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Leptospirosis in a caver returned from Sarawak, Malaysia

Mortimer RB

Wilderness Environ Med, 2005;16(3):129-31.
PMID: 16209467

This article describes a case of leptospirosis in a man who returned from caving in Sarawak, Malaysia, and includes a discussion of epidemiology, pathophysiology, diagnosis, prevention, and treatment. The patient presented with symptoms of leptospirosis, which was confirmed by microhemagglutination titers. He became infected despite taking doxycycline daily for malaria prophylaxis. Leptospirosis is an important consideration in any returned traveler with fever. The spirochete spreads from animals to humans via water. Caving in tropical endemic zones may increase exposure risk due to the combination of multiple skin abrasions with immersions. Water in caves may increase infection risk because of increased water pH. Standard prophylaxis may be inadequate in cases of high-risk exposures.

Matched MeSH terms: Leptospira/isolation & purification
Fulltext A review of leptospirosis in Malaysia

Bahaman AR, Ibrahim AL

Vet Res Commun, 1988;12(2-3):179-89.
PMID: 3055663 DOI: 10.1007/BF00362799

This paper reviews the literature on leptospirosis in Malaysia from its first description in 1928 until the present day. Most of the early reports were on investigations of leptospirosis in wildlife and man and up-to-date, thirty-seven leptospiral serovars from thirteen serogroups have been bacteriologically identified. The thirteen serogroups are: Australis, Autumnalis Bataviae, Canicola, Celledoni, Grippotyphosa, Hebdomadis, Icterohaemorrhagiae, Javanica, Pomona, Pyrogenes, Sejroe and Tarassovi. Rats have been ascribed as the principal maintenance host of leptospires in Malaysia. However, serovars from the Pomona, Pyrogenes and Sejroe serogroups have yet to be isolated from rats. It is considered that the majority of leptospirosis cases in man were due to association of man with an environment where rats were plentiful. Recent investigations on domestic animals disclosed a high prevalence of infection in cattle and pigs and they were suspected as being the maintenance host for serovar hardjo and pomona respectively. There is ample scope for research in leptospirosis, particularly in the epidemiology and control of the disease in domestic animals. The strategy to control the infection in domestic animals and man in Malaysia is bound to be different from that of the temperate countries, basically due to the presence of a large number of leptospiral serovars in wildlife, further confounded by geographical and financial constraints.

Matched MeSH terms: Leptospira/isolation & purification
Seroprevalence of leptospiral antibodies among market workers and food handlers in the central state of Malaysia

Samsudin S, Sakinah SNS, Malina O, Norliza BA, Noh MA, Fairuz A, et al.

Trop Med Int Health, 2018 03;23(3):327-333.
PMID: 29356240 DOI: 10.1111/tmi.13033

OBJECTIVE: The high prevalence of leptospirosis in humans is of great public health concern, particularly in tropical and subtropical regions. This study aimed to determine the seroprevalence of leptospiral antibodies and distribution of serovars, and to assess the usefulness of enzyme-linked immunosorbent assay (ELISA) as a screening method for leptospiral antibodies in a high-risk healthy community.
METHODS: Cross-sectional study of 231 market workers and food handlers in wet markets and food premises from two localities in central Malaysia. Respondents' background information was obtained using a questionnaire. Serum samples were tested for leptospiral antibodies using ELISA and microscopic agglutination test (MAT).
RESULTS: Seroprevalence of leptospirosis among healthy workers was 46.3%. Detection of seropositivity was higher by MAT (46%) than ELISA (15%). We observed high seropositivity among local workers (49%), food handlers (49.5%), females (60.8%) and those aged 34 years and older (46.3%). Local strain LEP175 was the predominant serovar, followed by WHO strain Patoc.
CONCLUSION: Overall seroprevalence among healthy food handlers and market workers was high in this study. The workplace places susceptible individuals at risk of leptospirosis.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext Rapid detection of pathogenic leptospires by lyophilized reagent-based polymerase chain reaction

Lee SV, Tai ES, Mutalib AR, Khairani-Bejo S, Bahaman AR

Trop Biomed, 2011 Dec;28(3):497-505.
PMID: 22433877 MyJurnal

A simple and reliable tool for the early diagnosis of leptospirosis is urgently needed. We report the development of a lyophilized reagent-based polymerase chain reaction (PCR) assay targeting lipL32 gene, which is present only in pathogenic leptospires. To determine the effectiveness of the newly developed assay in the early diagnosis of leptospirosis, the sensitivity and specificity was evaluated. In simulated clinical samples, the assay was able to detect 10² and 10³ leptospires/ml in spiked urine and blood samples, respectively. In experimentally infected animals, leptospiral DNA could be detected in blood and lung samples as early as Day 1 post infection. This assay was also shown to be stable and remained sensitive for up to five months at ambient temperature. Hence, this lyophilized reagent-based PCR assay with high specificity, sensitivity and stability would provide a simple, rapid and reliable method in diagnosing acute leptospirosis, especially in the field of veterinary medicine.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext Detection of pathogenic Leptospira from selected environment in Kelantan and Terengganu, Malaysia

Ridzlan FR, Bahaman AR, Khairani-Bejo S, Mutalib AR

Trop Biomed, 2010 Dec;27(3):632-8.
PMID: 21399605 MyJurnal

Leptospirosis is recognized as one of the important zoonotic diseases in the world including Malaysia. A total of 145 soil and water samples were collected from selected National Service Training Centres (NSTC) in Kelantan and Terengganu. The samples were inoculated into modified semisolid Ellinghausen McCullough Johnson Harris (EMJH) medium, incubated at room temperature for 1 month and examined under the dark-field microscope. Positive growth of the leptospiral isolates were then confirmed with 8-Azaguanine Test, Polymerase Chain Reaction (PCR) assay and Microscopic Agglutination Test (MAT). Fifteen cultures (10.34%) exhibited positive growths which were seen under dark field microscope whilst only 20% (3/15) were confirmed as pathogenic species. based on 8-Azaguanine Test and PCR. Serological identification of the isolates with MAT showed that hebdomadis was the dominant serovar in Terengganu. Pathogenic leptospires can be detected in Malaysian environment and this has the potential to cause an outbreak. Therefore, precautionary steps against leptospirosis should be taken by camp authorities to ensure the safety of trainees.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext Serological prevalence of leptospiral infection in wild rats at the National Service Training Centres in Kelantan and Terengganu

Mohamed-Hassan SN, Bahaman AR, Mutalib AR, Khairani-Bejo S

Trop Biomed, 2010 Apr;27(1):30-2.
PMID: 20562810 MyJurnal

One hundred and sixty eight rats were trapped from the National Service Training Centres (NSTC) in Kelantan and Terengganu from October 2008 to May 2009. Microscopic agglutination test (MAT) was performed to detect the presence of agglutinating antibodies to Leptospira among the rats caught. All the MAT positive rats were identified as Rattus tiomanicus. In Kelantan, 17.3 % (14/81) of the rats had leptospiral antibodies to serovars Icterohaemorrhagiae (12.3%), Canicola (2.5%), Ballum (1.2%), and Pyrogenes (1.2%). In Terengganu, 18.4% (16/87) of the rats had antibodies to serovars Icterohaemorrhagiae (15%), Canicola (1.1%), Pyrogenes (1.1%) and Hebdomadis (1.1%). This study indicated that Leptospira serovars were prevalent in the rat population in the study areas and could be a source of infection to humans. Therefore, control of the rat population in all NSTC is critical to prevent outbreaks of leptospirosis amongst the NSTC trainees.

Matched MeSH terms: Leptospira/isolation & purification
Fulltext The detection and characterization of pathogenic Leptospira and the use of OMPs as potential antigens and immunogens

Gebriel AM, Subramaniam G, Sekaran SD

Trop Biomed, 2006 Dec;23(2):194-207.
PMID: 17322822 MyJurnal

The detection of leptospires in patient blood in the first week of the disease using PCR provides an early diagnostic tool. PCR using two sets of primers (G1/G2 and B64-I/B64-II) tested with samples seeded with 23 leptospiral strains from pathogenic and non-pathogenic strains was able to amplify leptospiral DNA from pathogenic strains only. Of the 39 antibody negative samples collected from patients suspected for leptospirosis, only 1 sample (2.6%) was PCR positive. Using LSSP-PCR, the G2 primers allowed the characterization of Leptopira species to 10 different genetic signatures which may have epidemiological value in determining species involved in outbreaks. Leptospiral outer membrane proteins from three strains were purified and reacted against patients sera and gave rise to different profiles for pathogenic and non-pathogenic strains. Lymphocytes of mice injected with OMPs proliferated and released IFN(-3) when stimulated in vitro using Leptospira OMP as antigens. This suggests that an immune response could be established using leptospiral OMPs as a putative vaccine. OMPs were also used in a Dot-ELISA to detect antibodies against Leptospira pathogens in humans.

Matched MeSH terms: Leptospira/isolation & purification*
Leptospirosis: An insight into community structure of small mammal's host in urban environment

Mohd-Taib FS, Ishak SN, Yusof MA, Azhari NN, Md-Lasim A, Md Nor S, et al.

Trop Biomed, 2020 Mar 01;37(1):142-154.
PMID: 33612725

Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira and most often acquired through contact with environments contaminated with leptospires shed in the urine of infected mammals. In urban environment, rodents are well-known as the main carriers of this bacteria, however there were no intensive study on the population structure of these animals, and how it associated with this disease. Hence, we use a case study from an outbreak in a residential area in Selangor, Malaysia, to investigate how community structure of small mammals, associated with the prevalence of Leptospira. One hundred cage traps were placed randomly in and around these houses in five phases with two months interval for a year. Community structures (species, sex, and age) were assigned for each individual, prior to screening for pathogenic Leptospira, using a partial lipL32 gene from the kidney samples. 185 small mammals from four species were captured, Rattus norvegicus (74.5%, N=138), R. rattus (20%, N=37), Tupaia glis (5%, N=9), and Suncus murinus (0.5%, N=1). From this number, 29 individuals were found PCR positive for pathogenic Leptospira (R. norvegicus, N=20; R. rattus, N=6; T. glis, N=2; S. murinus, N=1). The study shows that Leptospira occurrence in the small mammals were significantly correlated to age category and sampling phases, with Spearman Correlation (rs) p=0.02 and p=0.04 respectively. Adult individuals were significantly more prevalent with Leptospira infection, whereby March and June were found to associate with higher Leptospira prevalent among the small mammals, potentially coincide with low rainfall and relative humidity level. This information is important in designing a specific control method for rodents in Leptospira outbreak areas. In addition, intensive sampling and regular cleaning effort were found to significantly reduce the small mammal Leptospira reservoir, thus should be implemented in intervention strategies in the urban environment.

Matched MeSH terms: Leptospira/isolation & purification
Serological and bacteriological study of leptospiral infection in a cattle herd in Malaysia

Bahaman AR, Ibrahim AL

Vet Rec, 1986 Sep 27;119(13):325-6.
PMID: 3776042
Matched MeSH terms: Leptospira/isolation & purification
Fulltext Outbreak of melioidosis and leptospirosis co-infection following a rescue operation

Mohamed MS, Khair MT, How SH, Rajalingam R, Sahhir K, Norazah A, et al.

Med J Malaysia, 2012 Jun;67(3):293-7.
PMID: 23082420 MyJurnal

We analyzed the epidemiological data of all people who were involved in the search and rescue operation in Lubuk Yu, a natural recreational forest with waterfall and stream. The hospital admission records of the cases who fulfilled the case definition and the environmental samples result taken at Lubuk Yu recreational area were studied. 153 people were exposed to this outbreak, 85 (55.5%) were professional rescuers from various government agencies and 68 (44.5%) were villagers. 21 fulfilled the case definition. Ten cases were confirmed melioidosis, six melioidosis alone and four coinfected with leptospirosis. There were eight deaths in this outbreak, seven were villagers and one professional rescuer. Overall case fatality was 70%. All confirmed melioidosis cases and seven who died had diabetes mellitus. The morbidity rate were higher among the villagers, 23.5% compared to professional rescuers, 5.9%. The case fatality rate were also higher in this group which was 100% compared to 33.3% in professional rescuers. The soil and water samples in Lubuk Yu recreational area were positive for leptospira and Burkholderia pseudomallei. The presence of co-infection and co-morbidities especially diabetes mellitus among the exposed led to the high mortality in this outbreak hence a high index of suspicion is important among the healthcare professionals in the management of melioidosis cases. To avoid similar incident in future, search and rescue operation should be only conducted by professional rescuers with appropriate personal protective equipment. A register of rescuers should be maintained for surveillance and follow up if necessary.

Matched MeSH terms: Leptospira/isolation & purification
Clinical presentations and predictors of mortality for leptospirosis - A study from suburban area in Malaysia

Fann RJ, Vidya RR, Chong HE, Indralingam V, Christopher Chan WS

Med J Malaysia, 2020 01;75(1):52-56.
PMID: 32008021

INTRODUCTION: Malaysia is endemic for leptospirosis with increasing incidence recorded over the years. Perak has recorded one of the highest incidence and mortality of leptospirosis since 2004.
METHODS: This is a retrospective study of confirmed leptospirosis cases in Larut, Matang and Selama (LMS) district in Perak reported in 2016. The demographic, clinical presentation, laboratory result and clinical outcomes data were analysed and presented.
RESULTS: Forty-two patients with confirmed diagnosis of leptospirosis were included into the study. Majority of patients were males and Malays. The case fatality rate was 14.3%. Patients with leptospirosis present with variable clinical presentations and are commonly seen with coinfection. Patients 70-year-old and older, have clinical presentations suggestive of organ dysfunction and require intensive care are associated with higher mortality.
CONCLUSION: Leptospirosis is endemic in LMS district of Perak with high incidence and case fatality rate. The clinical presentation of leptospirosis is variable. Co-infection of leptospirosis with other acute febrile illness is common. Patients presenting with symptoms and signs of organ dysfunctions or require intensive care are associated with an increased odds of death.

Matched MeSH terms: Leptospira/isolation & purification
Prevalence of leptospiral DNA among wild rodents from a selected area in Beguk Dam Labis, Segamat, Johor, Malaysia

Latifah I, Rahmat MS, Hayarti KB, Paramasvaran S, Azizah MR, Imran F, et al.

Malays J Pathol, 2012 Dec;34(2):157-9.
PMID: 23424779

Leptospirosis is an emerging infectious disease. The differential diagnosis of leptospirosis is difficult due to the varied and often "flu like" symptoms which may result in a missed or delayed diagnosis. Leptospira is the aetiological agent of leptospirosis, a bacterial zoonosis with worldwide distribution. There are over 230 known serovars in the genus Leptospira. The true prevalence of leptospirosis in Malaysia is unknown or underestimated. Our goal was to determine the prevalence for Leptospira infection in rodents in a selected area in Beguk Dam Labis, Segamat, Johor. A study was carried out on 69 serum samples of trapped wild rodents. DNA was extracted from the sera using Leptospira PCR kit (Shanghai ZJ Bio-Tech Co., Ltd). Of 69 rodent serum samples tested by PCR, 9 (13%) showed positive results. In this study we found that (13%) of wild rodents caught in Beguk Dam Labis were infected by Leptospira.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext Fatal co-infection--melioidosis and leptospirosis

Hin HS, Ramalingam R, Chunn KY, Ahmad N, Ab Rahman J, Mohamed MS

Am J Trop Med Hyg, 2012 Oct;87(4):737-40.
PMID: 22826499 DOI: 10.4269/ajtmh.2012.12-0165

Co-infection of melioidosis and leptospirosis is uncommon. We report here four such cases, confirmed by blood culture for melioidosis and blood polymerase-chain reaction for leptospirosis, which occurred among rescuers involved in a search and rescue operation for a young man who was suspected to have drowned in Lubuk Yu, a recreational forest in Pahang, Malaysia. Despite treatment, three of the patients died from the co-infection.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext A duplex endpoint PCR assay for rapid detection and differentiation of Leptospira strains

Benacer D, Zain SNM, Lewis JW, Khalid MKNM, Thong KL

Rev Soc Bras Med Trop, 2017 Mar-Apr;50(2):239-242.
PMID: 28562762 DOI: 10.1590/0037-8682-0364-2016

INTRODUCTION:: This study aimed to develop a duplex endpoint PCR assay for rapid detection and differentiation of Leptospira strains.

METHODS:: Primers were designed to target the rrs (LG1/LG2) and ligB (LP1/LP2) genes to confirm the presence of the Leptospira genus and the pathogenic species, respectively.

RESULTS:: The assay showed 100% specificity against 17 Leptospira strains with a limit of detection of 23.1pg/µl of leptospiral DNA and sensitivity of 103 leptospires/ml in both spiked urine and water.

CONCLUSIONS:: Our duplex endpoint PCR assay is suitable for rapid early detection of Leptospira with high sensitivity and specificity.

Matched MeSH terms: Leptospira/isolation & purification
Fulltext First report of pathogenic Leptospira spp. isolated from urine and kidneys of naturally infected cats

Alashraf AR, Lau SF, Khairani-Bejo S, Khor KH, Ajat M, Radzi R, et al.

PLoS One, 2020;15(3):e0230048.
PMID: 32155209 DOI: 10.1371/journal.pone.0230048

Leptospirosis is one of the most widespread zoonotic diseases and can infect both humans and animals worldwide. Healthy cat, as a potential source of exposure to humans, are likely underestimated owing to the lack of overt clinical signs associated with Leptospira spp. infection in this species. The aim of the study was to determine the exposure, shedding, and carrier status of leptospires in shelter cats in Malaysia by using serological, molecular, and bacteriological methods. For this study, 82 healthy cats from two shelters were sampled. The blood, urine, and kidneys were tested using the microscopic agglutination test (MAT), polymerase chain reaction (PCR), and bacterial culture. On the basis of serological, molecular, and/or culture techniques, the total detection of leptospiral infection was 29.3% (n = 24/82). Through culture techniques, 16.7% (n = 4/24) of the cats that tested positive were carriers with positive kidney cultures, and one cat was culture positive for both urine and kidney. The Leptospira spp. isolates were identified as pathogenic L. interrogans serovar Bataviae through serological and molecular methods. Through serological techniques, 87.5% (n = 21/24) had positive antibody titers (100-1600) and most of the Bataviae serogroup (n = 19/21). Using PCR, 16.7% (n = 4/24) of cats were shown to have pathogenic Leptospira spp. DNA in their urine. Furthermore, three out of four culture positive cats were serology negative. The present study reports the first retrieval of pathogenic leptospires from urine and kidneys obtained from naturally infected cats. The results provide evidence of the potential role of naturally infected cats in the transmission of leptospires. Additionally, leptospiral infection occurs sub-clinically in cats. The culture isolation provides evidence that healthy cats could be reservoirs of leptospiral infection, and this information may promote the development of disease prevention strategies for the cat population.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext Leptospira infection in rats: A literature review of global prevalence and distribution

Boey K, Shiokawa K, Rajeev S

PLoS Negl Trop Dis, 2019 08;13(8):e0007499.
PMID: 31398190 DOI: 10.1371/journal.pntd.0007499

BACKGROUND: The role of rodents in Leptospira epidemiology and transmission is well known worldwide. Rats are known to carry different pathogenic serovars of Leptospira spp. capable of causing disease in humans and animals. Wild rats (Rattus spp.), especially the Norway/brown rat (Rattus norvegicus) and the black rat (R. rattus), are the most important sources of Leptospira infection, as they are abundant in urban and peridomestic environments. In this study, we compiled and summarized available data in the literature on global prevalence of Leptospira exposure and infection in rats, as well as compared the global distribution of Leptospira spp. in rats with respect to prevalence, geographic location, method of detection, diversity of serogroups/serovars, and species of rat.
METHODS: We conducted a thorough literature search using PubMed without restrictions on publication date as well as Google Scholar to manually search for other relevant articles. Abstracts were included if they described data pertaining to Leptospira spp. in rats (Rattus spp.) from any geographic region around the world, including reviews. The data extracted from the articles selected included the author(s), year of publication, geographic location, method(s) of detection used, species of rat(s), sample size, prevalence of Leptospira spp. (overall and within each rat species), and information on species, serogroups, and/or serovars of Leptospira spp. detected.
FINDINGS: A thorough search on PubMed retrieved 303 titles. After screening the articles for duplicates and inclusion/exclusion criteria, as well as manual inclusion of relevant articles, 145 articles were included in this review. Leptospira prevalence in rats varied considerably based on geographic location, with some reporting zero prevalence in countries such as Madagascar, Tanzania, and the Faroe Islands, and others reporting as high as >80% prevalence in studies done in Brazil, India, and the Philippines. The top five countries that were reported based on number of articles include India (n = 13), Malaysia (n = 9), Brazil (n = 8), Thailand (n = 7), and France (n = 6). Methods of detecting or isolating Leptospira spp. also varied among studies. Studies among different Rattus species reported a higher Leptospira prevalence in R. norvegicus. The serovar Icterohaemorrhagiae was the most prevalent serovar reported in Rattus spp. worldwide. Additionally, this literature review provided evidence for Leptospira infection in laboratory rodent colonies within controlled environments, implicating the zoonotic potential to laboratory animal caretakers.
CONCLUSIONS: Reports on global distribution of Leptospira infection in rats varies widely, with considerably high prevalence reported in many countries. This literature review emphasizes the need for enhanced surveillance programs using standardized methods for assessing Leptospira exposure or infection in rats. This review also demonstrated several weaknesses to the current methods of reporting the prevalence of Leptospira spp. in rats worldwide. As such, this necessitates a call for standardized protocols for the testing and reporting of such studies, especially pertaining to the diagnostic methods used. A deeper understanding of the ecology and epidemiology of Leptospira spp. in rats in urban environments is warranted. It is also pertinent for rat control programs to be proposed in conjunction with increased efforts for public awareness and education regarding leptospirosis transmission and prevention.

Matched MeSH terms: Leptospira/isolation & purification
Fulltext Emerging Infectious Disease Implications of Invasive Mammalian Species: The Greater White-Toothed Shrew (Crocidura russula) Is Associated With a Novel Serovar of Pathogenic Leptospira in Ireland

Nally JE, Arent Z, Bayles DO, Hornsby RL, Gilmore C, Regan S, et al.

PLoS Negl Trop Dis, 2016 12;10(12):e0005174.
PMID: 27935961 DOI: 10.1371/journal.pntd.0005174

The greater white-toothed shrew (Crocidura russula) is an invasive mammalian species that was first recorded in Ireland in 2007. It currently occupies an area of approximately 7,600 km2 on the island. C. russula is normally distributed in Northern Africa and Western Europe, and was previously absent from the British Isles. Whilst invasive species can have dramatic and rapid impacts on faunal and floral communities, they may also be carriers of pathogens facilitating disease transmission in potentially naive populations. Pathogenic leptospires are endemic in Ireland and a significant cause of human and animal disease. From 18 trapped C. russula, 3 isolates of Leptospira were cultured. However, typing of these isolates by standard serological reference methods was negative, and suggested an, as yet, unidentified serovar. Sequence analysis of 16S ribosomal RNA and secY indicated that these novel isolates belong to Leptospira alstonii, a unique pathogenic species of which only 7 isolates have been described to date. Earlier isolations were limited geographically to China, Japan and Malaysia, and this leptospiral species had not previously been cultured from mammals. Restriction enzyme analysis (REA) further confirms the novelty of these strains since no similar patterns were observed with a reference database of leptospires. As with other pathogenic Leptospira species, these isolates contain lipL32 and do not grow in the presence of 8-azagunaine; however no evidence of disease was apparent after experimental infection of hamsters. These isolates are genetically related to L. alstonii but have a novel REA pattern; they represent a new serovar which we designate as serovar Room22. This study demonstrates that invasive mammalian species act as bridge vectors of novel zoonotic pathogens such as Leptospira.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext Revisiting the taxonomy and evolution of pathogenicity of the genus Leptospira through the prism of genomics

Vincent AT, Schiettekatte O, Goarant C, Neela VK, Bernet E, Thibeaux R, et al.

PLoS Negl Trop Dis, 2019 05;13(5):e0007270.
PMID: 31120895 DOI: 10.1371/journal.pntd.0007270

The causative agents of leptospirosis are responsible for an emerging zoonotic disease worldwide. One of the major routes of transmission for leptospirosis is the natural environment contaminated with the urine of a wide range of reservoir animals. Soils and surface waters also host a high diversity of non-pathogenic Leptospira and species for which the virulence status is not clearly established. The genus Leptospira is currently divided into 35 species classified into three phylogenetic clusters, which supposedly correlate with the virulence of the bacteria. In this study, a total of 90 Leptospira strains isolated from different environments worldwide including Japan, Malaysia, New Caledonia, Algeria, mainland France, and the island of Mayotte in the Indian Ocean were sequenced. A comparison of average nucleotide identity (ANI) values of genomes of the 90 isolates and representative genomes of known species revealed 30 new Leptospira species. These data also supported the existence of two clades and 4 subclades. To avoid classification that strongly implies assumption on the virulence status of the lineages, we called them P1, P2, S1, S2. One of these subclades has not yet been described and is composed of Leptospira idonii and 4 novel species that are phylogenetically related to the saprophytes. We then investigated genome diversity and evolutionary relationships among members of the genus Leptospira by studying the pangenome and core gene sets. Our data enable the identification of genome features, genes and domains that are important for each subclade, thereby laying the foundation for refining the classification of this complex bacterial genus. We also shed light on atypical genomic features of a group of species that includes the species often associated with human infection, suggesting a specific and ongoing evolution of this group of species that will require more attention. In conclusion, we have uncovered a massive species diversity and revealed a novel subclade in environmental samples collected worldwide and we have redefined the classification of species in the genus. The implication of several new potentially infectious Leptospira species for human and animal health remains to be determined but our data also provide new insights into the emergence of virulence in the pathogenic species.

Matched MeSH terms: Leptospira/isolation & purification
Fulltext Diagnostic accuracy of genetic markers and nucleic acid techniques for the detection of Leptospira in clinical samples: A meta-analysis

Lam JY, Low GK, Chee HY

PLoS Negl Trop Dis, 2020 02;14(2):e0008074.
PMID: 32049960 DOI: 10.1371/journal.pntd.0008074

BACKGROUND: Leptospirosis is often difficult to diagnose because of its nonspecific symptoms. The drawbacks of direct isolation and serological tests have led to the increased development of nucleic acid-based assays, which are more rapid and accurate. A meta-analysis was performed to evaluate the diagnostic accuracy of genetic markers for the detection of Leptospira in clinical samples.
METHODOLOGY AND PRINCIPLE FINDINGS: A literature search was performed in Scopus, PubMed, MEDLINE and non-indexed citations (via Ovid) by using suitable keyword combinations. Studies evaluating the performance of nucleic acid assays targeting leptospire genes in human or animal clinical samples against a reference test were included. Of the 1645 articles identified, 42 eligible studies involving 7414 samples were included in the analysis. The diagnostic performance of nucleic acid assays targeting the rrs, lipL32, secY and flaB genes was pooled and analyzed. Among the genetic markers analyzed, the secY gene showed the highest diagnostic accuracy measures, with a pooled sensitivity of 0.56 (95% CI: 0.50-0.63), a specificity of 0.98 (95% CI: 0.97-0.98), a diagnostic odds ratio of 46.16 (95% CI: 6.20-343.49), and an area under the curve of summary receiver operating characteristics curves of 0.94. Nevertheless, a high degree of heterogeneity was observed in this meta-analysis. Therefore, the present findings here should be interpreted with caution.
CONCLUSION: The diagnostic accuracies of the studies examined for each genetic marker showed a significant heterogeneity. The secY gene exhibited higher diagnostic accuracy measures compared with other genetic markers, such as lipL32, flaB, and rrs, but the difference was not significant. Thus, these genetic markers had no significant difference in diagnostic accuracy for leptospirosis. Further research into these genetic markers is warranted.

Matched MeSH terms: Leptospira/isolation & purification*
Fulltext Leptospira interrogans and Leptospira kirschneri are the dominant Leptospira species causing human leptospirosis in Central Malaysia

Philip N, Bahtiar Affendy N, Ramli SNA, Arif M, Raja P, Nagandran E, et al.

PLoS Negl Trop Dis, 2020 Mar;14(3):e0008197.
PMID: 32203511 DOI: 10.1371/journal.pntd.0008197

BACKGROUND: Leptospirosis, commonly known as rat-urine disease, is a global but endemic zoonotic disease in the tropics. Despite the historical report of leptospirosis in Malaysia, the information on human-infecting species is limited. Determining the circulating species is important to understand its epidemiology, thereby to strategize appropriate control measures through public health interventions, diagnostics, therapeutics and vaccine development.
METHODOLOGY/PRINCIPLE FINDINGS: We investigated the human-infecting Leptospira species in blood and serum samples collected from clinically suspected leptospirosis patients admitted to three tertiary care hospitals in Malaysia. From a total of 165 patients, 92 (56%) were confirmed cases of leptospirosis through Microscopic Agglutination Test (MAT) (n = 43; 47%), Polymerase Chain Reaction (PCR) (n = 63; 68%) or both MAT and PCR (n = 14; 15%). The infecting Leptospira spp., determined by partial 16S rDNA (rrs) gene sequencing revealed two pathogenic species namely Leptospira interrogans (n = 44, 70%) and Leptospira kirschneri (n = 17, 27%) and one intermediate species Leptospira wolffii (n = 2, 3%). Multilocus sequence typing (MLST) identified an isolate of L. interrogans as a novel sequence type (ST 265), suggesting that this human-infecting strain has a unique genetic profile different from similar species isolated from rodents so far.
CONCLUSIONS/SIGNIFICANCE: Leptospira interrogans and Leptospira kirschneri were identified as the dominant Leptospira species causing human leptospirosis in Central Malaysia. The existence of novel clinically important ST 265 (infecting human), that is different from rodent L. interrogans strains cautions reservoir(s) of these Leptospira lineages are yet to be identified.

Matched MeSH terms: Leptospira/isolation & purification*

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