Journal of Global Pharma Technology

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Articles Journal Content Current Issue (/index.php/JGPT/issue/current) Search Past Issues Surveillance and Monitoring Brucellosis at (/index.php/jgpt/issue/archive) Articles in Press Denpasar Veterinary Center Working Area in the Search Scope (/index.php/JGPT/issue/archive) 2016 (http://www.jgpt.co.inA/lilndex.php/jgpt/article/view/3 (http://www.jgpt.co.in/index.php/jgpt/article/view/3041) JGPT Dedicated In Search Honour Of Great Suharsono H Scientist The Polymorphism of rs2549782 gene Endoplasmic Browse

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Journal of Global Pharma Technology

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Scope Journal of Global Pharma Technology (JGPT) is peer reviewed, Monthly open access Journal. JGPT publishes original research work that contributes signicantly to further the scientic knowledge in pharmacy and pharmaceutical sciences (Pharmaceutical Technology, Pharmacognosy, Natural Product Research, Pharmaceutics, Novel Drug Delivery, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics, Pharmacoeconomics). Research outcomes from medical sciences/case study and biotechnology of pharmaceutical interest are also considered. JGPT publishes original research work either as an Original Article or as a Short Communication. Review articles on current topic under mentioned scopes are also considered for publication.

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Indonesia. 2. Department of Internal Medicine, Wangaya Hospital, Denpasar, Bali, Indonesia. 3. Denpasar Veterinary Center, Directorate General of Animal Husbandry and Animal Health, Ministry of Agriculture.

Abstract

Background: The situation of Brucellosis in the working area of the Denpasar Veterinary Center (BBVet) varies between provinces. The provinces of Bali and NTB have been declared free of Brucellosis. However, specifically in NTT, only Sumba Island was declared free of Brucellosis. The situation of Brucellosis in NTT Province, on the island of Timor, Belu Regency and TTU is a heavily infected area of brucellosis with a prevalence of > 2%, while other islands have an unknown prevalence. One Brucellosis reactor found in Ende District in 2006. Continuous surveillance was carried out as an early detection step to keep the area free. Objectives: Brucellosis and monitor the possibility of entry/emergence of new reactors in the region, and to determine the prevalence of Brucellosis in areas that are not yet free of Brucellosis. Methodology: The samples tested were samples received by the BBVet Denpasar Bacteriology laboratory during 2016. The samples were tested using the Rose Bengal Plate Test (RBPT) method if positive continued with the Complement Fixation Test (CFT) test. Results: A positive sample of CFT expressed as a Brucellosis reactor. The test results of 4,602 serum samples from Bali Province and 2,504 serum samples from NTB Province were all negative for Brucella antibodies. Whereas serum samples from NTT Province were 1,533 samples, three positive samples were CUC brucellosis; namely, two (2) positive samples from 175 serum samples came from Malacca District and one (1) positive sample from 73 serum samples came from North Central Timor District ( TTU). Whereas samples from the districts of Alor, Ende, Lembata, Manggarai, West Manggarai, East Manggarai, Nagekeo, Ngada, RotenDao, Southwest Sumba, West Sumba, East Sumba, East Timor Tengah (TTS), and Kupang City, Conclusion: all negative. Based on these data it can be concluded that the Provinces of Bali and NTB are still free of Brucellosis.

Keywords: Brucellosis, BPT, CFT, Bali, NTB. NTT. Preliminary Brucellosis in cattle is usually caused by 7/2002 for Lombok Island in Nusa Tenggara Brucella abortion, which is one of the Barat (NTB) Province, and Minister of economically significant diseases because it is Agriculture Decree No. 97 / Kits / PO.660 / zoonotic (transmitted to humans). Besides, B. 2/2006 for Sumbawa Island in NTB province. abortion can be used in bioterrorist attacks However, specifically in Nusa Tenggara [1].Brucellosis is one of 22 strategic infectious Timur (NTT) Province, only Sumba Island animal diseases in Indonesia, is zoonotic was declared free of Brucellosis by Decree of (contagious to humans) and is a disease that the Minister of Agriculture No. 52 / Kits / is difficult to treat. PD.630 / 1/2015 dated 19 January 2015.

Bali Island, Lombok Island, and Sumbawa The situation of Brucellosis in NTT Province Island have been declared as Brucellosis-free varies among existing islands. On the island areas by the Minister of Agriculture of the of Timor, Belu and TTU districts are heavily Republic of Indonesia with a Minister of infected with brucellosis with a prevalence of Agriculture Decree No. 443 / Kits / TN.540 / >2%, while other islands have an unknown 7/2002 for Bali Island, Minister of prevalence. Brucellosis has been found in Agriculture Decree No. 444 / Kits / TN.540 / several districts on Flores Island such as

Ende Regency in 2002 [2], Sikka Regency in  Shake for 4 minutes until homogeneous 1996. Continuous surveillance is carried out using a rotary agglutination and read the as an early detection step to maintain a results. region free of Brucellosis and monitor the possibility of the entry/emergence of new The CFT Test Procedure is as follows: reactors in the region, and to determine the  Enter the serum to be tested onto each 50µl prevalence of Brucellosis in areas that are hole from the serum 1A hole for sample no. not yet free of Brucellosis. For this reason, 1, to the ser um 10A hole for sample no. 10, the Denpasar Veterinary Center has negative control serum 11A hole, hole 12B conducted surveillance in the work areas of for the positive serum control. The plate is Bali, West Nusa Tenggara, and East Nusa water bathed for 30 minutes to be Tenggara Provinces. activated. (all serums including positive Materials and Methods and negative controls) Theory  Add 25µl CFT buffer in holes B1 - B12 to holes H1 - H12 (holes A1 - A12 do not add In the surveillance of brucellosis in the CFT buffer) working area of the 2016 Denpasar Veterinary Center, the ingredients used were  Dilute Serum: serially, take 25 µl from hole Brucella abortus RBPT and CFT antigens, A1-12 to B1-12 to hole H1-12 supplements, hemolysin, sheep blood cells, cut buffers, and the tools used were  Add the Antigen (depending on the microplate, WHO plate, pipette, incubator, available antigen titer) 25 µl to hole C1-12 rotary agglutination, and rotary to hole H1-12. In holes, A1-12 and B1-12 as agglutination. anti-complement control added 25µl CFT buffer (to equalise volume) Method The samples tested were samples received by  Add Complement (depending on available the BBVet Denpasar Bacteriology laboratory complement titers) 25µl of all plate holes during 2016. The samples were tested using from A to H, incubation at 37oC for 30 the Rose Bengal Plate Test (RBPT) method if minutes. positive continued with the Complement  Add to all 25µl plate hole cells, then shaker Fixation Test (CFT) test [3]. for 45 minutes. The RBPT test procedure is as follows:  Leave it for a while and do the reading.  Serum samples removed from the freezer, Results and RBT brucella antigens are removed from the refrigerator and allow a few Test Results 4,602 serum samples from Bali minutes at room temperature. Province and 2,504 serum samples from NTB Province were all Brucellosis negative. While  The serum to be tested is taken with a the test results of 1,533 samples from NTT Pasteur pipette and drops on the WHO Province, three positive samples of plate (80 holes), in holes number 1 to Brucellosis by CFT were three samples from number 78 for the tested serum. Positive Malacca Regency and 1 sample from North serum control dropped on hole number 80, Central Timor Regency. Complete results as after that drops of RBT brucella antigens presented in Tables 1, 2 and 3. (25μl) were equal in all holes.

Table 1: Brucellosis Serology Test Results in Bali Province Districts Number of Samples Total Positive Brucellosis Badung 522 0 Bangli 385 0 Buleleng 616 0 Denpasar 193 0 Gianyar 641 0 Jembrana 1045 0 Karangasem 400 0 Klungkung 350 0 Tabanan 450 0 Amount 4602 0

Table 2: Brucellosis Serology Test Results in NTB Province Districts Number of Samples Total Positive Brucellosis Bima 287 0 Kota Bima 0 0 Dompu 651 0 Sumbawa 377 0 Sumbawa Barat 50 0 Island Sumbawa 1365 0 Lombok Barat 166 0 Lombok Tengah 156 0 Lombok Timur 417 0 Lombok Utara 277 0 Mataram 123 0 Island Lombok 1139 0 Amount NTB 2504 0

Table 3: Results of the Brucellosis Serological Test of NTT Province Districts Number of Samples Total Positive Brucellosis Alor 100 0 Ende 25 0 Kota Kupang 178 0 Kupang 108 0 Lembata 50 0 Malaka 175 2 (1,14%) Manggarai 50 0 Manggarai Barat 50 0 Manggarai Timur 50 0 Nagekeo 93 0 Ngada 65 0 Rote Ndao 28 0 SBD 80 0 Sumba Barat 155 0 Sumba Timur 140 0 TTU 73 1 (1,37%) TTS 113 0 Amount 1533

Discussion Brucellosis. In 2016, Brucellosis in East Timor found in Malacca District (1.14%) and In the working area of the Denpasar TTU (1.37%) as it is known that East Timor Veterinary Center, Bali Island has been is a Brucellosis-infected area with a declared historically free of Brucellosis. prevalence of <2% in TTS, Kupang and Lombok Island has been released from Kupang City, and prevalence of> 2% in TTU Brucellosis since 2002 (Minister of and Belu Districts (including Malacca Agriculture Decree Number 444 / Kits / District which is a division of Belu Regency). TN.540 / 7/2002), through mass surveillance In Belu and TTU districts, a Brucellosis for three years and then followed by the vaccination program has been carried out release of Sumbawa Island in 2006 (Minister using the Brucella abortus strain S19 of Agriculture Decree Number: 97 / Kits / vaccine, so it is not known with certainty PO.660 / 2/2006), with the same pattern of whether the antibody came from the results liberation as Lombok Island [4]. of vaccination or natural infection. All reactors found within the liberation The prevalence of brucellosis in Kupang City period have been destroyed or forced to cut. and Kupang Regency is uncertain even Then following the island of Sumba was though the results of the 2016 sample testing declared free of brucellosis based on the are all negative. It needs to be further decision of the Minister of Agriculture No. 52 confirmed by adequate sampling according to / Kits / PD.630 / 1/2015 dated January 19, epidemiological rules so that the actual 2015. The results of testing 4,602 serum prevalence can be known because the results samples against Brucellosis in 2016 in the of the 2015 surveillance show that there are Bali province, all were negative for indications of an increase in reactor Brucellosis. As is the case for the province of prevalence in Kupang City. The results of the West Nusa Tenggara, of the 2,504 serum 2015 Brucellosis surveillance on Flores samples tested came from Sumbawa and Island in Sikka, Ngada, Nagekeo District Lombok, all were negative for Brucella were all negative, as well as in 2016 all antibodies. It indicates that until now the samples from Ende, Manggarai, West provinces of Bali and NTB are still free of

Manggarai, East Manggarai, Nagekeo, and Conclusions and Suggestions Ngada Districts were negative brucellosis. The prevalence of Brucellosis on Flores Conclusion Island is still very low, Brucellosis on Flores Based on the surveillance results above, it Island was reported in Ende District in 1 cow concluded that in 2006 [2] and the cow has conditionally slaughtered.  The Provinces of Bali and West Nusa Tenggara are still areas free of Brucellosis Based on the results of surveillance in several years on the island of Flores, the  More intensive surveillance is needed on possibility for eradication programs is East Timor to get a more accurate possible to carry out, before it becomes Brucellosis prevalence. spreading. Brucellosis in other Regencies in NTT Province such as Lembata Regency,  The Brucellosis eradication program on Saburaijua Regency, Rotendau Regency is Alor, Lembata, Flores and Rotendau Island still negative, but to be declared a is possible Brucellosis-free region a structured survey Suggestion needs to be carried out with a sample that meets epidemiological requirements and is Further surveillance is needed by taking carried out simultaneously and continuously samples that are more representative and and tightens livestock traffic. Inter-island. comply with epidemiological rules, and to get more accurate Brucellosis prevalence data on the Land of Timor [5, 6]. References 1. OIE (2009) Bovine Brucellosis. OIE Veterinary Bulletin, BPPV Regional VI Terrestrial Manual, 1-35. Denpasar, XVIII (68): 46-54. 2. Dartini dan Rince MB (2007) Deteksi Dini 5. Putra AAG, Ekaputra IGM, Semara Putra Reactor Brucellosis in Ende Regency and AAG, Dartini NL (1995) Prevalence and Ngada Regency, Veterinary Bulletin, Distribution of Reactor Brucellosis in the BBVet Denpasar. Nusa Tenggara Region in 1994-1995. BPPH Report Region VI Denpasar. 3. OIE (2016) Brucellosis (Brucella abortus, B.melitensis, and B.suis) (Infection with B. 6. Putra AAG (2001) Epidemiology Study abortus, B.melitensis, and B.suis). OIE and the economic impact of brucellosis on Terrestrial Manual. Chapter 2.1.4. farmers', regional and national income: With an emphasis on East Nusa Tenggara 4. Putra AAG, Arsanai NM, Dartini NL, Province, Veterinary Bulletin, XIII (58): 8- Semara Putra AAG, Rince MB (2006) 18. Final evaluation of eradicating brucellosis in cattle / buffalo on Sumbawa Island,