Sains Malaysiana
52(5)(2023):
1383-1395
http://doi.org/10.17576/jsm-2023-5205-05
Detection of Leptospira
wolffii in Water and Soil on Livestock Farms in Kelantan After a Massive
Flood
(Pengesanan Leptospira wolffii dalam Air dan
Tanah di Ladang Ternakan di Kelantan Selepas Banjir Besar)
MOHAMMAD SABRI ABDUL RAHMAN1,3, SITI KHAIRANI BEJO1,*, ZUNITA ZAKARIA1, LATIFFAH HASSAN2 & MOHD AZRI ROSLAN1
1Department of
Veterinary Pathology & Microbiology, Faculty of Veterinary Medicine,
Universiti Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia
2Department of
Veterinary Laboratory Diagnostics, Faculty of Veterinary Medicine, Universiti
Putra Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia
3Department of
Veterinary Diagnostics, Faculty of Veterinary Medicine, Universiti Malaysia
Kelantan, 16100 Kota Bharu, Kelantan Darul Naim, Malaysia
Diserahkan: 13 Ogos 2022/Diterima:
28 April 2023
Abstract
Pathogenic leptospires cause leptospirosis in both animals and humans
around the world. Humans can be infected
by leptospires through direct contact with infected animals or indirect contact
with leptospires contaminated water or soil. Outbreaks of leptospirosis usually
occur after heavy rainfall and flooding, particularly in endemic areas. The aim
of this study was to detect the presence of leptospires in water and soil in
livestock farms after a massive flood in Kelantan. Samples were collected from
28 livestock farms in Kelantan, comprising 62 water samples and 62 soil
samples, respectively. The samples were
filtered and inoculated into a semisolid EMJH medium containing 5-FU for the
isolation of Leptospira spp. The Leptospira spp. isolates were then
identified using classical methods (1M NaCl, 8-azaguanine, and 13 °C), serology
(MAT), multiplex polymerase chain reaction (mPCR), and DNA sequencing. The
classical identification methods showed varying results and failed to
differentiate between pathogenic and non-pathogenic leptospires. MAT showed the
isolates reactions against serovars Autumnalis, Hebdomadis, Pyrogenes,
Bataviae, Patoc, and Wolffii. However, mPCR showed that all isolates were
non-pathogenic Leptospira spp.
Further identification using DNA sequencing found that all isolates were
identified as Leptospira wolffii, an
intermediate species of Leptospira.
The presence of L. wolffii in water
and soil provides evidence that this species is circulating in the environment
and could potentially transmit to humans and animals.
Keywords: Flood; Kelantan;
Leptospirosis; soil; water
Abstrak
Patogen Leptospira menyebabkan Leptospirosis pada haiwan dan manusia di seluruh dunia. Manusia
boleh dijangkiti bakteria ini melalui sentuhan langsung dengan haiwan yang
dijangkiti atau melalui sentuhan tidak langsung dengan air atau tanah yang
tercemar. Wabak leptospirosis biasanya berlaku selepas hujan lebat dan banjir,
terutamanya di kawasan endemik. Matlamat kajian ini adalah untuk mengesan
kehadiran Leptospira dalam air dan tanah di ladang ternakan selepas
banjir besar di Kelantan. Sampel diperoleh daripada 28 ladang ternakan di Kelantan
yang masing-masing terdiri daripada 62 sampel air dan 62 sampel tanah. Sampel
ditapis dan diinokulasi ke dalam medium EMJH separa pepejal yang mengandungi
5-FU untuk pemencilan Leptospira spp. Pencilan Leptospira spp.
kemudiannya dikenal pasti menggunakan kaedah klasik (1M NaCl, 8-azaguanine, dan
13°C), serologi (MAT), tindak balas berantai polimerase multipleks (mPCR) dan
penjujukan DNA. Kaedah pengesanan klasik memberikan hasil yang berlainan dan
gagal membezakan antara Leptospira patogen dan bukan patogen. MAT
menunjukkan tindak balas terpencil terhadap serovars Autumnalis, Hebdomadis,
Pyrogenes, Bataviae, Patoc, dan Wolffii. Walau bagaimanapun, mPCR menunjukkan
bahawa semua pencilan adalah Leptospira spp. yang tidak patogen.
Pengecaman lanjut menggunakan penjujukan DNA mendapati bahawa semua pencilan
dikenal pasti sebagai L. wolffii, spesies perantaraan Leptospira.
Kehadiran L. wolffii dalam air dan tanah menunjukkan bahawa spesies ini
beredar dalam alam sekitar dan berpotensi untuk menular kepada manusia dan
haiwan.
Kata kunci: Air; banjir;
Kelantan; Leptospirosis; tanah
RUJUKAN
Abd-Rahman, A.N., Hasnul-Hadi, N.H., Sun, Z.,
Thilakavathy, K. & Joseph, N. 2021. Regional prevalence of intermediate Leptospira spp. in humans: A meta-analysis. Pathogens 10: 943.
Ahmed, A., Grobush, M.P., Klatser, P. &
Hartskeerl, R.A. 2012. Molecular approaches in the detection and
characterization of Leptospira. Journal of Bacteriology &
Parasitology 3: 133.
Ahmed, S.A., Doblin, A.S., Musa, S., Hoe, C.H.,
Riadzi, M., Lau, K.L. & Tang, T.H. 2011. Rapid diagnosis of leptospirosis
by multiplex PCR. Malaysian Journal of Medical Sciences 19(3): 9-16.
Bomfim, M., Barbosa-Stancioli, E.F. & Koury, M.C.
2008. Detection of pathogenic leptospires in urine from naturally infected
cattle by nested PCR. Veterinary Journal 178(2): 251-256.
Fonseca, C., Teixeira, M.,
Romero, E.C., Tengan, F.M., da Silva, M.V. & Shikanai-Yasuda, M.A. 2006.
Leptospiral DNA detection for the diagnosis of human leptospirosis. Journal
of Infections 52(1): 15-22.
Hartskeerl, R.A., Smits, H.L., Korver, M., Goris, M.G.
& Terpstra, W.J. 2001. International Course on Laboratory Methods for
the Diagnosis of Leptospirosis.Netherlands: Royal Tropical
Institute Department of Biomedical Research.
Johnson, R.C. & Faine, S. 1984. Genus I. leptospira noguchi 1917. In Bergey’s Manual of Systematic
Bacteriology, edited by Krieg,
N.R. & Holt, J.G. Baltimore: Williams & Wilkins. pp.
62-67.
Johnson, R.C. & Gary, N.D. 1962. Nutrition of Leptospira
pomona. 1. A chemically defined substitute for rabbit serum ultrafiltrate. Journal
of Bacteriology 83(3): 668-672.
Kmety, E., Plesko, I., Bakoss, P. & Chorvath, B.
1966. Evaluation of methods for differentiating pathogenic and saprophytic
leptospiral strains. Annales des Sociétés Belges de Médecine Tropicale, de
Parasitologie, et de Mycologie 46: 111-122.
Kumar, S., Stecher, G. & Tamura, K. 2016. MEGA7:
Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular
Biology and Evolution 33: 1870-1874.
Laras, K., Cao, B.V., Bounlu, K., Nguyen, T.K.T.,
Olson, J.G., Thongchanh, S., Nguyen, V.A.T., Hoang, K.L., Punjabi, N., Ha,
B.K., Ung, S.A., Insisiengmay, S., Watts, D.M., Beecham, H.J. & Corwin,
A.L. 2002. The importance of Leptospirosis in Southeast Asia. The American
Journal of Tropical Medicine and Hygiene 67(3): 278-286.
Levett, P.N. 2004. Leptospirosis: A forgotten
zoonosis? Clinical & Applied Immunology Reviews 4: 435-448.
Lilenbaum, W., Varges, R., Brandão, F.Z., Cortez, A.,
de Souza, S.O., Brandão, P.E., Richtzenhain, L.J. & Vasconcellos, S.A.
2008. Detection of Leptospira spp. in semen and vaginal fluids of goats
and sheep by polymerase chain reaction. Theriogenology 69(7): 837-842.
Merien, F., Amouriaux, P., Perolat, P., Baranton, G.
& Saint Girons, I. 1992. Polymerase chain reaction for detection of Leptospira spp. in clinical samples. Journal of Clinical Microbiology 30:
2219-2224.
MOH. 2015. Epidemiology
and current situation of Leptospirosis in Malaysia. Dr. Zainudin AW. Persidangan Kesihatan
Persekitaran Pihak Berkuasa Tempatan 2015, 8-9 September 2015, WP
Labuan.
Mohd-Radi, M.F., Hashim, J.H., Jaafar, M.H., Hod, R.,
Ahmad, N., Mohammed Nawi, A., Baloch, G.M., Ismail, R. & Farakhin Ayub,
N.I. 2018. Leptospirosis outbreak after the 2014 major flooding event in
Kelantan, Malaysia: A spatial-temporal analysis. The American Journal of
Tropical Medicine and Hygiene 98(5): 1281-1295.
Noubade, R., Krishnamurthy, G., Murag, S., Venkatesha,
M. & Krishnappa, G. 2002. Differentiation of pathogenic and saprophytic
Leptospires by polymerase chain reaction. Indian Journal of Medical
Microbiology 20(1): 33-36.
OIE. 2014. Manual
of Diagnostic Tests and Vaccines for Terrestrial Animals. http://www.oie.int/manual-of-diagnostic-tests-and-vaccines-for-terrestrial-animals/. Accessed on 29 January 2017.
Ooteman, M.C., Vago, A.R. & Koury, M.C. 2006.
Evaluation of MAT, IgM ELISA and PCR methods for the diagnosis of human
leptospirosis. Journal of Microbiology Methods 65(2): 247-257.
Pui, C.F., Bilung, L.M., Apun,
K. & Su’ut, L. 2017a. Diversity of Leptospira spp. in rats and
environment from urban areas of Sarawak, Malaysia. Journal of Tropical
Medicine 2017: 1-8.
Pui, C.F., Bilung, L.M.,
Su’ut, L., Chong, Y.L. & Apun, K. 2017b. Detection of Leptospira spp. in selected national service training centres and paddy fields of Sarawak,
Malaysia using polymerase chain reaction technique. Tropical Agriculture
Science 40(1): 99-110.
Pui, C.F.,
Bilung, L.M., Su’ut, L. & Apun, K. 2015. Prevalence of Leptospira species in environment soil and water from national parks in Sarawak, Malaysia. Borneo Journal Resource Science and Technology 5(1): 49-57.
Sabri, A.R., Khairani-Bejo, S., Zunita, Z. &
Hassan, L. 2019. Molecular detection of Leptospira sp. in cattle and
goats in Kelantan, Malaysia after a massive flood using multiplex polymerase
chain reaction. Tropical Biomedicine 36(1): 165-171.
Slack, A.T., Kalambaheti, T., Symonds, M.L., Dohnt,
M.F., Galloway, R.L., Steigerwalt, A.G., Chaicumpa, W., Bunyaraksyotin, G.,
Craig, S., Harrower, B.J. & Smythe, L.D. 2008. Leptospira wolffii sp. nov., isolated from a human with suspected leptospirosis in Thailand. International
Journal of Systematic and Evolutionary Microbiology 58(10): 2305-2308.
Tansuphasiri, U., Chanchai, T., Duangporn, P. &
Charnchudhi, C. 2006. Duplex PCR-hybridization based detection of pathogenic Leptospira in environmental water samples obtained from endemic areas in northeast region
of Thailand. Southeast Asian Journal of Tropical Medicine Public Health 37(4): 729-741.
Wan-Yussof, W.N. 2015. Flood in Kelantan in 2014: An
Introspection. Report from Centre for Strategic Studies, the State of
Kelantan, Malaysia.
Zakeri, S., Khorami, N.,
Ganji, Z.F., Sepahian, N., Malmasi, A.A., Gouya, M.M., Djadid, N.D. 2010. Leptospira
wolffii, a potential new pathogenic Leptospira species detected in
human, sheep and dog. Infection, genetics and evolution. Journal of
Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases 10(2): 273-277.
*Pengarang untuk surat-menyurat; email: skhairani@upm.edu.my
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