Detection of microbial antigens in clinical samples can lead to rapid diagnosis of an infection and administration of appropriate therapeutics. A major barrier in diagnostics development is determining which of the potentially hundreds or thousands of antigens produced by a microbe are actually present in patient samples in detectable amounts against a background of innumerable host proteins. In this report, we describe a strategy, termed in vivo microbial antigen discovery (InMAD), that we used to identify circulating bacterial antigens. This technique starts with “InMAD serum,” which is filtered serum that has been harvested from BALB/c mice infected with a bacterial pathogen. The InMAD serum, which is free of whole bacterial cells, is used to immunize syngeneic BALB/c mice. The resulting “InMAD immune serum” contains antibodies specific for the soluble microbial antigens present in sera from the infected mice. The InMAD immune serum is then used to probe blots of bacterial lysates or bacterial proteome arrays. Bacterial antigens that are reactive with the InMAD immune serum are precisely the antigens to target in an antigen immunoassay. By employing InMAD, we identified multiple circulating antigens that are secreted or shed during infection using Burkholderia pseudomallei and Francisella tularensis as model organisms. Potential diagnostic targets identified by the InMAD approach included bacterial proteins, capsular polysaccharide, and lipopolysaccharide. The InMAD technique makes no assumptions other than immunogenicity and has the potential to be a broad discovery platform to identify diagnostic targets from microbial pathogens.

Bantul is one of leptospirosis endemic areas which has been reported the cases every year and the presence of rats is an important risk factor relating to leptospirosis incidence in Bantul. Therefore, the role of rats as reservoir was examined in three types of ecosystems: forest, non-forest, and coastal ecosystems. Rat trapping was carried out using 100 single livetraps which were distributed in 9 locations: 2 points in the forest, 5 points in the non-forest, and 2 points in the coastal ecosystem. The rats were identified and their kidneys were collected and preserved in 70% alcohol medium. Leptospira were detected in the kidney by using PCR method. A number of 196 rats were obtained during the study. Most of them were commensal rats obtained from settlements ecosystems. Rattus tanezumi was the dominant species in the three ecosystems, but the highest infection rate of leptospira pathogen was in R.norvegicus. Coastal ecosystems were contained more infected rats, where the main habitat were mangrove forests. Based on these results, rats have great potency in leptospirosis transmission in Bantul, especially in settlements and coastal areas although leptospirosis cases were reported rarely in study area. Nonetheless, awareness of transmission should be disseminated since the presence of pathogenic leptospira in rats is very high. Keywords: leptospirosis, rat, reservoir, ecosystem, Bantul Abstrak Kabupaten Bantul merupakan salah satu daerah endemis leptospirosis yang sejak lama melaporkan kasus ini setiap tahunnya dan keberadaan tikus merupakan faktor risiko penting yang mempengaruhi kejadian leptospirosis di Kabupaten Bantul. Oleh karena itu peranan tikus sebagai hewan reservoir diteliti di tiga jenis ekosistem yaitu ekosistem hutan, non-hutan, dan pesisir. Penangkapan tikus dilakukan menggunakan perangkap tunggal sebanyak 100 buah yang disebar di 9 titik lokasi, meliputi dua titik di ekosistem hutan, lima titik di ekosistem non-hutan, dan dua titik di ekosistem pesisir. Tikus tertangkap diidentifikasi kemudian organ ginjalnya dikoleksi dan diawetkan dalam medium alkohol 70%. Bakteri leptospira dideteksi pada specimen ginjal dengan metode PCR dari total 196 tikus diperoleh selama penelitian. Sebagian besar tikus diperoleh dari ekosistem dekat pemukiman dan semuanya merupakan tikus komensal. Rattus tanezumi merupakan spesies dominan di ketiga ekosistem, namun persentase infeksi leptospira patogen paling tinggi terdapat pada R. norvegicus. Ekosistem pesisir mengandung tikus terinfeksi leptospira patogenik paling tinggi, dengan habitat utama hutan mangrove. Berdasarkan hasil ini maka tikus berpotensi besar menularkan leptospirosis di Kabupaten Bantul, terutama di daerah pemukiman dan pesisir. Meskipun kasus leptospirosis di daerah penelitian sedikit sekali dilaporkan, namun kewaspadaan terhadap penularan perlu dilakukan sedini mungkin karena leptospira patogenik yang terkandung dalam tikus tergolong tinggi. Kata kunci: leptospirosis, tikus, reservoir, ekosistem, Bantul

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Leptospirosis is still a health problem in the word. Leptospirosis can be transmitted to humans trough contact with environment infected with pathogenic Leptospira. Efforts to control pathogenic Leptospira in the environment can be done one of them by disinfecting. Effective desinfectants for the control of pathogenic Leptospira include the active ingredients Calcium hypochlorite 60% and Sodium hypochlorite 5,25%. This study aims to determine the effectiveness of Calcium hypochlorite 60% and Sodium hypochlorite 5,25% against the life power of pathogenic Leptospira. This research is a laboratory study with pure experimental design. Serovar of pathogenic Leptospira used is Leptospira icterohaemorrhagiae. Leptospira icterohaemorrhagiae desnsity used as much as 5.7 x 106 Leptospira/ml. The concentrations of desinfectans and bacteria are: 5 ppm, 50 ppm, 500 ppm, 1000 ppm, 2000 ppm, 5000 ppm, 10.000 ppm, and 25.000 ppm. Observations are made every ten minutes and twenty minutes with three repeats. The results showed that Leptospira icterohaemorrhagiae could not survive when tested on 60% calcium hypochlorite and 5.25% sodium hypochlorite, respectively, at a concentration of 2000 ppm and a concentration of 1000 ppm. Statistical results showed there was a significant difference in the concentration of each disinfectant active ingredient. Thus, a disinfectans of chlorine with the active ingredient Calcium hypochlorite 60% and Sodium hypochlorite 5,25% effectively affects the viability of the Leptospira icterohaemorrhagiae. Keywords : Sodium Hypochlorite, Calcium Hypochlorite, Leptospira, Disinfectant Abstrak Leptospirosis masih menjadi masalah kesehatan di dunia. Leptospirosis dapat menular ke manusia lewat kontak dengan lingkungan yang mengandung Leptospira patogenik. Upaya pengendalian Leptospira patogenik di lingkungan dapat dilakukan salah satunya dengan pemberian disinfektan. Disinfektan yang efektif untuk pengendalian Leptospira patogenik diantaranya adalah bahan aktif kalsium hipoklorit 60% dan sodium hipoklorit 5,25%. Penelitian ini bertujuan mengetahui efektivitas bahan aktif kalsium hipoklorit 60% dan sodium hipoklorit 5,25% terhadap daya hidup Leptospira patogenik. Penelitian ini merupakan penelitian skala laboratorium dengan rancangan eksperimental murni. Serovar Leptospira patogenik yang digunakan adalah Leptospira icterohaemorrhagiae. Kepadatan Leptospira icterohaemorrhagiae yang digunakan sebanyak 5,7 x 106 Leptospira/ml. Konsentrasi campuran disinfektan dan bakteri yaitu: 5 ppm, 50 ppm, 500 ppm, 1000 ppm, 2000 ppm, 5000 ppm, 10.000 ppm, dan 25.000 ppm. Pengamatan dilakukan tiap 10 menit dan 20 menit dengan tiga kali ulangan. Hasil penelitian menunjukkan Leptospira icterohaemorrhagiae tidak dapat hidup saat diujikan ke kalsium hipoklorit 60% dan sodium hipoklorit 5,25% berturut-turut pada konsentrasi 2000 ppm dan konsentrasi 1000 ppm. Hasil statistik menunjukkan ada perbedaan signifikan dari besaran konsentrasi masing-masing bahan aktif disinfektan. Dengan demikian, disinfektan klorin dengan bahan aktif calcium hypochlorite 60% dan sodium hypochlorite 5,25% efektif memengaruhi daya hidup Leptospira icterohaemorrhagiae. Kata kunci: Sodium hipoklorit, Kalsium hipoklorit, Leptospira, Disinfektan