Investigation of an outbreak of anemia cases at an Army trinitrotoluene munitions production plant from 2004 to 2005 and subsequent surveillance 2005-2013.

OBJECTIVE: Investigate an outbreak of anemia that occurred at an Army trinitrotoluene (TNT) munitions production facility in 2004-2005 and describe subsequent medical surveillance efforts between 2005 and 2013. METHODS: A retrospective, cross-sectional investigation of anemia cases and exposure levels was conducted to target corrective engineering and administrative changes needed to reduce TNT levels and anemia cases. RESULTS: Mean TNT exposure levels decreased from 1.2 to 0.08 mg/m(3) between 2004 and 2012, but the trend was not statistically significant. Anemia cases rose from 17 to 38 between 2004 and 2011 and cases declined from 4 to 0 between 2012 and 2013. The overall trend in cases was not statistically significant. CONCLUSIONS: TNT levels rose at a munitions plant until ventilation changes reduced exposures. Despite this, anemia cases continued to rise until the commander held workers accountable for failing to wear PPE and motivated them to wear respirators to avoid a loss in pay. The case definition of anemia was revised to be consistent with the case definition used by private physicians who practiced in the community. Better coveralls are needed, which are easy to wear, reduce static discharge, and prevent TNT skin absorption.

2,4,6-Trinitrotoluene (TNT) air concentrations, hemoglobin changes, and anemia cases in respirator protected TNT munitions demilitarization workers.

PURPOSE: 2,4,6-Trinitrotoluene (TNT) is an explosive used in munitions production that is known to cause both aplastic and hemolytic anemia in exposed workers. Anemia in a TNT worker is considered a sentinel health event (occupational) (SHE(O)) in the United States (US). Deaths have been reported secondary to aplastic anemia. Studies have shown that TNT systemic absorption is significant by both the respiratory and dermal routes. No studies encountered looked at hemoglobin change or anemia cases in respiratory protected workers. It is hypothesized that respiratory protection is insufficient to protect TNT workers from the risk of anemia development and hemoglobin concentration drop. METHODS: A records review of eight groups of respiratory protected TNT workers' pre-exposure hemoglobin levels were compared with their during-exposure hemoglobin levels for statistically significant (alpha level 0.05) hemoglobin level changes, and anemia cases were recorded. A curve estimation analysis was performed between mean TNT air concentrations and mean hemoglobin change values. RESULTS: Statistically significant hemoglobin level drops and anemia cases were apparent at TNT air concentrations about the REL and PEL in respiratory protected workers. There were no anemia cases or statistically significant hemoglobin level drops at concentrations about the TLV, however. A statistically significant inverse non-linear regression model was found to be the best fit for regressing hemoglobin change on TNT air concentration. CONCLUSIONS: Respiratory protection may be inadequate to prevent workers who are at risk for TNT skin absorption from developing anemia. This study contributes evidence that the TLV should be considered for adoption as the new PEL.

Biomonitoring of workers cleaning up ammunition waste sites.

2,4,6-Trinitrotoluene (TNT) is an important occupational and environmental pollutant. In TNT-exposed humans, notable toxic manifestations have included aplastic anaemia, toxic hepatitis, cataracts, hepatomegaly and liver cancer. Therefore, it is important to develop protection measures and to monitor workers involved in the clean-up of ammunition sites. Haemoglobin (Hb) adducts of TNT, 4-amino-2,6-dinitrotoluene (4ADNT) and 2-amino-4,6-dinitrotoluene (2ADNT), and the urine metabolites of TNT, 4ADNT and 2ADNT were found in 22-50% of the exposed workers, but not in the control group. The exposed workers were wearing protective equipment. The levels of erythrocytes, haemoglobin, creatinine, serum glutamic pyruvic transaminase and lymphocyte levels were significantly lower in the exposed workers than in the non-exposed workers. The levels of blood urea and reticulocytes were significantly higher in the exposed workers than in the non-exposed workers. Headache (26%), mucous membrane irritation (16%), sick leave (18%), lassitude (8%), anxiety (6%), shortness of breath (3%), nausea (5%) and allergic reactions (8%) were reported by the exposed workers. In a further analysis the U-4ADNT levels and the Hb-adduct levels were compared to the blood parameter and the health effects. The blood parameters were not significantly different between the U-4ADNT positive and U-4ADNT-negative group. Headache, mucous membrane irritation, sick leave, lassitude, anxiety, shortness of breath and allergic reactions were statistically not different between the two groups. Also in the workers with Hb-4ADNT adducts no significant negative changes were seen in regards to the changes of the blood parameters or the health effects. According to the results of the present study, it appears that the blood parameter changes and the health effects are more influenced by other factors than by the internal exposure to TNT.

Analytical monitoring of trinitrotoluene metabolites in urine by GC-MS. Part I. Semiquantitative determination of 4-amino-2,6-dinitrotoluene in human urine.

A routine procedure for the monitoring of occupational exposure to 2,4,6-trinitrotoluene (TNT) based on the semiquantitative determination of 4-amino-2,6-dinitrotoluene (4A2,6-DNT) in urine samples by gas chromatography (GC-MS) was developed. For calibration purposes, a standard sample of 4-amino-4,6-dinitrotoluene was prepared. Urine samples were collected from munition workers in a military plant in Hradec Kralove district. The samples were extracted into ethyl acetate, and analyzed by GC-MS without previous derivatization. The GC was equipped with a DB1701 column (30 m, 0.25 mm, 0.25 microm) for optimum separation. 4A2,6-DNT was detected at concentrations above the limit of quantitation (0.2 mg/L). This simple method is sufficient for the screening of high degree of TNT exposure and constitutes the first step of a broad project focused on this occupational medicine problem.

Trinitrotoluene (TNT)-induced cataract in Danish arms factory workers.

PURPOSE: To compare the prevalence of cataract in workers exposed to trinitrotoluene (TNT) to the prevalence in a group of unexposed workers, matched on age and sex, using Tiukina's description and grading of TNT-induced cataract. METHODS: A total of 23 TNT-exposed and 44 unexposed workers underwent an eye examination performed by an ophthalmologist who did not know the exposure status of the subjects. All lens opacities matching Tiukina's description were classified as TNT cataract and graded on Tiukina's scale of stages 1-4. RESULTS: Four cases of TNT-induced cataract were identified among the 23 TNT-exposed workers and none in the unexposed group (p < 0.01). CONCLUSION: Exposure to TNT may cause a unique type of cataract, which a general ophthalmologist, using Tiukina's description and grading scale, will be able to distinguish from other cataracts.

[Complex evaluation of health in patients with occupational eye pathology]. / Kompleksnaia otsenka zdorov'ia bol'nykh s professional'noi patologiei organa zreniia.

The article deals with data on epidemiology of occupational eye diseases in underground shotfirers. The author suggests method for complex evaluation of their health state.

Military explosives and health: organic energetic compound syndrome?

Military explosives, which are Organic Energetic Compounds (OECs), might be initiating factors in clusters of warfare related illnesses. Substances such as HMX, RDX and TNT are neurotoxic and are possible human carcinogens. Not all the explosive material is consumed in an explosion so that the use of explosives generates harmful dust. In many clusters of illnesses a link can be defined with explosives. In researching for the cause of the illnesses several possibilities are mentioned, but explosives are rarely mentioned. This article considers the possible role of OECs, with the intention of encouraging studies to make the role of explosives clear: no cause, a link with insufficient evidence or the cause of many illnesses.

Exposure to nitroaromatic explosives and health effects during disposal of military waste.

AIMS: To investigate the exposure to dinitrotoluene (DNT) and trinitrotoluene (TNT) and the resulting effects in workers which occur during the disposal of military waste. METHODS: Eighty two employees from a mechanical plant in Germany were studied, of whom 51 were regularly exposed to ammunition containing TNT and DNT, 19 occasionally, and 12 not at all. RESULTS: Air analyses yielded maximum concentrations of 20 micro g/m(3) for 2,4-DNT and 3250 micro g/m(3) for 2,4,6-TNT, respectively. The maximum concentrations in the urine of workers regularly exposed amounted to 5.0 micro g/l of 2,4,6-TNT, 1464.0 micro g/l of 2-amino-4,6-dinitrotoluene, 6693.0 of micro g/l 4-amino-2,6-dinitrotoluene, 2.1 micro g/l of 2,4-DNT, 95.0 micro g/l of 2,4-dinitrobenzoic acid, and 3.6 micro g/l of 2,6-DNT. There was a highly significant linear correlation between the urinary concentrations of the two main metabolites of TNT, 2-amino-4,6-dinitrotoluene and 4-amino-2,6-dinitrotoluene. In 63 persons TNT or DNT or metabolite concentrations above the analytical detection limit were found in urine. These persons reported more frequently symptoms like bitter taste, burning eyes, and discoloration of the skin and hair than persons (n = 19) without detectable TNT and/or DNT exposure. CONCLUSION: During the disposal of military waste containing relevant TNT and DNT, exposure can occur of occupational-medical relevance. Biological monitoring is suitable for the early detection of possible adverse effects at workplaces exposed to TNT. Protective measures should be improved, together with adequate occupational-medical surveillance of persons exposed to nitroaromatic explosives. Further studies are necessary to exclude possible long term effects.

TNT, RDX, and HMX decrease earthworm (Eisenia andrei) life-cycle responses in a spiked natural forest soil.

Sublethal and chronic toxicities of 2,4,6-trinitrotoluene (TNT), 1,3,5-trinitro-1,3,5-triazacyclohexane (RDX), and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) on earthworm Eisenia andrei in a sandy forest soil were assessed. Various reproduction parameters of fecundity (total and hatched number of cocoons, number of juveniles, and their biomass) were significantly decreased by TNT (> or = 58.8 +/- 5.1 mg/kg dry soil), RDX (> or = 46.7 +/- 2.6 mg/kg), and HMX (> or = 15.6 +/- 4.6 mg/kg). These effects occurred at much lower concentrations than those reported earlier using artificial soil preparations. Growth of adults was significantly decreased in the TNT-spiked natural soils at 136.2 +/- 25.6 mg/kg dry soil, the highest concentration having no significant mortality. In contrast, survival and growth were not significantly reduced at relatively high measured concentrations of RDX (167.3 mg/kg) and HMX (711.0 mg/kg). Although TNT, RDX, and HMX share a common life-cycle response ( i.e., decreased juvenile counts), a number of differences related to other reproduction parameters (e.g., productivity of cocoons) was observed. These results indicate that the tested explosives do not support a common mechanism of toxicity, at least in the earthworm, probably due to differences in their physical-chemical properties as well as metabolites formed during exposure.

Biological remediation of explosives and related nitroaromatic compounds.

Nitroaromatics form an important group of recalcitrant xenobiotics. Only few aromatic compounds, bearing one nitro group as a substituent of the aromatic ring, are produced as secondary metabolites by microorganisms. The majority of nitroaromatic compounds in the biosphere are industrial chemicals such as explosives, dyes, polyurethane foams, herbicides, insecticides and solvents. These compounds are generally recalcitrant to biological treatment and remain in the biosphere, where they constitute a source of pollution due to both toxic and mutagenic effects on humans, fish, algae and microorganisms. However, relatively few microorganisms have been described as being able to use nitroaromatic compounds as nitrogen and/or carbon and energy source. The best-known nitroaromatic compound is the explosive TNT (2,4,6-trinitrotoluene). This article reviews the bioremediation strategies for TNT-contaminated soil and water. It comes to the following conclusion: The optimal remediation strategy for nitroaromatic compounds depends on many site-specific factors. Composting and the use of reactor systems lend themselves to treating soils contaminated with high levels of explosives (e.g. at former ammunition production facilities, where areas with a high contamination level are common). Compared to composting systems, bioreactors have the major advantage of a short treatment time, but the disadvantage of being more labour intensive and more expensive. Studies indicate that biological treatment systems, which are based on the activity of the fungus Phanerochaete chrysosporium or on Pseudomonas sp. ST53, might be used as effective methods for the remediation of highly contaminated soil and water. Phytoremediation, although not widely used now, has the potential to become an important strategy for the remediation of soil and water contaminated with explosives. It is best suited where contaminant levels are low (e.g. at military sites where pollution is rather diffuse) and where larger contaminated surfaces or volumes have to be treated. In addition, phytoremediation can be used as a polishing method after other remediation treatments, such as composting or bioslurry, have taken place. This in-situ treatment method has the advantage of lower treatment costs, but has the disadvantage of a considerable longer treatment time. In order to improve the cost-efficiency, phytoremediation of nitroaromatics (and other organic xenobiotics) could be combined with bio-energy production. This requires, however, detailed knowledge on the fate of the contaminants in the plants as well as the development of efficient treatment methods for the contaminated biomass that minimise the spreading of the contaminants into the environment during post harvest treatment.

Exposure to armament wastes and leukemia: a case-control study within a cluster of AML and CML in Germany.

An unusually high incidence of acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) concentrated in a specific locality of a region in Germany motivated a descriptive incidence study in that region which showed a near 10-fold increased risk of CML among males but not among females (Kolb G, Becker N, Scheller S, Zugmaier G, Pralle H, Wahrendorf J, Havemann K. Increased risk of acute myelogenous leukemia (AML) and chronic myelogenous leukemia (CML) in a County of Hesse, Germany, Soc Prev Med 1993;38:190-195). Since a serious environmental contamination of areas in this locality with armament wastes containing toluene-derivatives has been known for a long time, the hypothesis arose that TNT production and the related severe contamination of soil and water might be responsible for the observed higher risk. We carried out a case-control study within the cluster to test this hypothesis. Overall, the results do not confirm the hypothesis. There is an indication of a relationship of an increased odds ratio with the exposure for a small group of persons who lived at a particular site in one of the two communities involved during the peak phase of TNT production during the 1940s. However, this finding is spurious and cannot explain the large majority of cases which occurred in that area in the 1980s. At the moment, no other explanation can be given for the increased risk of leukemias in that area.

Transcriptional activation of stress genes and cytotoxicity in human liver carcinoma cells (HepG2) exposed to 2,4,6-trinitrotoluene, 2,4-dinitrotoluene, and 2,6-dinitrotoluene.

The CAT-Tox (L) assay has recently been developed and validated for detecting and quantifying the specific molecular mechanisms that underlie toxicity of various xenobotic chemicals. We performed this assay to measure the transcriptional responses associated with 2,4,6-trinitrotoluene (TNT) and 2 of its byproducts [2,4 and 2,6-dinitotoluenes (DNTs)] to 13 different recombinant cell lines generated from human liver carcinoma cells (HepG2) by creating stable transfectants of mammalian promoter chloramphenicol acetyltransferase (CAT) gene fusions. Cytoxicity test with the parental HepG2 cells, using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]-based assay for cell viability, yielded LC50 values of 105 +/- 6 mg/mL for TNT in 1% dimethyl sulfoxide (DMSO), and > 300 mg/mL for DNTs, upon 48 h of exposure. TNT appeared to be more toxic than 2,4-DNT, which also showed a higher toxicity compared to 2,6-DNT. Of the 13 recombinant constructs evaluated, 8 (CYP 1A1, GST Ya, XRE, HMTIIA, c-fos, HSP70, GADD153, and GADD45), 5 (c-fos, HSP70, GADD153, GADD45, and GRP78), and none showed inductions to significant levels (p < 0.05), for TNT, 2,4-DNT, and 2,6-DNT, respectively. For most constructs, the induction of stress genes was concentration-dependent. These results show the potential for TNT and 2,4-DNT to cause protein damage and/or perturbations of protein biosynthesis (HSP70 and GRP78), alterations in DNA sequence or its helical structure (c-fos, GADD153, GADD45), and the potential involvement of TNT in the biotransformation process (CYP 1A1, GST Ya, XRE), and in the toxicokinetics of metal ions (HMTIIA). Within the range of concentrations tested (0-300 mg TNT or DNT/mL in 1% DMSO), no significant inductions (p > 0.05) of NFKBRE, p53RE, CRE, and RARE were found.

Lens opacifications detected by slitlamp biomicroscopy are associated with exposure to organic nitrate explosives.

CONTEXT: Unusual cataracts (flecks) have been reported to occur at very low levels of trinitrotoluene exposure, but prevalence estimates vary widely. Cataracts have not been reported among workers in the United States exposed to organic nitrate explosives. OBJECTIVES: To determine the prevalence of unusual cataracts in a population of workers in the United States exposed to organic nitrate explosives, to determine whether associations exist with reported cataract risk factors, and to determine if other eye effects (eg, retinal hemorrhage) are associated with exposure. DESIGN: Cohort prevalence study. SETTING: A university-based ophthalmologic clinic. SUBJECTS: Sixty-one workers from an explosives plant comprised the exposed group. The comparison group consisted of 56 workers using chemicals other than organic nitrate explosives. OUTCOME MEASURES: The primary outcome measure was opacifications (flecks) of the crystalline lens, graded clinically on a scale of 0 to 4 +. Additional measures included visual acuity, applanation tonometry, and clinical evaluation using standard examination techniques. RESULTS: Sixty-three percent of the workers had anterior cortical lens opacifications in a pattern of peripheral flecks. Exposed subjects were 18 times more likely to exhibit changes than those not exposed, a statistically significant association (95% confidence interval [CI], 5.0-65.0; P<.001). A statistically significant association with the duration of exposure was also found. CONCLUSIONS: Asymptomatic, low-grade cataracts (flecks) were identified in 63% of the workers exposed to pentolite. No other eye effects were found to be associated with exposure. Cataracts were not associated with other known risk factors, but were associated with the duration of exposure. Biomicroscopy is widely available and useful for detecting changes in the asymptomatic stages.

An in situ respirometric technique to measure pollution-induced microbial community tolerance in soils contaminated with 2,4, 6-trinitrotoluene.

Long-term exposure to 2,4,6-trinitrotoluene (TNT) can induce changes in the structure and activities of soil microbial communities. Such changes may be associated with an elevated microbial tolerance. An in situ respirometry technique based on the analysis of the substrate-induced respiration response to freshly added TNT was used to examine soil microbial tolerance to TNT at the community level. The specific growth rate derived by fitting an exponential equation to respiration data was taken as the measurement endpoint. Microbial tolerance was evaluated using a tolerance index defined as the ratio of the specific growth rate at a spiking dose of 2000 microg TNT/g soil to that of the control with no spiked TNT. Three soils with long-term exposure histories (TNT level in soil: 1.5, 32, and 620 microg TNT/g, respectively) exhibited significantly higher microbial community tolerance to TNT than two uncontaminated control soils. A soil containing 29,000 microg TNT/g exhibited the highest tolerance. Findings from this study support the hypothesis that pollution-induced community tolerance can be used as a means of identifying those compounds that have exerted selective pressure on the community.

Hemolytic crisis in G6PD-deficient individuals in the occupational setting.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency affects an estimated 400 million people. Most of the individuals affected live in Africa, the Middle East and Southeast Asia, or are African-Americans. The ethnic distribution of G6PD-deficiency is extremely variable in Israel--where the Jewish population has been repatriated from more than 60 different countries--its prevalence being from 0% to 58%. Five cases of hemolytic episodes arising from the loading of trinitrotoluene (TNT) were detected. Hemoglobin levels dropped between 4 and 10.2 g/dl. Reticulocytosis reached 4%-26.2% of red blood cells, while bilirubin levels rose to a peak of 5.1 mg/dl. Methemoglobin was 1.5%-8.6% and sulfmethemoglobin 0.9%-4.93% of total hemoglobin. Urinary urobilinogen was increased in all. All five workers showed absolute G6PD deficiency. In one worker a urinary metabolite of TNT 2-6 dinitromono amino toluene, was detected.

[The impact of trinitrotoluene on eyes in miners]. / Vozdeistvie trinitrotoluola na glaza shakhterov.

Trinitrotoluene (TNT) effect on the eyes of 250 miners was studied. The length of service of the investigated group varied from one to twenty years. The specific trinitrotoluene cataract of various stages was observed in 54.7% of the examinees. The authors assume that the lens changes depend on the length of service and on the TNT concentrations in the body after predominant skin penetration.

Some altered concentrations of elements in semen of workers exposed to trinitrotoluene.

OBJECTIVES: A cross sectional study was performed to find the concentrations of elements contained in the semen of workers exposed to trinitrotoluene (TNT). SUBJECTS AND METHODS: Semen of exposed workers in two TNT plants located in He-Nan Province in 1992 were examined. RESULTS: The average TNT concentrations in the workplace, except the packing site, were found to have exceeded the maximal allowable concentration (MAC, 1 mg/m3); skin contaminations of male workers exposed to TNT were higher after a shift than in controls, and correlated with the total blood concentrations of TNT, 4-amino-2, 6-dinitrotoluene (4A), and 2-amino-4, 6-dinitrotoluene (2A). Cu, Zn, Na, Mg, and Se concentrations were significantly decreased, but K, Ca, Co, Mn and Li contents were not significantly changed in the semen of workers exposed to TNT. Compared with the control group, the percentage of liquifying time of semen, the sperm malformation incidence, and viability in the men exposed to TNT were all significantly changed. CONCLUSIONS: Men exposed to TNT have decreased concentrations of some elements is semen and altered semen physiology.

[Occupational disease among miners of the Kursk Magnetic Anomaly (from the experience of the work of the Institute of Industrial Hygiene and Occupational Diseases in Heavy Metallurgy)]. / Professional'naia zabolevaemost' sredi rabochikh gornorudnykh predpriiatii Kurskoi magnitnoi anomalii (Iz opyta raboty instituta--kuratora po problemam gigieny truda i professional'nykh zabolevanii v chernoi metallurgii.

The materials are first to present analysis of occupational morbidity among miners of variable age, length of service, various occupations, who work at Kursk Magnetic Anomaly. The recommendations on lower occupational morbidity are presented.