumboro disease was denied, and infection of velogenic ND or HPAI was suspected because any flocks of fighting cocks had never been vaccinated to both diseases. Sampling of blood and cotton swab was made      in 13 flocks with autopsy materials of two cocks. Inspection on samples of cotton swabs and autopsy was made with chicken embryonated eggs (CEE) through a second passage. As a result,      no viruses of ND and AI were isolated from cotton swab materials, however, AI (H5) virus was isolated from one of two autopsy materials. While, the pathology section of the laboratory also found the AI virus in the histopathology section of the same cock through immunohistochemical technique. Inspection results of both sections came to an accord. The isolated AI (H5) virus showed typical influenza CPE on CEF cell culture and was suspected as HPAI virus. Although one flock seemed to be attacked by HPAI, causative agents in other flocks’ cases are still unknown. HI antibody inspection of ND and AI(H5) on samples of the flocks showed that positive reactors of AI (H5) accounted for more 20 % (14/63), while ND positive reactors for about 5 % only (4/63).  Leaving an appropriate interval, the DIC needs to survey again in the area to check seroconvert condition on ND/ AI (H5) in survival cocks. - 36 field isolates of AI (H5) virus were inspected about CPE formation ability on CEF cell culture. As a result, 34 isolates showed CPE formation, however remaining 2 isolates failed to make CPE. The 2 isolates will be submitted to re-test in the near future. -  3 cases for Rabies diagnosis request were applied in this April. FAT showed negative results in any case. Mouse inoculation tests were initiated, and mice are under observation.          Mouse inoculation test on one case for Rabies diagnosis, which was applied late in latest March, completed (observation for about one month). Although all the mice survived, the brains were tested with FAT for final confirmation. The result was duly negative. - Serological tests made on bovine diseases were as follows in April, A. Bovine sera: 196 samples collected through active service　 - BVD ELISA antibody：positive 88, negative108 - IBR ELISA antibody：positive175, negative 21 - EBL（enzootic bovine leucosis）ELISA antibody：positive195, negative 1, B. Bovine sera: 43 samples collected through active service　 　- IBR ELISA antibody：positive 13, negative 30 　- EBL ELISA antibody：positive 2, negative 41 C. Bovine sera: 188 samples collected through active service - EBL ELISA antibody：positive 5, negative 183 D. FMD monitoring Bovine sera:47 samples: all negative 3) Coordinator / Animal Health Information     - As part of pilot site activity, we will conduct brucellosis surveillance in West Java province, and the actual investigation has been started from this month. In 10 prefectures in West Java province, 110,000 head dairy cattle has been raised. DIC Subang is planning to take 6,480 diagnostic samples there. More than 40 times brucellosis surveillance will be conducted in 2012, but the experts should go together to the active surveillances and continue transferring techniques and knowledge especially about the biosafety and sampling method.     - Biosafety workshop was held in Bali for four days (Society for Biosafety Asia Pacific Ocean), and 2 counterpart personnel (Dr.Isrok & Dr.Aji) have joined in it. They will make presentations at the seminar in DIC Subang next month. - Related to the activity termination of Dr. Mikami (pathological diagnosis technology), we reported the accomplishment to DGLAHS and JICA. To improve the IHC techniques further, the acquisition of IHC techniques for diseases other than AI, and obtaining of the positive control will become the new issues. 4) Bacteriological Diagnosis     - The staff in Bacteriology Laboratory isolated mycoplasma from lung of chicken. The isolated strain produced “Film and Spot” and does not required NAD for growth. It is the first isolation of mycoplasma in DIC Subang. - We collected 10 specimens of feces from one chicken farms (#23) as the activity of active surveillance of DIC Subang. No salmonella were isolated from the specimens of feces. Accumulatively, we isolated 7 strains of salmonella from 219 fecal samples collected from 4 of 23 chicken farms.     - We cultivated bacteria from specimens (ear, blood and soil) collected from anthrax-suspected died cow and identified the bacteria. However, the isolated bacteria appeared not to be Bacillus anthracis.     - One long-term expert and 2 staff in Bacteriology Laboratory attended the training on diagnosis of brucellosis of cattle held in DIC Maros in South Sulawesi. 5) Pathological Diagnosis     - Diagnosis by immunohistological staining of AI, and special staining (Gram staining, Loffler’s methylene blue staining, Ziehl-Nielsen staining, PAS staining, Masson’s trichrome staining, Berlin blue staining) was techniques were transferred. - Necropsy training was performed by using dying experimental animal (guinea pig). Findings such as lack of vitamin C was not observed, some sort of infection was suspected. Currently, we are waiting for the results of the diagnosis in bacteriology lab. - Counterparts in pathology lab need to have more experience of slide reading which is the most important for pathological diagnosis. Continuous practice is essential. 2. Important issues in activities (1) Biotechnology section tested Hog Cholera (HC) virus (vaccine strain) with some primers available for detection of both viruses of HC and Bovine viral diarrhea (BVD). Although they gained the result on HC, they do not yet confirm its availability on BVD because BVD reference virus or gene extracted materials are not yet acquired. It is expected to acquire BVD reference virus. (2) By attending the biosafety conference and seeing the situation in other DIC, biosafety issues in DIC Subang has been recognized again. Also, short-term expert (Dr.Mikami) described the necessity of strengthening bio-security and biosafety management in his report, hence we have to continue to improve the situation further. 3. Results (1) The staff in Bacteriology Laboratory isolated mycoplasma from lung of chicken. It is the first isolation of mycoplasma in DIC Subang. (2) Biotechnology section tested Hog Cholera (HC) virus (vaccine strain) with some primers. As a result, expected PCR band was observed to confirm the availability of the primers. (3) C/Ps in pathology lab became capable to conduct six (6) new special staining method. 4. Planned activities in next month (1) Dispatch of short-term expert (Veterinary Epidemiology):  May 27–July 21 (2) Comparison test on Rabies ELISA antibody test kits. (3) Third country training about the diagnosis of brucellosis in Thailand:  May 29–June 7  Source: JICA Expert Report