Non-linear effect of different humidity types on scrub typhus occurrence in endemic provinces, Thailand.

BACKGROUND: Reported monthly scrub typhus (ST) cases in Thailand has an increase in the number of cases during 2009-2014. Humidity is a crucial climatic factor for the survival of chiggers, which is the disease vectors. The present study was to determine the role of humidity in ST occurrence in Thailand and its delayed effect. METHODS: We obtained the climate data from the Department of Meteorology, the disease data from Ministry of Public Health. Negative binomial regression combined with a distributed lag non-linear model (NB-DLNM) was employed to determine the non-linear effects of different types of humidity on the disease. This model controlled overdispersion and confounder, including seasonality, minimum temperature, and cumulative total rainwater. RESULTS: The occurrence of the disease in the 6-year period showed the number of cases gradually increased summer season (Mid-February - Mid-May) and then reached a plateau during the rainy season (Mid-May - Mid-October) and then steep fall after the cold season (Mid-October - Mid-February). The high level (at 70%) of minimum relative humidity (RHmin) was associated with a 33% (RR 1.33, 95% CI 1.13-1.57) significant increase in the number of the disease; a high level (at 14 g/m3) of minimum absolute humidity (AHmin) was associated with a 30% (RR 1.30, 95% CI 1.14-1.48); a high level (at 1.4 g/kg) of minimum specific humidity (SHmin) was associated with a 28% (RR 1.28, 95% CI 1.04-1.57). The significant effects of these types of humidity occurred within the past month. CONCLUSION: Humidity played a significant role in enhancing ST cases in Thailand, particularly at a high level and usually occurred within the past month. NB-DLNM had good controlled for the overdispersion and provided the precise estimated relative risk of non-linear associations. Results from this study contributed the evidence to support the Ministry of Public Health on warning system which might be useful for public health intervention and preparation in Thailand.

Human single-chain variable fragment antibody expressed in E. coli with optimal in vitro cross-neutralizing and no enhancing activity.

Single chain fragment variable (scFv) is a small molecule antibody comprising of only the variable region of heavy and light chain responsible for antigen binding. For dengue disease, the Fc region of antibody molecule was reported to be involved with dengue complication caused by Antibody-dependent enhancement (ADE). We attempted to produce small molecule scFv human monoclonal antibody (HuMAb), which lacking the Fc portion to eliminate the ADE effect of the IgG. This scFv antibody was produced in Escherichia coli. The biologically active form of scFv antibody was successfully generated. 23-1C2D2-scFv showed neutralizing activity similar to the IgG obtained from parental hybridoma, but lacked enhancing activity in all studied concentrations. This antibody was targeted to the 101WXN103 motif of dengue envelop protein domain II, studied by western blot analysis with truncated E protein and random peptide phage display. This scFv is verified as a candidate for further development as therapeutic candidate for DENV infection.

Generation and characterization of cross neutralizing human monoclonal antibody against 4 serotypes of dengue virus without enhancing activity.

BACKGROUND: Dengue disease is a leading cause of illness and death in the tropics and subtropics. Most severe cases occur among patients secondarily infected with a different dengue virus (DENV) serotype compared with that from the first infection, resulting in antibody-dependent enhancement activity (ADE). Our previous study generated the neutralizing human monoclonal antibody, D23-1B3B9 (B3B9), targeting the first domain II of E protein, which showed strong neutralizing activity (NT) against all four DENV serotypes. However, at sub-neutralizing concentrations, it showed ADE activity in vitro. METHODS: In this study, we constructed a new expression plasmid using the existing IgG heavy chain plasmid as a template for Fc modification at position N297Q by site-directed mutagenesis. The resulting plasmid was then co-transfected with a light chain plasmid to produce full recombinant IgG (rIgG) in mammalian cells (N297Q-B3B9). This rIgG was characterized for neutralizing and enhancing activity by using different FcγR bearing cells. To produce sufficient quantities of B3B9 rIgG for further characterization, CHO-K1 cells stably secreting N297Q-B3B9 rIgG were then established. RESULTS: The generated N297Q-B3B9 rIgG which targets the conserved N-terminal fusion loop of DENV envelope protein showed the same cross-neutralizing activity to all four DENV serotypes as those of wild type rIgG. In both FcγRI- and RII-bearing THP-1 cells and FcγRII-bearing K562 cells, N297Q-B3B9 rIgG lacked ADE activity against all DENV serotypes at sub-neutralizing concentrations. Fortunately, the N297Q-B3B9 rIgG secreted from stable cells showed the same patterns of NT and ADE activities as those of the N297Q-B3B9 rIgG obtained from transient expression against DENV2. Thus, the CHO-K1 stably expressing N297Q-B3B9 HuMAb can be developed as high producer stable cells and used to produce sufficient amounts of antibody for further characterization as a promising dengue therapeutic candidate. DISCUSSION: Human monoclonal antibody, targeted to fusion loop of envelope domainII (EDII), was generated and showed cross-neutralizing activity to 4 serotypes of DENV, but did not cause any viral enhancement activity in vitro. This HuMAb could be further developed as therapeutic candidates.

Migration, health, and socioenvironmental safety net among children of Dhaka, Bangladesh.

This study quantifies the diarrhea burden among migrant children under age 5 (who have migrated due to environmental degradation) in Dhaka. We used a multifactor socioepidemiological as well as environmental approach with pretested questionnaires and observations. It was found that 52% of the children were affected by diarrhea. Disability-adjusted life years (DALYs) lost was reduced manifold with the increase of mothers' behavioral determinants. Health losses were 1,718 fold with significant coefficient (ß) in the migrant group. DALYs lost were significantly associated with socioenvironmental factors such as mother's illiteracy (ß = .18; p < .001), no hand wash before eating (ß = .08; p = .004), and no hand wash after defecation (ß = .10; p < .001). This puts emphasis clearly on the awareness at household level, especially of mothers and children under age 5 in Dhaka, Bangladesh, in formulating migration-related policies.

Effectiveness of Using Mobile Phone Image Capture for Collecting Secondary Data: A Case Study on Immunization History Data Among Children in Remote Areas of Thailand.

BACKGROUND: Entering data onto paper-based forms, then digitizing them, is a traditional data-management method that might result in poor data quality, especially when the secondary data are incomplete, illegible, or missing. Transcription errors from source documents to case report forms (CRFs) are common, and subsequently the errors pass from the CRFs to the electronic database. OBJECTIVE: This study aimed to demonstrate the usefulness and to evaluate the effectiveness of mobile phone camera applications in capturing health-related data, aiming for data quality and completeness as compared to current routine practices exercised by government officials. METHODS: In this study, the concept of "data entry via phone image capture" (DEPIC) was introduced and developed to capture data directly from source documents. This case study was based on immunization history data recorded in a mother and child health (MCH) logbook. The MCH logbooks (kept by parents) were updated whenever parents brought their children to health care facilities for immunization. Traditionally, health providers are supposed to key in duplicate information of the immunization history of each child; both on the MCH logbook, which is returned to the parents, and on the individual immunization history card, which is kept at the health care unit to be subsequently entered into the electronic health care information system (HCIS). In this study, DEPIC utilized the photographic functionality of mobile phones to capture images of all immunization-history records on logbook pages and to transcribe these records directly into the database using a data-entry screen corresponding to logbook data records. DEPIC data were then compared with HCIS data-points for quality, completeness, and consistency. RESULTS: As a proof-of-concept, DEPIC captured immunization history records of 363 ethnic children living in remote areas from their MCH logbooks. Comparison of the 2 databases, DEPIC versus HCIS, revealed differences in the percentage of completeness and consistency of immunization history records. Comparing the records of each logbook in the DEPIC and HCIS databases, 17.3% (63/363) of children had complete immunization history records in the DEPIC database, but no complete records were reported in the HCIS database. Regarding the individual's actual vaccination dates, comparison of records taken from MCH logbook and those in the HCIS found that 24.2% (88/363) of the children's records were absolutely inconsistent. In addition, statistics derived from the DEPIC records showed a higher immunization coverage and much more compliance to immunization schedule by age group when compared to records derived from the HCIS database. CONCLUSIONS: DEPIC, or the concept of collecting data via image capture directly from their primary sources, has proven to be a useful data collection method in terms of completeness and consistency. In this study, DEPIC was implemented in data collection of a single survey. The DEPIC concept, however, can be easily applied in other types of survey research, for example, collecting data on changes or trends based on image evidence over time. With its image evidence and audit trail features, DEPIC has the potential for being used even in clinical studies since it could generate improved data integrity and more reliable statistics for use in both health care and research settings.

Novel mutation detection IN rpoB OF rifampicin-resistant Mycobacterium tuberculosis using pyrosequencing.

Tuberculosis (TB) remains a major global public health problem particularly severe in parts of Asia and Africa, where often it is present in HIV-AIDS patients. Although rifampicin-resistant (RIFr) TB is slow to emerge due to the low rate of mutation of its target leading to RIFE being a marker of TB that is already resistant to other anti-TB drugs, and such cases are prone to treatment failure. More than 95% of rifampicin resistance is associated with mutations in Mycobacterium tuberculosis (MTB) rpoB, with 97% of mutations occurring within the 81 bp rifampicin-resistant determining region (RRDR) of this gene. In this study, we employed pyrosequencing technique to identify mutations in RRDR and 5 codons beyond of 39 MTB strains, comprising of 14 multi-drug resistance TB (MDRTB) and 3 RIF susceptible (RIFs) MTB from the Center of Disease Control (CDC), Ratchaburi Province, and 19 mono RIFr MTB, 1 MDRTB and 2 poly-drug resistant MTB from the Chest Institute, Ministry of Public Health, Thailand. Mu- tations in 8/22 samples from the Chest Institute and 13/14 from CDC were able to be identified. Six point mutations were detected, with Ser531Leu mutation accounting for 13, the silent mutation at Gly536 for 4, deletion of Gly523 for 2, combination of His526Cys and novel Leu533Arg for 1, and a novel Leu538Arg for 1. Mutation analysis of the 81 bp fragment and 5 codons beyond in MTB rpoB using pyrosequencing provides a useful approach in predicting RIFr phenotype allowing early diagnosis and appropriate drug therapy.

Antibody germline characterization of cross-neutralizing human IgGs against 4 serotypes of dengue virus.

Dengue virus (DENV), a re-emerging virus, constitutes the largest vector-borne disease virus, with 50-100 million cases reported every year. Although DENV infection induces lifelong immunity against viruses of the same serotypes, the subsequent infection with the heterologous serotypes can cause more severe form of the disease, such as Dengue Haemorrhagic Fever (DHF) or Dengue Shock Syndrome (DSS). However, there is neither approved vaccine nor specific drugs available to treat this disease. In this study, previously developed 19 human monoclonal antibodies (HuMAbs) showing strong to moderate cross neutralizing activity were selected. Most of them (13/19) were targeted to domain II of envelop glycoprotein. To understand and clarify the recognition properties, the maturation mechanisms comprising Variable/Diversity/Joining (VDJ) recombination, Variable Heavy (VH)/Variable Light (VL) chain pairing, variability at junctional site, and somatic hypermutation (SHM) of those antibodies were studied and compared with their predecessor germline sequences. IMGT/V-QUEST database was applied to analyze the isolated VH and VL sequences. To confirm the correction of isolated VH/VL, 3 HuMAbs (1A10H7, 1B3B9, 1G7C2) was transiently expressed in HEK293T cell. All three clones of the expressed recombinant IgG (rIgG) showed the same binding and neutralizing activity as same as those from hybridomas. The data obtained in this study will elucidate the properties of those HuMAbs for further genetic modification, and its binding epitopes.

Ethical issues in research involving minority populations: the process and outcomes of protocol review by the Ethics Committee of the Faculty of Tropical Medicine, Mahidol University, Thailand.

BACKGROUND: Recruiting minorities into research studies requires special attention, particularly when studies involve "extra-vulnerable" participants with multiple vulnerabilities, e.g., pregnant women, the fetuses/neonates of ethnic minorities, children in refugee camps, or cross-border migrants. This study retrospectively analyzed submissions to the Ethics Committee of the Faculty of Tropical Medicine (FTM-EC) in Thailand. Issues related to the process and outcomes of proposal review, and the main issues for which clarification/revision were requested on studies, are discussed extensively. METHODS: The study data were extracted from proposals and amendments submitted to the FTM-EC during the period October 2009 - September 2012, and then analyzed qualitatively and quantitatively. The main issues for clarification/revision were analyzed by thematic content analysis. RESULTS: 373 proposals were submitted; 44 studies involved minority groups with 21 extra-vulnerable minorities. All clinical and 2/3 of non-clinical studies submitted for initial review underwent full-board review. For combined clinical and non-clinical study submissions, 92.1% were referred back to the investigators and approved after clarification/revision, while 2.7% were deferred due to major/critical changes, and 2.1% not approved due to substantial violations of ethical principles. The main issues needing clarification/revision differed between all studies and those involving minorities: participant information sheet (62.2% vs. 86.4%), informed consent/assent form (51.2% vs. 86.4%), and research methodology (80.7% vs. 84.1%), respectively. The main ethical issues arising during the meetings, regarding studies involving minorities, included ensuring no exploitation, coercion, or pressure on the minority to participate; methodology not affecting their legal status; considering ethnicity and cultural structure; and providing appropriate compensation. CONCLUSION: Delays in the approval or non-approval of studies involving minorities were mainly due to major or minor deviations from acceptable ethical standards and/or unclear research methodology. The FTM-EC has employed several mechanisms in its operations, including transparency in the review process, building good relationships via open communication with investigators, requesting investigators to consider closely the necessity to enroll minority groups and the risk-benefits for individuals and their communities, and the inclusion of minority-community engagement when developing the proposal. Other effective activities include annual study-site inspections, and offering refresher courses to raise awareness of minority and vulnerability issues among researchers.

Genetic variations of glutathione s-transferase influence on blood cadmium concentration.

The glutathione S-transferases (GSTs) are involved in biotransformation and detoxification of cadmium (Cd). Genetic polymorphisms in these genes may lead to interindividual variation in Cd susceptibility. The objective of this study was to assess the association of GSTs (GSTT1, GSTM1, and GSTP1 Val105Ile) polymorphisms with blood Cd concentrations in a nonoccupationally exposed population. The 370 blood samples were analyzed for Cd concentration and polymorphisms in GSTs genes. Geometric mean of blood Cd among this population was 0.46 ± 0.02 µg/L (with 95% CI; 0.43-0.49 µg/L). Blood Cd concentrations in subjects carrying GSTP1 Val/Val genotype were significantly higher than those with Ile/Ile and Ile/Val genotypes. No significant differences in blood Cd concentrations among individual with gene deletions of GSTT1 and GSTM1 were observed. GSTP1/GSTT1 and GSTP1/GSTM1 combinations showed significantly associated with increase in blood Cd levels. This study indicated that polymorphisms of GSTP1 combined with GSTT1 and/or GSTM1 deletion are likely to influence on individual susceptibility to cadmium toxicity.

Effect of crude extract of Solanum xanthocarpum against snails and mosquito larvae.

The ethanolic crude extract from Solanum xanthocarpum was investigated for its molluscicidal activity against Biomphalaria glabrata, the snail vector of Schistosoma mansoni, and Indoplanorbis exustus, the snail vector of intestinal echinostomiasis and Schistosoma spindale, together with the larvicidal activity against the larvae of Aedes aegypti, mosquito vector of dengue hemorrhagic fever and Culex quinquefasciatus, the mosquito vector of urban bancroftian filariasis. The bioassays were carried out following the methods recommended by the World Health Organization. For molluscicidal activity, the LC50 against Bi. glabrata and I. exustus were reported at 163.85 and 198.00 mg/l while the LC90 were 219.33 and 236.80 mg/l, respectively. Regarding mosquito larvicidal activity, the LC50 against the larvae of Ae. aegypti and Cx. quinquefasciatus were 788.10 and 573.20 mg/l, while the LC90 were 1288.91 and 1066.93 mg/l, respectively. These results suggest a preparation of ingredients from this plant may be used as a biological larvicide for these vectors in the field.

Gender differences in KAP related to HIV/AIDS among freshmen in Afghan universities.

This cross-sectional study aimed to describe the level of knowledge, perception/ attitude, and practices related to HIV among 1,054 freshmen students in four Afghan universities differences between genders. A probability, two stage sampling method was used. Data were collected by a self administered structured questionnaire. SPSS software was used for data analysis. Descriptive and inferential statistics were performed. Most of respondents were male (72.1%), their average age was 20.1 +/- 2 years, and most were unmarried (93.4%). The majority (90.8%) were aware of HIV but only 28.3% had a good level of knowledge. Around one-third (35.6%) had a positive level of attitude toward HIV. Approximately 30% had at least one risk practice; therefore, they were counted as high-risk behavior group members. Females were statistically more knowledgeable than males, and high-risk behaviors were significantly more prevalent among males; p = 0.01 and p = 0.001, respectively. However, general awareness, and attitude were not statistically different between genders. A considerable proportion of students (14.6%), as compared to peer-countries, were sexually active. A very high level of sharing injecting needles (4.5%) and shaving sets (20.8%) were also reported among informants.

Development of a combined air sampling and quantitative real-time PCR method for detection of Legionella spp.

The objective of this study was to develop and optimize the combined methods of air sampling and real time polymerase chain reaction (real-time PCR) for quantifying aerosol Legionella spp. Primers and TaqMan hydrolysis probe based on 5S rRNA gene specific for Legionella spp were used to amplify a specific DNA product of 84 bp. The impinger air sampler plus T-100 sampling pump was used to collect aerosol Legionella and as low as 10 fg of Legionella DNA per reaction could detected. Preliminary studies demonstrated that the developed method could detect aerosol Legionella spp 1.5-185 organisms /500 l of air within 5 hours, in contrast to culture method, that required a minimum of 7-10 days.

Preparation of in vivo cow control blood samples for cadmium analysis.

Quality control is essential for any analysis in the laboratory. The objective of this study was to prepare in vivo cow control blood samples. The experiment was performed by feeding cows with a single dose of cadmium in the form of cadmium chloride, withdrawing the blood at an appropriate time to get the highest level of cadmium and detecting the level of cadmium in the blood. It was found that feeding the cow a single dose of 0.06 mg cadmium per kg body weight resulted in the highest cadmium level of 3.622 microg/l 30-60 minutes after feeding. The samples were homogeneous because feeding the cows with single dose of cadmium let the cadmium be absorbed and distributed naturally. In addition, the samples were stable during transport. Therefore, they may be used as quality control samples to detect cadmium levels without using a lyophilized process. They could be used for proficiency testing and to evaluate whole blood analysis in the laboratory.

Determination of lead toxicity in Culex quinquefasciatus mosquitoes in the laboratory.

Laboratory investigations were carried out to study the effects of lead toxicity and lead uptake on Culex quinquefasciatus mosquitoes. Three different concentrations of lead nitrate were used in laboratory tests (0.05, 0.1, and 0.2 mg/l). An atomic absorption spectrometer (AAS) was used to the determine lead concentrations. The results showed that lead significantly reduced hatching, egg-production, and emergence rates, compared with the unexposed group (p < 0.05). The ratio of female to male offspring was 3.64:1, which was observed in the second generation, after the parents were exposed to 0.2 mg/l lead. No effects were observed on oviposition preference, larval weight, or larval deformation. The LC50 of lead against Cx. quinquefasciatus larvae within 24 hours was 0.18 mg/l. There was a significant increase in lead uptake related to increased lead exposure in mosquito larvae (p < 0.05). The bioconcentration factor (BCF) showed that the lead concentration in the larvae was 62 times greater than in the water. The lead concentration from parents to offspring reduced in the first and second generations (p < 0.05). There was no significant difference between female and male mosquitoes in lead concentration (p > 0.05).

Effect of antisense-SGTPs on the glucose uptake of the blood fluke Schistosoma mansoni: observations in adult worms and schistosomula.

AS-ODNs, complementary to Schistosoma mansoni glucose transporter proteins (SGTP1 and SGTP4), were chosen as potential therapeutic agents for schistosomiasis. AS-SGTP1 oligos lowered the glucose uptake of adult worms both in vitro and ex vivo. The most effective AS-ODN was that of 21 nucleotides complementary to the SGTP1 nucleotide sequence, including the initiation region of mRNA translation. This oligo was found to decrease glucose uptake in vitro by as much as 50% and at a concentration of 4.0 mg/ml, it killed all male worms within 24 hours. A significant decrease, up to 34%, in glucose uptake was also noted when 100 mg/kg x2 (with a 2 hours interval) of AS-ODN was administered ex vivo. Two out of six anti-SGTP4 oligos also decreased the glucose uptake of adult worms in vitro by 25-44%. Added to the culture of schistosomula, two AS-SGTP4 oligos were found to decrease glucose uptake by 20-43%.

Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water.

Multiplex PCR amplification of lacZ, uidA and plc genes was developed for the simultaneous detection of total coliform bacteria for Escherichia coli and Clostridium perfringens, in drinking water. Detection by agarose gel electrophoresis yielded a band of 876 bp for the lacZ gene of all coliform bacteria; a band of 147 bp for the uidA gene and a band of 876 bp for the lacZ gene of all strains of E. coli; a band of 280 bp for the p/c gene for all strains of C. perfringens; and a negative result for all three genes when tested with other bacteria. The detection limit was 100 pg for E. coli and C. perfringens, and 1 ng for coliform bacteria when measured with purified DNA. This assay was applied to the detection of these bacteria in spiked water samples. Spiked water samples with 0-1,000 CFU/ml of coliform bacteria and/or E. coli and/or C. perfringens were detected by this multiplex PCR after a pre-enrichment step to increase the sensitivity and to ensure that the detection was based on the presence of cultivable bacteria. The result of bacterial detection from the multiplex PCR was comparable with that of a standard plate count on selective medium (p=0.62). When using standard plate counts as a gold standard, the sensitivity for this test was 99.1% (95% CI 95.33, 99.98) and the specificity was 90.9 % (95% CI 75.67, 98.08). Multiplex PCR amplification with a pre-enrichment step was shown to be an effective, sensitive and rapid method for the simultaneous detection of these three microbiological parameters in drinking water.

Mutations in the gyrA and gyrB genes of fluoroquinolone-resistant Mycobacterium tuberculosis from TB patients in Thailand.

Among fluoroquinolone-resistant Mycobacterium tuberculosis (FQr-MTB) isolates, mutation at positions 90, 91, and 94 in gyrA gene and at positions 495, 516, and 533 in gyrB gene have been frequently reported. In this study, 35 isolates of FQr-MTB were collected from Siriraj Hospital and Chest Disease Institute. The quinolone-resistance-determining regions (QRDR) of gyrA and gyrB genes in all 35 FQr-MTB isolates and from the H37Ra MTB strain were amplified using polymerase chain reaction (PCR). DNA-sequencing and single-strand conformation polymorphism (SSCP) were further utilized for characterization of the mutations in the QRDR of gyrA and gyrB genes and mutation screening, respectively. From DNA-sequencing, 21 of 35 (60%) exhibited single-point mutations in different positions, at Ala90Val, Ser91Pro, and Asp94(Gly/Ala/His/Asn); and one novel mutation position at Gly88Cys in the gyrA gene and Asp495Asn in the gyrB gene. These positions were previously frequently reported to be responsible for FQr-MTB. The other 14 FQr-MTB isolates (40%) had no mutation. This study is the first report of mutation occurring only in the QRDR of the gyrB gene, without prior mutation in the gyrA QRDR among FQr-MTB isolates. By SSCP analysis for screening of the mutant FQr-MTB, the SSCP patterns of mutated FQr-MTB isolates were clearly differentiated from the SSCP patterns of FQs-MTB.

Simultaneous determination of trans, transmuconic acid and s-phenylmercapturic acid by high pressure liquid chromatography and its application.

The simultaneous determination of urinary trans,trans-muconic acid (t,t-MA) and S-phenylmercapturic acid (S-PMA) was performed by liquid extraction with ethyl acetate and reversed-phase high performance liquid chromatography (RP-HPLC) on a Hypersil-ODS column using the gradient mobile phase of methanol and 0.0012 N perchloric acid and diode array detection at 205 and 264 nm for S-PMA and t,t-MA, respectively. The retention times for t,t-MA and S-PMA were 3.8 and 12.3 minutes, respectively. The recoveries of t,t-MA and S-PMA were > 97%; between-day precisions were all within 8% RSD (100x SD/mean). The method was applied to analyze the urinary t,t-MA and S-PMA of 59 service station attendants exposed to average benzene concentrations in the air of 0.20+/-0.18 ppm. Significant differences in pre-shift and post-shift urinary t,t-MA between smokers and non-smokers were found.

Comparison of biological specimens for manganese determination among highly exposed welders.

This research aimed to determine if less invasive biological specimens (other than blood), such as feces and clipped toenails could be used to determine manganese concentrations among occupationally exposed human subjects. In addition to blood samples, which have routinely been used in determining manganese concentration, specimens were collected from welders working at the Electricity Generating Authority of Thailand, Mae Moh Thermal Power Plant, Lampang Province. Manganese concentrations in these three biological samples were determined by atomic absorption spectrophotometer. Correlations of manganese concentrations among these three biological samples were measured, and found to be rather poor (Pearson's r <+/-0.2, p > 0.1 for any pair-wise comparisons). Blood remains the recommended material for biomonitoring manganese concentrations in occupationally exposed subjects.