Multilocus sequence typing reveals diverse known and novel genotypes of Leptospira spp. circulating in Sri Lanka

Karunanayake, Lilani; Gamage, Chandika D.; Gunasekara, Chandima P.; Silva, Sajiv De; Izumiya, Hidemasa; Morita, Masatomo; Muthusinghe, Devinda S.; Yoshimatsu, Kumiko; Niloofa, Roshan; Karunanayake, Panduka; Uluwattage, Wimalasiri; Ohnishi, Makoto; Koizumi, Nobuo

doi:10.1371/journal.pntd.0008573

Cited by 6 publications

(4 citation statements)

References 32 publications

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“…Genomic DNA was extracted from 0.5 mL cultures of the isolates using the DNeasy Blood and Tissue Kit (QIAGEN, Hilden, Germany). The partial flaB gene was amplified with Takara EX Taq Hot Start version (Takara Bio, Japan) using the primer set L- flaB -F1 and L- flaB -R1 as previously described [ 15 ]. The polymerase chain reaction (PCR) products were purified with ExoSAP-IT Express PCR Cleanup Reagents (Applied Biosystems, Foster City, CA, USA) according to the manufacturer’s instructions.…”

Section: Methodsmentioning

confidence: 99%

“…In contrast to the above specific serovar-animal association, multiple serovars/serogroups in a single animal species have been observed in small Indian mongoose or rats [1,8]. Recently, molecular typing methods such as multilocus sequencing typing (MLST) and multiple-locus variable-number tandem repeat analysis (MLVA) have been demonstrated to be useful tools for identifying Leptospira serovars/serogroups or discriminating strains within the same serovar [9][10][11][12][13][14][15]. These molecular typing methods indicated an association between virulence and specific Leptospira genotypes [12][13][14]16,17].…”

Section: Introductionmentioning

confidence: 99%

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Molecular epidemiology of Leptospira spp. among wild mammals and a dog in Amami Oshima Island, Japan

Shinya¹,

Muraoka

Negishi³

et al. 2021

PLoS ONE

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Leptospirosis is a worldwide zoonosis caused by the pathogenic Leptospira spp. Canine and human leptospirosis sometimes occur on Amami Oshima Island, located in the Nansei Archipelago, southwestern Japan; however, information on the causative Leptospira spp. on this island is quite limited. This study aimed to investigate the molecular and serological characteristics of Leptospira spp. isolated from wild animals and a dog in Amami Oshima Island. We obtained seven Leptospira strains by culturing kidney tissues of wild animals, such as black rats (2), wild boars (3), and rabbit (1) as well as blood from a symptomatic dog. Using flaB sequencing and microscopic agglutination test with antisera for 18 serovars, the isolates were identified as Leptospira borgpetersenii serogroups Javanica (black rat), L. interrogans serogroup Australis (black rat and dog), and L. interrogans serogroup Hebdomadis (wild boar and rabbit). The sequence type (ST) of L. borgpetersenii serogroup Javanica was determined to be ST143 via multilocus sequence typing (MLST) using seven housekeeping genes. For L. interrogans, MLST and multiple-locus variable-tandem repeat analysis (MLVA) revealed identical ST and MLVA types in rat and canine isolates, whereas two STs and MLVA types were identified in wild boar isolates. The STs and MLVA types of rabbit and one of the wild boars were identical. Bacterial culture and flaB-nested polymerase chain reaction demonstrated a high rate of Leptospira infection in wild boars (58.3%, 7/12), whereas Leptospira spp. were detected in 4.8% of black rats (2/42). This study revealed diverse Leptospira genotype and serotype maintenance in wild mammals on Amami Oshima Island. MLST and MLVA indicated that black rats were a source of canine infection. Wild boars carry L. interrogans and are considered an important maintenance host because antibodies against serogroup Hebdomadis were detected in human and canine leptospirosis patients on this island.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Molecular epidemiology of Leptospira spp. among wild mammals and a dog in Amami Oshima Island, Japan

Shinya¹,

Muraoka

Negishi³

et al. 2021

PLoS ONE

Self Cite

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“…Orthohantaviruses and pathogenic spirochetes of genus Leptospira cause re-emerging global zoonoses clinically indistinguishable in human patients with similar epidemiology [1][2][3][4][5][6]. Rodents are important reservoirs for orthohantavirus and pathogenic leptospira [7].…”

Section: Introductionmentioning

confidence: 99%

Evidence of orthohantavirus and leptospira infections in small mammals in an endemic area of Gampaha district in Sri Lanka

Sunil-Chandra

Fahlman

Waidyarathna

et al. 2022

Preprint

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BackgroundOrthohantaviruses and leptospira are emerging zoonotic pathogens of high public health significance. The epidemiology of orthohantavirus infections and leptospirosis is similar and presents related clinical pictures in humans. However, a paucity of data on actual reservoir hosts for orthohantaviruses and leptospira exists. Therefore, this study aimed at determining the occurrence of orthohantaviruses and leptospira in small mammals captured in an endemic region of Sri Lanka.MethodsRodents and shrews were morphologically and/or genetically identified using morphological keys and DNA barcoding techniques targeting the cytochrome oxidase b subunit gene (Cytb). Lung tissues and sera were subsequently analyzed for the presence of orthohantavirus RNA using qRT-PCR and antibodies against these pathogens.ResultsForty-three (43) small mammals representing: Rattus (R.) rattus (black rat) or R. tanezumi (Asian rat), Suncus murinus (Asian house shrew), R. norvegicus (brown rat) and Mus musculus (house mouse) were investigated. No orthohantavirus RNA was detected from the lung tissue or serum samples of these animals. Elevated levels of IgG antibodies against Puumala orthohantavirus (PUUV) and/or Seoul orthohantavirus (SEOV) antigens were detected in sera of 28 (72%) out of the 39 rats analysed. Interestingly, 36 (92%) of the 39 rats also showed presence of anti leptospira-IgG antibodies in their serum, representing dual infection or dual exposure in 26/39 (66.7%) of examined rats.ConclusionsThis project targets important public health questions concerning the occupational risk of orthohantavirus infections and/or leptospirosis in an endemic region of Sri Lanka. Most rats (72%) in our study displayed antibodies reacting to orthohantavirus NP antigens, related to PUUV and/or SEOV. No correlation between the orthohantavirus and leptospira IgG antibody levels were noticed. Finally, a combination of both morphological and DNA barcoding approaches revealed that several species of rats may play a role in the maintenance and transmission of orthohantavirus and leptospira in Sri Lanka.

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Section: Introductionmentioning

confidence: 99%

Evidence of orthohantavirus and leptospira infections in small mammals in an endemic area of Gampaha district in Sri Lanka

et al. 2022

View full text Add to dashboard Cite

Background Orthohantaviruses and leptospira are emerging zoonotic pathogens of high public health significance. The epidemiology of orthohantavirus infections and leptospirosis is similar and presents related clinical pictures in humans. However, a paucity of data on actual reservoir hosts for orthohantaviruses and leptospira exists. Therefore, this study aimed at determining the occurrence of orthohantaviruses and leptospira in small mammals captured in an endemic region of Sri Lanka. Methods Rodents and shrews were morphologically and/or genetically identified using morphological keys and DNA barcoding techniques targeting the cytochrome oxidase b subunit gene (Cytb). Lung tissues and sera were subsequently analyzed for the presence of orthohantavirus RNA using qRT-PCR. Sera of rats were tested for IgG antibodies against orthohantaviruses and leptospira. Results Forty-three (43) small mammals representing: Rattus (R.) rattus (black rat) or R. tanezumi (Asian rat), Suncus murinus (Asian house shrew), R. norvegicus (brown rat) and Mus musculus (house mouse) were investigated. No orthohantavirus RNA was detected from the lung tissue or serum samples of these animals. Elevated levels of IgG antibodies against Puumala orthohantavirus (PUUV) and/or Seoul orthohantavirus (SEOV) antigens were detected in sera of 28 (72%) out of the 39 rats analysed. Interestingly, 36 (92%) of the 39 rats also showed presence of anti leptospira-IgG antibodies in their serum, representing dual infection or dual exposure in 26/39 (66.7%) of examined rats. Conclusions This project targets important public health questions concerning the occupational risk of orthohantavirus infections and/or leptospirosis in an endemic region of Sri Lanka. Most rats (72%) in our study displayed antibodies reacting to orthohantavirus NP antigens, related to PUUV and/or SEOV. No correlation between the orthohantavirus and leptospira IgG antibody levels were noticed. Finally, a combination of both morphological and DNA barcoding approaches revealed that several species of rats may play a role in the maintenance and transmission of orthohantavirus and leptospira in Sri Lanka.

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Multilocus sequence typing reveals diverse known and novel genotypes of Leptospira spp. circulating in Sri Lanka

Cited by 6 publications

References 32 publications

Molecular epidemiology of Leptospira spp. among wild mammals and a dog in Amami Oshima Island, Japan

Molecular epidemiology of Leptospira spp. among wild mammals and a dog in Amami Oshima Island, Japan

Evidence of orthohantavirus and leptospira infections in small mammals in an endemic area of Gampaha district in Sri Lanka

Evidence of orthohantavirus and leptospira infections in small mammals in an endemic area of Gampaha district in Sri Lanka

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