Detection of Leptospira spp. in kidney tissues isolated from rats in the Napu and Bada Highlands of Poso District, Central Sulawesi Province

Detection of Leptospira spp. in kidney tissues isolated from rats in the Napu and Bada Highlands of Poso District, Central Sulawesi Province

  • Gunawan Gunawan National Institute of Health and Research Development Unit Donggala Ministry of Health, Republic of Indonesia
  • Tri Wibawa
  • Mahardika Agus Wijayanti
  • Hayani Anastasia
Keywords: Leptospira, 16S rRNA, LipL32, PCR, Kidney’s Rat

Abstract

Leptospirosis is still a global health problem because it affects human health in rural and urban areas, both in industrialized and developing countries. The aim of the study was to detect Leptospira spp. bacteria in kidney tissues isolated from rats in the Napu and Bada Highlands of Poso District, Central Sulawesi Province. Kidneys sample from 63 rats were collected from Napu and Bada Highlands of Poso District, Central Sulawesi Province in MayJune 2018. Polymerase Chain Reaction (PCR) was used to detect Leptospira. The molecular characterizations were conducted based on the 16SrRNA and LipL32 genes. Data were analyzed descriptively to describe the presence of pathogenic Leptospira DNA. Analysis phylogenetic was performed using MEGA 6.2 software. A total of 63 rats was successfully
caught during the study consisting of males and female for 36 (57.1%) and 27 (42.9%), respectively. The species of rats were R. exulans, R. tanezumi, R. argentiventer, R. norvegicus,  M. Musculus, Paruromys dominator, Maxomys sp., and Rattus sp. The pathogenic of Leptospira DNA was detected in rats with R. argentiventer and Paruromys dominator
species using the 16S rRNA and LipL32 gene. Sample sequences using LipL32 target gene is a close similarity with L. interrogans serovar Hardjo, serovar Autumnalis, Lai, Icterohaemorrhagiae, Balico, Grippotyphosa, Mini, Canicola, Hebdomadis; L. noguchii serovar Pomona and L. kirschneri whereas the sample sequence using 16S rRNA target
gene showed similarity with L. interrogans serovar Canicola, Copenhagen, Autumnalis, Pyrogenes, Javanica, Icterohaemorrhagiae, Manilae, Bratislava, Linhae, Hebdomadis, and L. kirschneri serovar Grippotyphosa. The PCR method with the target gene 16SrRNA and LipL32 are able to detect Leptospira spp. in rats R. argentiventer and P. dominator species

Keywords: Leptospira, 16S rRNA, LipL32, PCR, Kidney’s Rat

 

Leptospirosis masih merupakan masalah kesehatan global karena mempengaruhi
kesehatan manusia di daerah pedesaan dan perkotaan, baik di negara industri maupun
mnegara berkembang. Tujuan penelitian adalah untuk mendeteksi bakteri Leptospira spp di
jaringan ginjal dari tikus di Dataran Tingi Napu dan Bada Kabupaten Poso, Provinsi
Sulawesi Tengah. Ginjal tikus sebanyak 63 sampel dikoleksi dari Dataran Tinggi Napu dan
Bada Kabupaten Poso, Provinsi Sulawesi Tengah pada bulan Mei – Juni 2018. PCR digunakan
untuk mendeteksi Leptospira. Karakterisasi molekuler dilakukan berdasarkan gen 16SrRNA
dan LipL32. Data dianalisis secara deskriptif untuk menggambarkan keberadaaN
Leptospira yang patogenik. Analisis filogenetik dilakukan dengan menggunakan perangkat
lunak Mega 6.2. Sebanyak 63 tikus berhasil ditangkap selama penelitian yang terdiri dari
jantan dan betina, masing masing 36 ekor (75,1%) dan 27 ekor (42,9%). Spesies tikus adalah
R. exulans, R. tanezumi, R. argentiventer, R. norvegicus, M. Musculus, Paruromys dominator,
Maxomys sp, dan Rattus sp. DNA Leptospira patogenik terdeteksi pada tikus dengan spesies
R. argentiventer dan Paruromys dominator menggunakan gen 16SrRNA dan LipL32 Sekuen
sampel dengan target gen LipL32 menunjukkan kesamaan dengan L. interrogans serovar
Hardjo, serovar Autumnalis, Lai, Icterohaemorrhagiae, Balico, Grippotyphosa, Mini,
Canicola, Hebdomadis; L. noguchii serovar Pomona dan L. kirschneri. Sedangkan sekuen
sampel dengan target gen 16S rRNA menunjukkan kesamaan dengan L. interrogans serovar
Canicola,Copenhagen, Autumnalis, Pyrogenes, Javanica, Icterohaemorrhagiae, Manilae,
Bratislava, Linhae, Hebdomadis, dan L. kirschneri serovar Grippotyphosa. Metode PCR
dengan target gen 16SrRNA dan LipL32 mampu mendeteksi Leptospira spp. pada tikus
dengan spesies R. argentiventer dan P. dominator.

Kata kunci: Leptospira,  16S rRNA, LipL32,  PCR,  Ginjal Tikus

References

Paixão S, Alves-martin MF, Michely S, et al. Serology, isolation, and molecular detection of Leptospira spp. from the tissues and blood of rats captured in a wild animal preservation centre in Brazil. Prev Vet Med. 2014;115:69-73. doi:10.1016/j.prevetmed.2014.03.016

Vitale M, Agnello S, Chetta M, et al. Human leptospirosis cases in Palermo Italy. The role of rodents and climate. J Infect Public Health. 2017. doi:10.1016/j.jiph.2017.07.02

Latifah I, Abdul Halim A, Rahmat M, et al. Isolation by culture and PCR identiication of Lipl32 gene of pathogenic Leptospira spp. In wild rats of Kuala Lumpur. Malays J Pathol. 2017;39(2):161-166.

Cohen J, Powderly W, Opal SM. Infectious Disease. 4 th. Elsevier Limited. All right reserved; 2016.

Mulyono A, Ristiyanto, Rahardianingtyas E, Putro DBW, Joharina AS. Prevalensi dan Identiikasi Leptospira Patogenik Pada Tikus. Vektora. 2016;8(Nomor 1):31-40

Ko AI, Goarant C, Picardeau M. Leptospira: The Dawn of the Molecular Genetics Era for an Emerging Zoonotic Pathogen. Nat Rev M i c r o b i o l . 2 0 0 9 ; 7 ( 1 0 ): 7 3 6 - 7 4 7 . doi:10.1038/nrmicro2208.Leptospira

Picardeau M. Virulence of the zoonotic agent of leptospirosis: Still terra incognita? Nat Rev M i c r o b i o l . 2 0 1 7 ; 1 5 ( 5 ): 2 9 7 - 3 0 7 . doi:10.1038/nrmicro.2017.5

Rinanda. T. ANALISIS SEKUENSING 16S rRNA DI BIDANG MIKROBIOLOGI. J Kedokt Syiah Kuala. 2011;11 Nomer 3:172-177

Himsworth CG, Bidulka J, Parsons KL, et al. Ecology of Leptospira interrogans in Norway Rats ( Rattus norvegicus ) in an Inner-City Neighborhood of Vancouver , Canada. PLoS N e g l T r o p D i s . 2 0 1 3 ; 7 ( 6 ) . doi:10.1371/journal.pntd.0002270

Agudelo-lo P, Thiry D, Romero-vivas CME, Cuello-pe M, Levett PN, Falconar AKI. CrossSectional Study of Leptospira Seroprevalence in Humans , Rats , Mice , and Dogs in a Main Tropical Sea-Port City. Am Soc Trop Med Hyg. 2 0 1 3 ; 8 8 ( 1 ) : 1 7 8 - 1 8 3 . doi:10.4269/ajtmh.2012.12-0232

Ristiyanto, Wibawa T, Budihar ta S, Supargiono. Prevalensi Tikus Terinfeksi Leptospira interogans. Vektora. 2015;7 nomer 2:85-92.

Vieira AS, Narduche L, Martins G, et al. Detection of wild animals as carriers of Leptospira by PCR in the. Acta Trop. 2 0 1 6 ; 1 6 3 : 8 7 - 8 9 . doi:10.1016/j.actatropica.2016.08.001

WHO. Leptospirosis situation in the WHO South-East Asia Region. 2007

Iwasaki H, Chagan-yasutan H, Susan P, et al. Combined antibody and DNA detection for early diagnosis of leptospirosis after a disaster. Diagn Microbiol Infect Dis. 2 0 1 6 ; 8 4 ( 4 ) : 2 8 7 - 2 9 1 . doi:10.1016/j.diagmicrobio.2016.01.001

B2P2VRP. RISET KHUSUS VEKTOR DAN RESERVOIR PENYAKIT (RIKHUS VEKTORA), Laporan Provinsi Sulawesi Tengah. 2015.

B2P2VRP. Sekapur Sirih Riset Khusus Vektor dan Reservoir Penyakit (Rikhus Vektora). 2014.

BPS. Propinsi Sulawesi Tengah Dalam Angka 2017.; 2017.

Widjajanti W, Hayani Anastasia, Rosmini, Veridiana NN, Tri W, Yuana. Kewaspadaan Dini Kasus Leptospirosis. Vektora. 2017:59-68.

Promega. Techinal Manual Wizard ®Genomic DNA Puriication Kit Wizard ® Genomic DNA Puriication Kit. Promega Corp All Rights Reserv. 2017:10-11.

Cosson JF, Picardeau M, Mielcarek M, et al. Epidemiology of Leptospira Transmitted by Rodents in Southeast Asia. PLoS Negl Trop D i s . 2 0 1 4 ; 8 ( 6 ) . doi:10.1371/journal.pntd.0002902

Astuti DR. Keefektifan Rodentisida Racun Kronis Generasi II Terhadap Keberhasilan Penangkapan Tikus. J Kesehat Masy. 2013;8(16):183-189.

Priyambodo. Pengendalian Hama Tikus Terpadu. Jakarta: Penebar : Swadaya Jakarta; 2003.

Mauron C, Goarant C, Perez J, Brescia F. Ro d e n t Ab u n d a n c e Dy n a m i c s a n d Leptospirosis Carriage in an Area of HyperEndemicity in New Caledonia. PLoS Negl Trop D i s . 2 0 1 1 ; 5 ( 1 0 ) . doi:10.1371/journal.pntd.0001361

Nurisa I, Ristiyanto. Penyakit Bersumber Rodensia (Tikus dan Mencit) di Indonesia. J Ekol Kesehat. 2005;4(no 3):308-3019.

Kudo Y, Vansith K, Rin E, et al. Molecular Epidemiological Survey of Leptospira Infection of Wild Rodents in the Urban Settlement of Cambodia. Vector Borne Zoonoti c Dis. 2018;18(3):144-150. doi:10.1089/vbz.2017.2198

Haake DA, Chao G, Zuerner RL, et al. The Leptospiral Major Outer Membrane Protein LipL32 Is a Lipoprotein Expressed during Mammalian Infection. 2000;68(4):2276- 2285.

Azhari NN, Alia SN, Joseph N, et al. Molecular characterization of pathogenic Leptospira sp. in samll mammals captured from the human leptospirosis suspected areas of Selangor state, Malaysia. Acta Trop. 2018:1-34. doi:10.1016/j.actatropica.2018.08.020

Shukla J, Tuteja U, Batra H V. 16S rRNA PCR For Differentiation Of Pathogenic and non Pathogenic Leptospira Isolates. Indian J Med Microbiol. 2003;21(21):25-30.

Gokmen TG, Soyal A, Kalayci Y, Onlen C, Koksal F. COMPARISON OF 16S rRNA-PCRRFLP , LipL32-PCR AND OmpL1-PCR METHODS IN THE DI AGNOSIS OF LEPTOSPIROSIS. Rev Inst Med Trop Sao Paulo.

0 1 6 ; 5 8 : 6 4 ( 1 ) : 2 - 7 . doi:http://dx.doi.org/10.1590/S1678-9946201658064 ORIGINAL

Sumanta H, Wibawa T, Hadisusanto S, Nuryati A, Kusnanto H. Genetic Variation of Leptospira Isolated From Rats Catched in Yogyakarta Indonesia. Asian Pac J Trop Med. 2 0 1 5 ; 8 ( 9 ) : 7 1 0 - 7 1 3 . doi:10.1016/j.apjtm.2015.07.029

Picardeau M. Virulence of the zoonotic agent of Leptospirosis : still terra inognita? Nat Publ Gr. 2017. doi:10.1038/nrmicro.2017.5

Mohammed H, Nozha C, Hakim K, Abdelaziz F, Rekia B. Leptopsira : Classiication and P a t h o g e n e s i s . 2 0 1 1 ; 2 ( 6 ) : 6 - 9 . doi:10.4172/2155-9597.1000120

Anonim. Leptospirosis. Leptospirosis August 2013 At http://www.cfsph.iastate.edu/DiseaseInfo/factsheets.php. 2013:1-10.

I Wayan Suardana. Buku Ajar Zoonosis Penyakit Menular Dari Hewan Ke Manusia. PT Kanisius; 2016.

Published
2020-06-02
How to Cite
Gunawan, G., Wibawa, T., Wijayanti, M., & Anastasia, H. (2020). Detection of Leptospira spp. in kidney tissues isolated from rats in the Napu and Bada Highlands of Poso District, Central Sulawesi Province. Jurnal Vektor Penyakit, 14(1), 17-26. https://doi.org/10.22435/vektorp.v14i1.1965
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