PORCINE CYSTICERCOSIS IN PAPUA, INDONESIA VETERINARY ASPECTS OF PORCINE CYSTICERCOSIS IN JAYAWIJAYA REGENCY, PAPUA PROVINCE, INDONESIA Inriyanti Assa1, Fadjar Satrija2, Denny Widaya Lukman2, Nyoman Sadra Dharmawan3, Sri Murtini2 and Arius Togodly1 1Department of Epidemiology, Faculty of Public Health, Cenderawasih University, Papua; 2Department of Animal Disease and Veterinary Public Health, Faculty of Veterinary Medicine, Bogor Agricultural University (IPB), Bogor; 3Faculty of Veterinary Medicine, Udayana University, Bali, Indonesia Abstract. Taenia solium cysticercosis is a significant public health problem in Papua, Indonesia. Porcine cysticercosis is highly endemic in areas of Jayawijaya due to the strong tradition of pig husbandry. This study determined seroprevalence using monoclonal antibody-based sandwich enzyme-linked immunosorbent as- say (MoAb-ELISA) of porcine cysticercosis between October 2009 and June 2011 among 104 pigs in eight districts of Jayawijaya. The highest prevalence (93%) was in Asolokobal District and the lowest (6%) in Wamena Kota District, with an overall prevalence of 37%. Pigs in the Asolokobal District were highly vulnerable to porcine cysticercosis. In Jayawijaya, pigs are cooked in a traditional manner using hot stones, the temperature of which is 300°C, for 90 minutes resulting in an interior meat temperature of 60-90°C that is unable to kill cysticerci. Thus, it will be necessary to incorporate an anthropological approach to the system of pig husbandry and pork cooking practice if porcine cysticercosis is to be reduced in Jayawijaya. Keywords: porcine cysticercosis, sandwich-ELISA, seroprevalence, Indonesia INTRODUCTION infect brain and central nervous system (neurocysticercosis) or eyes (ocularcysti- Cysticercosis is a zoonotic disease cercosis) (Patel et al, 2011). Cysticercosis is caused by ingestion of eggs or proglotids a disease associated with poverty, indis- of tapeworm Taenia solium. As the de- criminate defecation and free roaming of finitive host of this parasite, humans can pigs (Rajshekhar et al, 2003). suffer from cysticercosis if they acciden- In Asia, there are three Taenia spp that tally ingest T. solium eggs. Cysticercosis in could infect humans, namely, T. asiatica, humans can be severe should the parasite T. saginata and T. solium (Ito et al, 2003; Correspondence: Inriyanti Assa, Department Okamoto et al, 2007). While human cysti- of Epidemiology, Faculty of Public Health, cercosis is caused only by T. solium, pigs Cenderawasih University, Kampus Abepura, are the intermediate hosts of T. asiatica and Jayapura, Papua 99351, Indonesia. T. solium, and many genetic similarities E-mail: [email protected] have been found between T. asiatica and Vol 49 No. 3 May 2018 375 SOUTHEAST ASIAN J TROP MED PUBLIC HEALTH T. saginata (Puchades and Fuentes, 2000; reinforce courage in battle (Department Ito et al, 2005; Anantaphruti et al, 2007; of Education, 1993). Because pigs are cul- Okamoto et al, 2007). turally important and an important food Cysticercosis/taeniosis can be found source for the indigenous community, in seven provinces of Indonesia, namely, efforts to improve the safety of pig meat Bali, Lampung, North Sulawesi, North consumption and to prevent the transmis- Sumatra, Papua, Southeast Sulawesi, and sion of foodborne illnesses to humans are West Kalimantan (Margono et al, 2006). needed. The disease is endemic in Bali (T. saginata Research on cysticercosis/taeniosis and T. solium), North Sumatra (T. asiatica), in this region has mostly been conducted and Papua (T. solium). The first case of in humans (Handali et al, 1997; Margono cysticercosis/taeniosis in Papua was re- et al, 2006; Salim et al, 2009). Recent sero- ported in 1972 following the discovery of epidemiological survey showed a high T. solium proglottids and eggs during a prevalence of human cysticercosis/taenio- routine examination at Enarotali Hospital, sis, 20.8% based on rT24H (cysticercosis Nabire Regency in the Central Highlands antigen) enzyme-linked immunoelectro- (Margono et al, 2006). Since then the dis- transfer blot (EITB) and 7% for taeniosis ease has spread to other regions including based on rES33 (taeniosis antigen) EITB Paniai and Jayawijaya. The spread of the in Jayawijaya (Salim et al, 2009). These disease is associated with the absence results indicated that cysticercosis and of latrines and poor personal hygiene, taeniosis are still prevalent and their infec- consumption of feces by straying pigs, tion rates have not changed since the first and cultural practice of consuming raw report 35 years ago. or undercooked meat. Suroso et al (2006) The study of cysticercosis in pigs as and Margono et al (2006) reported the a source of human infection is very lim- suspicion that cysticerci in pork cooked ited. The seroprevalence of cysticercosis using “bakar batu” (a stone cooking pit), in pigs in Jayawijaya from 1998 to 1999 a traditional cooking method of Papua ranges from 8.5 to 70.4% based on copro- involving pre-heated stones, is a source ELISA of pigs (Ito et al, 2003, Wandra et al, of infection in humans. 2007a,b). Maitindom (2008) conducting Wamena, the capital city of Jayawi- a post-mortem study in pigs sold at the jaya, is derived from the word “wam”, Jibama Market, Wamena, Jayawijaya meaning pigs. This suggests that pig is Regency reported a 77.1% prevalence of an important animal for the autochtho- swine cysticercosis. Thus, it is important nous population, the Dani ethnic group. to study some of veterinary aspects of por- The main types of pigs kept in Wamena cine cysticercosis in Jayawijaya Regency, are local breeds and some crossed with Papua, Indonesia to determine the sero- imported pigs, such as those of the large prevalence of porcine cysticercosis and white variety (Kataren, 2008). Various cer- pig farm management in the districts of emonies, performed in the public sphere, the Regency. In addition, we also carried involve pigs, and religious ceremonies out a measurement of pork meat tempera- of the Dani focus on either welfare or ture traditionally cooked using “bakar warfare ceremonies were held in the batu” to determine risk associated with past to invoke the tribe’s welfare and to this cultural practice. 376 Vol 49 No. 3 May 2018 PORCINE CYSTICERCOSIS IN PAPUA, INDONESIA MATERIALS AND METHODS phosphate-buffered saline (PBS) contain- ing 0.05% Tween 20 (washing buffer), fol- Study area lowed by incubating with 150 μl of block- The study was conducted in Jayawi- ing buffer (NBCS, Antwerp, Belgium) for jaya Regency of Papua Province, Indone- 15 minutes at 37°C with shaking. Then 100 sia (138°30’ - 139°40’ East and 3°45’ - 4°20› μl aliquots of treated serum were added South), and an altitude of 1,724 m above to the wells, incubated for 15 minutes at sea level. Data were collected from eight 37°C with shaking and then washed five [Asolokobal, Assologaima, Bolakme, times with washing buffer. Aliquots of Homhom, Hubikosi, Kurulu, Musatfak, 100 μl of detecting biotinylated MoAb and Wamena Kota (the capital city)] out of (B60H8A4 (Institute of Tropical Medicine, 11 districts in the Regency based on their Antwerp, Belgium) and 1.25 μg/ml block- accessibility. The study was carried out ing buffer were added and the plate was during October 2009 – June 2011. incubated as described above, followed by washing five times with washing buffer. Permit for the research was obtained Then 100 μl aliquot of orthophenylenedi- from the Livestock Service Office, Jayawi- amine (Sigma, St Louis, MO) [one table jaya Regency. spoon dissolved in 10 ml of aquabidest Monoclonal antibody-based sandwich en- (Ikapharmindo putramas, Jakarta, In- zyme-linked immunosorbent assay (MoAb- donesia)], 2.5 μl of H2O2 and 100 μl of ELISA) detection of porcine cysticercosis peroxidase-labelled streptavidin (Sigma) Blood serum samples were obtained (1:10,000 in blocking buffer) were added from 104 pigs [Asolokobal (n = 14), As- and incubated for 15 minutes at the 30°C sologaima (n = 22), Bolakme (n = 6), Hom- in the dark. The reaction was terminated hom (n = 11), Hubikosi (n = 7), Kurulu (n by the addition of 50 μl of 8 M H2SO4 and = 23), Musatfak (n = 4), and Wamena Kota absorbance measured at 492 and 655 nm (n = 17)] and 2 ml aliquots were stored at (Microplate Reader, Benchmark, Bio Rad, -20°C until used. MoAb-ELISA was per- London, UK). Appropriate positive and formed as previously described (Dorny et negative control samples were included al, 2004). In brief, 150 μl aliquot of serum in the assay. was incubated with an equal volume of Pig farm management survey 5% trichloroacetic acid for 20 minutes at Replies to a questionnaire was ob- room temperature, centrifuged 12,000g for tained from 56 pig owners in the study 9 minutes, and 150 μl aliquot of superna- areas to collect information on pig farm tant then was added to 75 μl of neutraliz- management. Interviews were conducted ing buffer (0.156 M carbonate/bicarbonate by Jayawijaya Regency Livestock Services pH 10). A 100 μl aliquot of capture anti- officers using local Dani language. MoAb (B158C11A10; Institute of Tropical Medicine, Antwerp, Belgium) and 5 µl/ Heated stone meat cooking technique ml 0.5 M bicarbonate buffer pH 9.8 were The “bakar batu” cooking technique applied to wells of a 96-well microtiter consists of heated stones to roast pork plate pre-treated with 100 µl of coating and vegetables. These large hot stones are buffer (0.05 M bicarbonate buffer pH 9.8) placed into a pit (about 70 cm deep) using and incubated for 30 minutes at 37°C a special clamp (“apando”) and then cov- with shaking. Wells were washed with ered with weeds (Fig 1). The raw pork was Vol 49 No. 3 May 2018 377 SOUTHEAST ASIAN J TROP MED PUBLIC HEALTH covered with grass and tied with rattans to spondents with a pig pen, 88% had a semi- prevent steam generated from the heat of permanent enclosure, 2% a permanent the hot stone from evaporating.