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Fulltext Leptospira infection and carrier survey on rats from wet market areas in Kuala Lumpur, Malaysia

Hasan T, Afrin S, Sultana A, Islam A

J Vector Borne Dis, 2024 Jan 01;61(1):547-554.
PMID: 38648405 DOI: 10.4103/0972-9062.383644

BACKGROUND OBJECTIVES: Leptospirosis is an important zoonotic infection that has caused significant mortality and morbidity worldwide. This disease is endemic in Malaysia and as a developing tropical country, leptospirosis is concerning as it threatens Malaysian public health and the country's economic sectors. However, there is limited information on leptospirosis in Malaysia, especially regarding leptospiral seroepidemiology among carriers in Malaysia. Therefore, more epidemiological information on the source of the disease and reservoir are needed for better disease control and source intervention. The objectives of this study are to gather information on Leptospira infection and the carrier status of rats captured from selected wet markets of Kuala Lumpur metropolitan city in Malaysia.
METHODS: Live rat trappings were performed in four major wet markets in Kuala Lumpur, namely, Pudu, Chow Kit, Datuk Keramat, and Petaling Street. Animal samplings were performed for 12 months in 2017, where blood and kidney samples were collected and tested for anti-leptospiral antibodies via Microscopic Agglutination Test (MAT) and pathogenic Leptospira screening via Polymerase Chain Reaction (PCR) amplification offlaB gene.
RESULTS: MAT showed that 34.7% (n = 50/144) of the captured rats were positive for anti-leptospiral antibody of which the most prominent serovar was Malaya followed by a local strain, IMR LEP 175. In parallel, 50 rats were also positive for pathogenic Leptospira DNA.
INTERPRETATION CONCLUSION: This study showed that there are persistent Leptospira infections among rats in Kuala Lumpur wet markets and these rats are important reservoir hosts for the bacteria.

Matched MeSH terms: Agglutination Tests
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: Agglutination Tests/standards*
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: Agglutination Tests
Rapid serologic diagnosis of pediatric typhoid fever in an endemic area: a prospective comparative evaluation of two dot-enzyme immunoassays and the Widal test

Bhutta ZA, Mansurali N

Am J Trop Med Hyg, 1999 Oct;61(4):654-7.
PMID: 10548305

We evaluated the diagnostic sensitivity and specificity of two dot-enzyme-linked immunoassays (Typhidot and Typhidot-M; Malaysian Biodiagnostic Research SDN BHD, Kuala Lumpur, Malaysia), assessing IgG and IgM antibodies against the outer membrane protein (OMP) of Salmonella typhi, and the Widal test in comparison with blood culture in a consecutive group of children with suspected typhoid fever. Of 97 children with suspected typhoid fever, the disease was confirmed bacteriologically in 46 (47%), whereas 25 (26%) were considered to have typhoid fever on clinical grounds. An alternative diagnosis was made in 26 (27%). The Typhidot and Typhidot-M were superior to the Widal test in their diagnostic sensitivity and specificity, although values (sensitivity = 85-94% and specificity = 77-89%) were significantly lower than in other regional reports. The lower specificity of the Typhidot in our series may represent regional differences in the genomic structure and plasticity of the OMP of S. typhi and merits further evaluation of these tests in diverse geographic locations.

Matched MeSH terms: Agglutination Tests
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: Agglutination Tests
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: Agglutination Tests
Pathogenic and saprophytic Leptospira species in water and soils from selected urban sites in peninsular Malaysia

Benacer D, Woh PY, Mohd Zain SN, Amran F, Thong KL

Microbes Environ, 2013;28(1):135-40.
PMID: 23363618

Leptospira species were studied in water and soils from selected urban sites in Malaysia. A total of 151 water (n=121) and soil (n=30) samples were collected from 12 recreational lakes and wet markets. All samples were filtered and inoculated into semi-solid Ellinghausen and McCullough modified by Johnson and Harris (EMJH) media supplemented with additional 5-fluorouracil. The cultures were then incubated at 30°C and observed under a dark field microscope with intervals of 10 days. A PCR assay targeting the rrs gene was used to confirm the genus Leptospira among the isolates. Subsequently, the pathogenic status of the isolates was determined using primer sets G1/G2 and Sapro1/Sapro2, which target the secY and rrs genes, respectively. The isolates were identified at serogroup level using the microscopic agglutination test (MAT) while their genetic diversity was assessed by pulsed field gel electrophoresis (PFGE). Based on dark field microscopy, 23.1% (28/121) water and 23.3% (7/30) soil cultures were positive for Leptospira spp. Of the 35 positive cultures, only 8 were pure and confirmed as Leptospira genus by PCR assay. Two out of 8 isolates were confirmed as pathogenic, 5 were saprophytic and one was intermediate. These 8 isolates were negative for the 25 reference hyperimmune rabbit sera tested in the MAT. PFGE showed that all 8 of these environmental Leptospira spp. were genetically diverse. In conclusion, the presence of pathogenic Leptospira spp. in the urban Malaysian environment may indicate and highlight the importance of water screening, especially in recreational lakes, in order to minimize any chance of Leptospira infection.

Matched MeSH terms: Agglutination Tests
Fulltext Molecular characterization of a novel proto-type antimicrobial protein galectin-1 from striped murrel

Arasu A, Kumaresan V, Sathyamoorthi A, Chaurasia MK, Bhatt P, Gnanam AJ, et al.

Microbiol Res, 2014 Nov;169(11):824-34.
PMID: 24780642 DOI: 10.1016/j.micres.2014.03.005

In this study, we reported a molecular characterization of a novel proto-type galectin-1 from the striped murrel Channa striatus (named as CsGal-1). The full length CsGal-1 was identified from an established striped murrel cDNA library and further we confirmed the sequence by cloning. The complete cDNA sequence of CsGal-1 is 590 base pairs (bp) in length and its coding region encoded a poly peptide of 135 amino acids. The polypeptide contains a galactoside binding lectin domain at 4-135. The domain carries a sugar binding site at 45-74 along with its signatures (H(45)-X-Asn(47)-X-Arg(49) and Trp(69)-X-X-Glu(72)-X-Arg(74)). CsGal-1 shares a highly conserved carbohydrate recognition domain (CRD) with galectin-1 from other proto-type galectin of teleosts. The mRNA expressions of CsGal-1 in healthy and various immune stimulants including Aphanomyces invadans, Aeromonas hydrophila, Escherchia coli lipopolysaccharide and poly I:C injected tissues of C. striatus were examined using qRT-PCR. CsGal-1 mRNA is highly expressed in kidney and is up-regulated with different immune stimulants at various time points. To understand its biological activity, the coding region of CsGal-1 gene was expressed in an E. coli BL21 (DE3) cloning system and its recombinant protein was purified. The recombinant CsGal-1 protein was agglutinated with mouse erythrocytes at a concentration of 4μg/mL in a calcium independent manner. CsGal-1 activity was inhibited by d-galactose at 25mM(-1) and d-glucose and d-fructose at 100mM(-1). The results of microbial binding assay showed that the recombinant CsGal-1 protein agglutinated only with the Gram-negative bacteria. Interestingly, we observed no agglutination against Gram-positive bacteria. Overall, the study showed that CsGal-1 is an important immune gene involved in the recognition and elimination of pathogens in C. striatus.

Matched MeSH terms: Agglutination Tests
Serological Prevalence of Leptospirosis Among Rural Communities in the Rejang Basin, Sarawak, Malaysia

Suut L, Mazlan MN, Arif MT, Yusoff H, Abdul Rahim NA, Safii R, et al.

Asia Pac J Public Health, 2016 07;28(5):450-7.
PMID: 27183976 DOI: 10.1177/1010539516648003

Leptospirosis is an important zoonotic disease globally and is endemic in Malaysia. A study was conducted in the Rejang basin of Sarawak from June 2011 to May 2013 to determine the seroprevalence of leptospirosis among the communities and dominant infecting Leptospira serovars. A total of 508 human sera were analyzed using ELISA and the microscopic agglutination test (MAT). The seroprevalence of leptospirosis in the study area was 37.4%, with the highest prevalence in Kapit division. More women were positive for leptospirosis (59.5%), and the mean age of seropositive individuals was 42.2 (SD = 18.7) years. Antibody titers between 1:50 and 1:1600 were reported, and serovars djasiman (22.1%), shermani (13.2%), and pomona (7.9%) predominated, with varied distribution between geographical locations. This study highlighted the endemicity and diversity of existing Leptospira serovars within the community. This information should be communicated to local health personnel and communities at risk, and rapid diagnostic capability should be made available to local health facilities.

Matched MeSH terms: Agglutination Tests
Characterization of group A streptococci isolated in Kuala Lumpur, Malaysia

Jamal F, Pit S, Johnson DR, Kaplan EL

J Trop Med Hyg, 1995 Oct;98(5):343-6.
PMID: 7563264

T-agglutination patterns of 190 strains of group A streptococci isolated between January 1989 and December 1993 from body fluids (10), throat culture (56), pus (51) and skin lesions (73) were determined. Mucoid colonial morphology was exhibited by 6.3% (12/190) of the strains on initial isolation. Type T-5,11,27,44 comprised 23.7%, followed by T-1,3,13,B3264 (11.1%), T-4,6 (8.4%) and T-8,25, Imp 19 (7.9%). About 42% (80/190) strains could not be characterized by T agglutination pattern. T-typing of 71 selected strains at WHO Collaborating Center, Minneapolis yielded similar results. Nineteen selected strains were further characterized by M-typing; only three strains were M-typeable. These strains were isolated from throat (M1), sputum (M5) and pus (M12). About 68% (48/71) isolates produced serum opacity factor. These data support the existence of as yet uncharacterized group A streptococcal serotypes in this region.

Matched MeSH terms: Agglutination Tests
Antigenic potential of a recombinant polyvalent DNA vaccine against pathogenic leptospiral infection

Garba B, Bahaman AR, Zakaria Z, Bejo SK, Mutalib AR, Bande F, et al.

Microb Pathog, 2018 Nov;124:136-144.
PMID: 30138761 DOI: 10.1016/j.micpath.2018.08.028

Leptospirosis is a serious epidemic disease caused by pathogenic Leptospira species. The disease is endemic in most tropical and sub-tropical regions of the world. Currently, there is no effective polyvalent vaccine for prevention against most of the circulating serovars. Moreover, development of an efficient leptospiral vaccine capable of stimulating cross-protective immune responses against a wide range of serovars remains a daunting challenge. This, in part, is associated with the extensive diversity and variation of leptospiral serovars from region to region. In this study, a multi-epitope DNA vaccine encoding highly immunogenic epitopes from LipL32 and LipL41 was designed using in-silico approach. The DNA encoding antigenic epitopes was constructed from conserved pathogenic Leptospira genes (LipL32 and LipL41). Immunization of golden Syrian hamsters with the multi-epitope chimeric DNA vaccine resulted in the production of both agglutinating and neutralizing antibodies as evidence by MAT and in-vitro growth inhibition tests respectively. The antibodies produced reacted against eight different serovars and significantly reduced renal colonization following in vivo challenge. The vaccine was also able to significantly reduce renal colonization which is a very important factor responsible for persistence of leptospires among susceptible and reservoir animal hosts. In conclusion, the leptospiral multi-epitope chimeric DNA vaccine can serve as a potentially effective and safe vaccine against infection with different pathogenic leptospiral serovars.

Matched MeSH terms: Agglutination Tests
An Asian study on the prevalence of atypical respiratory pathogens in community-acquired pneumonia

Ngeow YF, Suwanjutha S, Chantarojanasriri T, Wang F, Saniel M, Alejandria M, et al.

Int J Infect Dis, 2005 May;9(3):144-53.
PMID: 15840455

In many parts of Asia, the inaccessibility and high cost of diagnostic tests have hampered the study of community-acquired pneumonia (CAP) caused by atypical respiratory pathogens.

Matched MeSH terms: Agglutination Tests
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: Agglutination Tests

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