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Fulltext Case Report: Fatal Human Leptospirosis Caused by Leptospira interrogans Genotype ST149

Rao M, Amran F, Kamaruzaman AA, Hakim Esa HA, Abdul Hameed A, Mohamed Shabery NA

Am J Trop Med Hyg, 2021 Jan;104(1):216-218.
PMID: 33289472 DOI: 10.4269/ajtmh.20-0267

Leptospirosis is an important zoonotic disease with worldwide distribution and nonspecific clinical manifestation. We report a case of fatal leptospirosis in a previously healthy woman with a causative agent. A young adult Indian woman was brought in dead to the forensic department. Ten days before, she developed fever, dizziness with headache, myalgia, diarrhea, and vomiting. Routine inquest and autopsy were performed on the deceased, revealing hemorrhagic lungs with extensive intra-alveolar hemorrhages, pale liver with dissociation and separation of hepatocyte plates, and edematous brain with histiocyte and lymphocyte infiltration in the parenchyma and meninges. Heart tissue depicts myocarditis and pericarditis inflammatory changes. Cerebrospinal fluid (CSF) was turbid in appearance with mildly elevated leukocytes, predominantly lymphocytes. Real-time PCR targeting lipL32 gene of pathogenic Leptospira was detected in the blood, CSF, brain, kidney, heart, and liver. The genetic profile of the causative agent was ST149 (multi-locus sequence typing Scheme 3). This study illustrates the usefulness of Leptospira PCR assay in postmortem diagnosis and addresses the need for further surveillance to identify the epidemiological link of the disease.

Matched MeSH terms: Leptospira interrogans/classification; Leptospira interrogans/genetics*
Fulltext Isolation and molecular characterization of Leptospira interrogans and Leptospira borgpetersenii isolates from the urban rat populations of Kuala Lumpur, Malaysia

Benacer D, Mohd Zain SN, Amran F, Galloway RL, Thong KL

Am J Trop Med Hyg, 2013 Apr;88(4):704-9.
PMID: 23358635 DOI: 10.4269/ajtmh.12-0662

Rats are considered the principal maintenance hosts of Leptospira. The objectives of this study were isolation and identification of Leptospira serovars circulating among urban rat populations in Kuala Lumpur. Three hundred urban rats (73% Rattus rattus and 27% R. norvegicus) from three different sites were trapped. Twenty cultures were positive for Leptospira using dark-field microscopy. R. rattus was the dominant carrier (70%). Polymerase chain reaction (PCR) confirmed that all isolates were pathogenic Leptospira species. Two Leptospira serogroups, Javanica and Bataviae, were identified using microscopic agglutination test (MAT). Pulsed-field gel electrophoresis (PFGE) identified two serovars in the urban rat populations: L. borgpetersenii serovar Javanica (85%) and L. interrogans serovar Bataviae (15%). We conclude that these two serovars are the major serovars circulating among the urban rat populations in Kuala Lumpur. Despite the low infection rate reported, the high pathogenicity of these serovars raises concern of public health risks caused by rodent transmission of leptospirosis.

Matched MeSH terms: Leptospira interrogans/classification; Leptospira interrogans/genetics; Leptospira interrogans/isolation & purification*
Draft Genome Sequence of Leptospira interrogans Serovar Bataviae Strain LepIMR 22 Isolated from a Rodent in Johor, Malaysia

Amran F, Mohd Khalid MK, Mohamad S, Mat Ripen A, Ahmad N, Goris MG, et al.

Genome Announc, 2016;4(5).
PMID: 27609924 DOI: 10.1128/genomeA.00956-16

Leptospira interrogans serovar Bataviae was recently identified as one of the persistent Leptospira serovars in Malaysia. Here, we report the draft genome sequence of the L. interrogans serovar Bataviae strain LepIMR 22 isolated from kidney of a rodent in Johor, Malaysia.

Matched MeSH terms: Leptospira interrogans
Role of 72 kDa protein of Leptospira interrogans as a diagnostic marker in acute leptospirosis

Riazi M, Zainul FZ, Bahaman AR, Amran F, Khalilpour A

Indian J Med Res, 2014 Feb;139(2):308-13.
PMID: 24718408

Leptospirosis is a widespread zoonotic disease and a public health problem, particularly in tropical and subtropical countries. Varied clinical manifestations of the disease frequently lead to misdiagnosis resulting in life-threatening multi-organ complications. Therefore, early laboratory investigation using an appropriate diagnostic approach is crucial. In the present study, a potential protein marker was identified and evaluated for its usefulness in the serodiagnosis of acute leptospirosis.

Matched MeSH terms: Leptospira interrogans serovar icterohaemorrhagiae/genetics*; Leptospira interrogans serovar icterohaemorrhagiae/immunology; Leptospira interrogans serovar icterohaemorrhagiae/pathogenicity
rRNA gene restriction patterns of Leptospira: a molecular typing system

Pérolat P, Grimont F, Regnault B, Grimont PA, Fournié E, Thevenet H, et al.

Res. Microbiol., 1990 Feb;141(2):159-71.
PMID: 2189169

A total of 67 serovar reference strains and 7 isolates belonging to the genus Leptospira were characterized by ribosomal ribonucleic acid (rRNA) gene restriction patterns. Fifty patterns were observed. Strains belonging to different genomic species always gave different patterns. However, genomic species were subdivided into several patterns. Forty-three serovars gave a specific pattern. Some serovars could not be separated by rRNA gene restriction patterns: strains of serovars icterohaemorrhagiae, copenhageni, lai, pyrogenes and jalna gave pattern 1; serovars birkini, mankarso and wolffi gave pattern 4; serovars canicola, gem, hebdomadis, pomona and hardjo (strain hardjoprajitno) gave pattern 12; serovars valbuzzi and zanoni gave pattern 14; serovars jonsis, malaya and sumneri gave pattern 16; serovars arborea, ballum, castellonis and kenya gave pattern 35; and serovars borincana and shermani gave pattern 43. These data provide the bases for a molecular typing system for the genus Leptospira.

Matched MeSH terms: Leptospira interrogans/classification; Leptospira interrogans/genetics
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 interrogans serovar canicola/immunology; Leptospira interrogans serovar canicola/isolation & purification
The bacteriological prevalence of leptospiral infection in cattle and buffaloes in West Malaysia

Bahaman AR, Ibrahim AL, Stallman ND, Tinniswood RD

Epidemiol Infect, 1988 Apr;100(2):239-46.
PMID: 3356222

A cross-sectional bacteriological survey of cattle in West Malaysia revealed 14.4% (32/222) had leptospiral infection. Isolates were obtained from all except one herd with prevalence of infection in herds ranging from 0-44.8%. A small number of buffalo urine samples were examined and all of them were found to be negative. A leptospiral isolate obtained from a bovine kidney proved to be a new serovar of Leptospira interrogans and the name unipertama was assigned to it. Six other leptospiral serovars were isolated, namely canicola, australis, javanica, ballum, pomona and hardjo. All six serovars were isolated for the first time in cattle in Malaysia. Cattle in Malaysia appear to be the maintenance host for serovar hardjo. The presence of the other serovars in cattle was probably due to contact with the maintenance hosts, pigs for serovar pomona and rodents for the other three serovars. It appears that the epidemiology of leptospiral infection in cattle in Malaysia is similar to that reported overseas.

Matched MeSH terms: Leptospira interrogans/isolation & purification; Leptospira interrogans serovar canicola/isolation & purification
Fulltext Seroprevalence of Leptospira infection in occupational risk groups in North Khorasan province, Iran

Hashemi SA, Arzamani K, Abdollahpour G, Beheshti N, Alavinia M, Azimian A, et al.

Heliyon, 2021 Jan;7(1):e05983.
PMID: 33506135 DOI: 10.1016/j.heliyon.2021.e05983

Leptospirosis is an important zoonotic bacterial disease caused by Leptospira spp. Earlier studies from North Khorasan province (Iran) reported the presence of Leptospira in wild canines and rodents. To date, there is no data on the seroprevalence of leptospirosis among humans in this province. This study was performed to determine the prevalence of human leptospiral infection among people with different occupations. The study was conducted in urban and rural areas of the province. Among the serum samples collected from 278 subjects, 3 (1.1%) showed positive reaction with titer of 1:100 by the microscopic agglutination test (MAT). Positive reactions were detected against Leptospira interrogans Canicola and L. interrogans icterohemorrhagic and all these samples were from livestock farmers (n = 3/106, 2.7%). The current study revealed that, though Leptospira infection is low in North Khorasan province, regular monitoring of the livestock and the farmers are important.

Matched MeSH terms: Leptospira interrogans
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 interrogans/genetics; Leptospira interrogans/isolation & purification*; Leptospira interrogans/pathogenicity
Serological prevalence of leptospiral infection in domestic animals in West Malaysia

Bahaman AR, Ibrahim AL, Adam H

Epidemiol Infect, 1987 Oct;99(2):379-92.
PMID: 3678399

A cross-sectional serological survey of domestic animals in West Malaysia revealed that 25.5% of the animals examined had agglutinating antibodies to one or more antigens belonging to Leptospira interrogans. Significant prevalence of infection was observed in cattle (40.5%), buffaloes (31%) and pigs (16%). The Sejroe serogroup was shown to be the principal one involved in cattle and buffaloes, and to a lesser extent the Tarassovi and Pomona serogroups. Evidence of infection in domestic animals by strains bearing the other seven antigens appeared insignificant and was indicative of sporadic infection. A majority of the large (semi-intensive) cattle and buffalo farms demonstrated a high prevalence of leptospiral infection. In both species of domestic animals mentioned above, the prevalence of infection was significantly higher (P = 0.01) in the semi-intensive farms than in the smallholdings. Amongst cattle, the droughtmasters had the highest prevalence whilst the Kedah-Kelantan (an indigenous breed) had the lowest prevalence of leptospiral infection. In general, the temperate breeds of cattle had a significantly (P = 0.01) higher prevalence of infection than local breeds. Leptospiral infection in goats and sheep was shown to be sporadic, and the Pomona serogroup was the principal leptospiral serogroup involved in these small ruminants. The prevalence of infection in pigs was observed to decline during the study period, and it is suspected that pigs in West Malaysia are the maintenance host for serovar pomona whilst cattle are the maintenance host for serovar hardjo. Overall, it appears that domestic animals in Malaysia will play a bigger role in the epidemiology of leptospiral infection with the advent of sophisticated farming.

Matched MeSH terms: Leptospira interrogans/immunology
Fulltext A low molecular weight lipopolysaccharide antigen preparation reactive to acute leptospirosis heterologous sera

Riazi M, Abdul Rani B, Fairuz A, Zainul FZ

Trop Biomed, 2010 Aug;27(2):241-53.
PMID: 20962722 MyJurnal

There is a need for identification of new infection markers against common Leptospira isolates in Malaysia. To achieve this goal, seven-day-old cultures of Leptospira interrogans serogroup Icterohemorrhagiae (L44) and Leptospira interrogans serogroup Javanica (L55) were used for antigen preparation by sequential extraction method using 40 mM Tris, 8M Urea and 2M thiourea. Immunoblot analysis of the antigens were performed using serum samples from 46 local patients with confirmed acute leptospirosis, 28 patients with other infections and 14 healthy controls. The patients serum samples used in this study contained heterologous antibody against a number of different leptospira serovars. A strong IgM reactivity to a broad diffuse band of 10-15 kDa was observed. Combining results using L44 and L55 antigens showed sensitivity of 80.4% and specificity of 95.2% for detection of leptospirosis. Proteinase K and periodate treatment indicated that the band is likely to be lipopolysaccharide (LPS) in nature. This study showed that the 10-15 kDa antigen could potentially be useful for serodiagnosis of acute leptospirosis in Malaysia.

Matched MeSH terms: Leptospira interrogans/immunology*
Fulltext Vascular pathology associated with Leptospira icterohaemorrhagiae serovar Lai strain Langkawi infection in guinea pigs (Cavia porcellus)

Bahaman, A.R., Fuzina, N.H., Tengku-Azmi, T.I., Tyagita, G.H., Jasni, S.

Jurnal Veterinar Malaysia, 2018;30(1):7-14.
MyJurnal

Fifteen (15) guinea pigs were experimentally infected with Leptospira icterohemorrhagiae serovar Lai strain
Langkawi, a new strain that was isolated from a human leptospirosis patient. Hematoxylin and Eosin ((H&E) staining
showed haemorrhages, congestion and oedema in all internal organs examined (lungs, liver, spleen and kidneys) with
inflammatory cell infiltration characterized by neutrophils, lymphocytes and macrophages. Hydropic degeneration and
cell necrosis were also common in our findings. Leptospires were detected starting Day 2 p.i by silver staining and
Transmission Electron Microscopy (TEM). Rise in antibody titres started on Day 5 p.i and leptospiral DNA was
detected beginning Day 3 in the kidneys and Day 5 in the liver by Polymerase Chain Reaction (PCR) assay. The
findings illustrated the pathogenesis of leptospirosis in guinea pigs which disclosed them as a suitable animal model for
demonstration of clinical symptoms of leptospirosis and pathological changes after being infected with Leptospira
icterohaemorrhagiae serovar Lai strain Langkawi, particularly pulmonary haemorrhages, a leading cause of mortality
in human leptospirosis.

Matched MeSH terms: Leptospira interrogans serovar icterohaemorrhagiae
Development of monoclonal antibodies against recombinant LipL21 protein of pathogenic Leptospira through phage display technology

Mohd Ali MR, Sum JS, Aminuddin Baki NN, Choong YS, Nor Amdan NA, Amran F, et al.

Int J Biol Macromol, 2021 Jan 31;168:289-300.
PMID: 33310091 DOI: 10.1016/j.ijbiomac.2020.12.062

Leptospirosis is a potentially fatal zoonosis that is caused by spirochete Leptospira. The signs and symptoms of leptospirosis are usually varied, allowing it to be mistaken for other causes of acute febrile syndromes. Thus, early diagnosis and identification of a specific agent in clinical samples is crucial for effective treatment. This study was aimed to develop specific monoclonal antibodies against LipL21 antigen for future use in leptospirosis rapid and accurate immunoassay. A recombinant LipL21 (rLipL21) antigen was optimized for expression and evaluated for immunogenicity. Then, a naïve phage antibody library was utilized to identify single chain fragment variable (scFv) clones against the rLipL21 antigen. A total of 47 clones were analysed through monoclonal phage ELISA. However, after taking into consideration the background OD405 values, only 4 clones were sent for sequencing to determine human germline sequences. The sequence analysis showed that all 4 clones are identical. The in silico analysis of scFv-lip-1 complex indicated that the charged residues of scFv CDRs are responsible for the recognition with rLipL21 epitopes. The generated monoclonal antibody against rLipL21 will be evaluated as a detection reagent for the diagnosis of human leptospirosis in a future study.

Matched MeSH terms: Leptospira interrogans/genetics
Fulltext Leptospirosis in Kuala Lumpur and the comparative evaluation of two rapid commercial diagnostic kits against the MAT test for the detection of antibodies to leptospira interrogans

Sekhar WY, Soo EH, Gopalakrishnan V, Devi S

Singapore Med J, 2000 Aug;41(8):370-5.
PMID: 11256343

The aim of the study was to look into the epidemiology of serodiagnosed cases of leptospirosis at the University Hospital and compare two commercial ELISA Assays to the Microscopic Agglutination Test (MAT). Demographic data for all serodiagnosed cases for the years 1991-1997 were collected. From this data, 104 sera (n = 104) were selected as samples for comparative evaluation of the commercial ELISAs (INDX Dip-S-Ticks and PanBio ELISA) to the MAT test. Thirty two (n = 32) negative control sera were selected from serodiagnosed cases of other differential diagnosis of leptospira infection. The MAT test is a standard test that detects agglutination antibodies to leptospira biflexa, while the INDX Dip-S-Ticks is an ELISA dot test assaying for total anti-leptospira antibodies. The PanBio ELISA is a colorometric assay in test well strips to detect anti-leptospira IgM. The sensitivity, specificity, and efficiency of tests were calculated at a MAT cut-off value of 1:320. Demographic data showed that leptospirosis peaks during March-May and Aug-Nov coinciding with the inter-monsoon period with more men being infected than women and more adults than children. The sensitivity, specificity, and efficiency of test for the INDX Dip-S-Ticks were 83.3%, 93.8% and 87.5% while the values for the PanBio ELISA were 54.2%, 96.9% and 71.3%. The suboptimal PanBio result could be related to the blocking effect of high IgG titres or could be related to the diagnostic MAT cut-off values used in this study. The data hence reflects a pattern of transmission that is related to "wet" occupational risk factors. The commercial assays evaluated, are easier to perform but interpretation of results should be based on level of endemicity. The INDX Dip-S-Ticks allows this flexibility and is a practical alternative to the MAT test.

Matched MeSH terms: Leptospira interrogans/immunology*

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