The acute and long-term effects of Middle East sand particles on the rat airway following a single intratracheal instillation.

Military personnel deployed in the Middle East have emphasized concerns regarding high levels of dust generated from blowing desert sand and the movement of troops and equipment. Airborne particulate matter levels (PM(10); PM < 10 µm) in the region may exceed 1500 µg/m(3), significantly higher than the military exposure guideline (MEG) of 50 µg/m(3). Increases in PM(10) have been linked to a rise in incidences of asthma, obstructive pulmonary disease, lung cancer, and cardiovascular diseases. Male Sprague-Dawley rats received a single intratracheal (IT) instillation of 1, 5, or 10 mg of Middle East PM(10) collected at a military occupied site in Kuwait, silica (positive control), or titanium dioxide (TiO(2); negative control) suspended in 400 µl sterile saline, or saline alone (vehicle control). Twenty-four hours, 3 d, 7 d and 6 mo postexposure (n = 15/group), organs including lung were evaluated for histopathological changes and for particle contaminants. Bronchoalveolar fluid (BALF) was also analyzed for cellular and biochemical parameters, including cytokines and chemokines. Instillation of silica resulted in early, pronounced, sustained inflammation indicated by significant increases in levels of total protein and neutrophils, and activities of lactate dehydrogenase activity and ß-glucuronidase activity. Lower magnitude and transient changes using the same markers were observed in animals exposed to TiO(2) and Middle East PM(10). The results suggest that for acute exposures, this Middle East PM(10) is a nuisance-type dust with relatively low toxicity. However, since average deployment of military personnel to the Middle East is 180 d with potential for multiple follow-on tours, chronic exposure studies are needed to fully understand the pulmonary effects associated with Middle East PM exposure.

Two-generation reproductive toxicity study of implanted depleted uranium (DU) in CD rats.

Depleted uranium (DU) munitions and armor plating have been used in several conflicts over the last 17 yr, including the Persian Gulf War and the Iraq War. Because of its effectiveness and availability, DU will continue to be used in military applications into the foreseeable future. There is much controversy over the use of DU in weapons and equipment because of its potential radiological and toxic hazards, and there is concern over the chronic adverse health effects of embedded DU shrapnel in war veterans and bystanders. This study evaluated the effects of long-term implantation of DU on the reproductive success of F0 generation adults and development and survival of subsequent F1 and F2 generations in a two-generation reproductive toxicity study. F0 generation Sprague-Dawley rats, 8 wk of age, were surgically implanted with 0, 4, 8, 12, or 20 DU pellets (1 x 2 mm). Inert implant control animals were implanted with 12 or 20 tantallum (Ta) pellets. The F0 generation was then mated at 120 d post DU implantation. In the F0 generation, when measured on postimplantation d 27 and 117, uranium was present in the urine of DU-implanted animals in a dose-dependent manner. F0 reproductive success was similar across treatment groups and the maternal retrieval test revealed no changes in maternal behavior. DU implantation exerted no effect on the survival, health, or well-being of the F0 generation. Necropsy results of F0 animals were negative with the exception of a marked inflammatory response surrounding the implanted DU pellets. For the F1 generation, measures of F1 development through postnatal day (PND) 20 were unremarkable and no gross abnormalities were observed in F1 offspring. No uranium was detected in whole-body homogenates of PND 4 or PND 20 pups. Necropsy findings of F1 PND 20 pups were negative and no instances of ribcage malformation were observed in F1 PND 20 pups. Body weight and body weight gain of F1 rats through PND 120 were similar across treatment groups. Eight of 414 F1 animals observed from PND 20 to 120 died of unknown causes; 7 were from litters of DU-implanted F0 mating pairs. F1 mating success at 10 wk of age was an overall 70% compared with 91% for F0 mating pairs. Mating success was similar between F1 animals derived from DU-implanted F0 adults and those derived from F0 implant control adults suggesting that the comparatively low mating success was not due to F1 DU exposure. The gestational index of F1 animals derived from mid-dose F0 mating pairs was found to be lower compared with F1 controls. The average gestation duration of F1 animals derived from high-dose F0 mating pairs was found to be significantly longer than F1 controls. F1 sperm motility analyses did not differ among experimental groups and no gross abnormalities were identified at necropsy among surviving F1 animals at PND 120. Histopathology of kidneys, spleen, thymus, bone marrow, ovaries, and testes of F1 high-dose animals did not differ from F1 controls. F1 high-dose females had significantly higher mean relative liver and heart weights compared with F1 controls; the biological relevance of this finding could not be determined. For the F2 generation, measures of F2 development through PND 20 were unremarkable and no gross abnormalities were observed in F2 offspring. Necropsy findings of F2 PND 20 pups were negative and no instances of ribcage malformation were observed in F2 PND 20 pups. Body weight and body weight gain of F2 rats through PND 90 were similar across treatment groups. Mean relative heart weights of males derived from high-dose F0 parents were significantly lower compared with F2 controls. Sperm motility and concentration analysis of F2 males at PND 90 were similar across F2 groups. Overall, the consistent absence of positive findings in this study seems to suggest that DU is not a significant reproductive or developmental hazard, particularly when one considers that mid- and high-dose rats were implanted with the equivalent of 0.3 and 0.5 lb of DU in a 70-kg human, respectively. However, the findings that seven of eight F1 adults that died postweaning were from DU-implanted F0 mating pairs, and that mean relative heart weights were elevated in high-dose F1 and F2 pups, suggest conservatism is warranted in characterizing the reproductive and teratogenic hazards of embedded DU until further studies are completed.

Evaluation of the effect of implanted depleted uranium (DU) on adult rat behavior and toxicological endpoints.

In 2002, the Naval Health Research Center Toxicology Detachment began a study to determine the effects of surgically implanted depleted uranium (DU) pellets on adult rat (e.g., P1 generation) health and reproduction. In this report, the effect of implanted DU on adult rat behavior and health is described. Adult Sprague-Dawley (SD) rats, 8 wk of age, were surgically implanted with 0, 4, 8, 12, or 20 DU pellets (1 x 2 mm); 20 DU pellets of size 1 x 2 mm approximates to 0.22 kg (0.5 lb) of DU in a 70-kg (154 lb) person. Control animals were implanted with 12 or 20 tantallum (Ta) pellets. The animals were then housed for up to 150 d postimplantation or 20% of an assumed 2-yr life span for rats. The concentration of uranium in urine directly correlated with the number of implanted DU pellets, indicating that DU was migrating into the body from the implanted pellets. Three male and 4 female animals died during the 150-d period of causes apparently not related to DU implantation. Behavioral testing found no definitive evidence of neurobehavioral perturbations associated with DU implantation. Uranium translocated to tissues known to sequester uranium (bone, teeth, and kidneys), but uranium concentrations varied considerably within each dose group and did not follow a dose-response pattern as anticipated. Serum chemistry values were within normal ranges for the SD rat. However, alanine aminotransferase measurements were significantly lower for rats implanted with 20 DU pellets as compared to sham surgery controls but not when compared to animals implanted with Ta pellets only. Phosphate measurements were significantly lower for female rats implanted with 20 DU pellets as compared to both sham surgery controls and animals implanted with Ta pellets only. Monocyte ratios were higher in adult rats implanted with 20 DU pellets as compared to sham surgery controls but not when compared to animals implanted with 20 Ta pellets. Mean platelet volume was found to be significantly lower for rats implanted with 20 DU pellets as compared to sham surgery controls but not when compared to animals implanted with 20 Ta pellets. Gross necropsy found no obvious tissue abnormalities in implanted rats, and the weights of major tissues did not differ between Ta- and DU-implanted animals. Histopathologic analysis of major tissues from animals implanted with 0 pellets, 20 Ta pellets, or 20 DU pellets found no differences between treatment groups. The findings of this study indicate that implantation of up to 20 DU pellets in adult rats did not have a significant negative impact on their general health and neurobehavioral capacities when assessed after 150 d of pellet implantation. However, the growing body of data on the potential health effects associated with DU exposure warrants further studies involving higher embedded DU body burdens in conjunction with longer surveillance periods postimplantation.

Hydrocarbon-based weapons maintenance compounds produce evidence of contact hypersensitivity in BALB/c mice.

Unprotected dermal contact with weapons maintenance materials is highly probable during cleaning and maintenance of firearms. Several weapons maintenance materials of interest to the Department of Defense were evaluated for their irritating and sensitizing potential in a modified local lymph node assay (LLNA). Female BALB/c mice (n = 5) were topically exposed to Break-Free CLP, Royco 634, TW-25B, MC-25, or MC-2500. All compounds tested produced a positive response for irritancy and lymphocyte proliferation. Break-Free CLP and Royco 634 produced the greatest dermal irritation and highest LLNA stimulation index. Phenotyping of draining lymph node cells from animals treated with Break-Free CLP suggest that this material induces T-cell-mediated contact sensitization (Type IV hypersensitivity) in mice. These findings support the recommendation that persons handling or using weapons maintenance materials should protect their skin from repeated contact by wearing appropriate personal protective equipment.

Acute and subacute dermal toxicity of Break-Free CLP: a weapons cleaning and maintenance compound.

Break-Free CLP((R)) is a commercial cleaning, lubricating and preserving compound used in both the military and civilian sectors for maintenance of small- and large-caliber weapons. Like many commercial mixtures, there is very little information available on the toxicity of Break-Free CLP. Studies were conducted to characterize the biological effects of single or repeat dermal application of Break-Free CLP to the clipped backs of CD-1 mice. Break-Free CLP was applied neat, 50 microl three times of week for up to 2 weeks. Foci of epithelial ulceration were observed in skin sections from 22% of Break-Free CLP-treated animals in conjunction with markedly thickened epithelium suggesting that robust epithelial regeneration was occurring in these animals. Skin histopathology of Break-Free CLP-treated animals closely matched the histopathology from mice treated repeatedly with 2% croton oil in acetone (dermal irritation positive control). Serum alkaline phosphatase activity was significantly (P < 0.05) lower for mice treated with Break-Free CLP, 2% croton oil or 7,12-dimethylbenz[a]anthracene (DMBA) compared with negative and vehicle control mice. Skin nitric oxide (NO) levels were not significantly elevated for mice treated with Break-Free CLP but were significantly elevated for mice treated with dermal irritation positive control compound DMBA. The cumulative skin changes in Break-Free CLP-treated animals support conducting a subchronic dermal application study. The observed decreases in serum alkaline phosphatase activity suggest that future studies should include the liver and bone as possible target organs. Additionally, dermal penetration studies could provide key health risk assessment information for characterizing the potential health risks associated with chronic dermal exposure to Break-Free CLP.

Study of the reproductive effects in rats surgically implanted with depleted uranium for up to 90 days.

In 2001, the Naval Health Research Center Toxicology Detachment was funded by the U.S. Army Medical Research Acquisition Activity (USAMRAA) to conduct a study of the effects of surgically implanted depleted uranium (DU) pellets on adult rat reproductive success and development across two successive generations. This article presents some of the findings for the group of offspring from adult rats mated at 30 d post surgical implantation of DU pellets. Adult male and female Sprague-Dawley rats (P1 generation) were surgically implanted with 0, 4, 8, or 12 DU pellets (1 x 2 mm). The P1 generation was then cross-mated at 30 d post surgical implantation. Urine collected from P1 animals at 27 d post surgical implantation showed that DU was excreted in the urine of DU-implanted animals in a dose-dependent manner. DU surgical implantation did not have a negative impact on P1 reproductive success, survival, or body weight gain through post surgical implantation d 90. There were no statistically significant differences in F1 birth weight, survival, and litter size at postnatal day (PND) 0, 5, and 20. No gross physical abnormalities identified in the offspring were attributable to neonatal DU exposure. A series of neurodevelopment and immune function assessments were also conducted on F1 offspring. No group differences were observed that were related to parental DU exposure. Studies are ongoing on the impact of leaving DU embedded in soft tissue for 120 d on rat reproduction and subsequent offspring survival and development.

Estimation of the health risks associated with polychlorinated biphenyl (PCB) concentrations found onboard older U.S. Navy vessels.

PCBs have been identified on surfaces and in component materials and equipment from inactive U.S. Navy nuclear submarines commissioned prior to 1970. Health risks associated with PCBs present onboard submarines were estimated for hypothetical crew members and shipyard workers. Median non-cancer hazard quotients for shipyard workers and submarine crew ranged between 0.4-54.6, with the highest quotients estimated for unprotected shipyard workers. Median cancer risk estimates ranged from 7.3 x 10(-6) to 1.1 x 10(-3) with the highest estimated risk calculated for unprotected shipyard workers. Our findings suggest that PCB surface concentrations found onboard inactive nuclear submarines commissioned prior to 1970 may be high enough to constitute a possible risk to the health of persons involved in dismantling of Navy submarines if PCB exposure is not minimized. Potential sources of uncertainty in our risk assessment include the correlation between PCB contamination levels on inactive versus active nuclear submarine vessels, the relationship between wipe sample concentrations and human exposure, dermal contact frequency with PCB-contaminated surfaces, carcinogenicity of PCBs in humans, and uncertainties inherent with the PCB cancer slope factor and oral RfD. Our findings support Navy policy that shipyard workers should wear personal protective equipment when PCB contamination is suspected or has been identified and that IH surveys should continue to identify sources of PCB contamination onboard vessels and reduce PCB contamination to concentrations that are reasonably achievable.

A review of the effects of uranium and depleted uranium exposure on reproduction and fetal development.

Depleted uranium (DU) is used in armor-penetrating munitions, military vehicle armor, and aircraft, ship and missile counterweighting/ballasting, as well as in a number of other military and commercial applications. Recent combat applications of DU alloy [i.e., Persian Gulf War (PGW) and Kosovo peacekeeping objective] resulted in human acute exposure to DU dust, vapor or aerosol, as well as chronic exposure from tissue embedding of DU shrapnel fragments. DU alloy is 99.8% 238Uranium, and emits approximately 60% of the alpha, beta, and gamma radiation found in natural uranium (4.05 x 10(-7) Ci/g DU alloy). DU is a heavy metal that is 160% more dense than lead and can remain within the body for many years and slowly solubilize. High levels of urinary uranium have been measured in PGW veterans 10 years after exposure to DU fragments and vapors. In rats, there is strong evidence of DU accumulation in tissues including testes, bone, kidneys, and brain. In vitro tests indicate that DU alloy may be both genotoxic and mutagenic, whereas a recent in vivo study suggests that tissue-embedded DU alloy may be carcinogenic in rats. There is limited available data for reproductive and teratological deficits from exposure to uranium per se, typically from oral, respiratory, or dermal exposure routes. Alternatively, there is no data available on the reproductive effects of DU embedded. This paper reviews published studies of reproductive toxicity in humans and animals from uranium or DU exposure, and discusses ongoing animal research to evaluate reproductive effects in male and female rats embedded with DU fragments, and possible consequences in F1 and F2 generations.

The effects of near ultraviolet radiation on the toxic effects of polycyclic aromatic hydrocarbons in animals and plants: a review.

Polycyclic aromatic hydrocarbons (PAHs) are common contaminants of terrestrial and aquatic ecosystems. Traditional, toxicological studies for defining the potential hazard of PAHs in wildlife have been conducted in the absence of UV radiation. However, recent toxicology studies particularly in the discipline of aquatic toxicology have presented evidence that PAHs may become toxic or substantially more toxic upon coexposure to UV light (300-400 nm). In this paper, a comprehensive review of the literature pertaining to the toxicological interaction of PAHs and UV light in aquatic and terrestrial organisms is presented. It is concluded that the acute phototoxic effects of PAHs should be considered when conducting environmental risk assessments; however, more research needs to be conducted to address the lack of data on the enhancement of UV-induced carcinogenesis by PAH compounds.

UV-A coexposure enhances the toxicity of aromatic hydrocarbons, munitions, and metals to Photobacterium phosphoreum.

Johnson et al. (1993) showed that coexposure to UV-A between 300-400 nm enhanced the toxicity of nitrotoluenes to Photobacterium phosphoreum, a marine bioluminescent bacteria used in the Microtox test (Microbics Inc.). This paper reports that UV-A photoenhanced the toxicity of polynuclear aromatic hydrocarbons, other types of organic compounds, and some transition metals to P. phosphoreum. Coexposure to 400 muw/cm2 for 15 min increased the toxicity of psoralen, alpha-terthienyl, anthracene, acridine, fluoranthene, TNT, Cu2-, As3-, Ni2, and Cd2+. Phenanthrene was photoenhanced after 30 min coexposure at 400 muw/cm2-, and Mn2+ at 800 muw/cm2 after 15 min. Naphthalene was not enhanced at 800 muw/cm2 for 30 min.

Reversion of bioluminescent bacteria (Mutatox) to their luminescent state upon exposure to organic compounds, munitions, and metal salts.

Mutatox is a new genotoxicity bioassay which uses as the endpoint the bioluminescence produced on reversion of a dark strain of the marine bacterium Vibrio fischeri +/- S9. Reversion can occur by several mechanisms, including base substitution, frame-shift, SOS induction, and DNA intercalation. For screening, Mutatox provides many advantages over the Salmonella typhimurium (Ames) assay: it requires minimal sterility, employs a shorter incubation period, and does not require culture maintenance. Eighteen organic chemicals (phenol, polynuclear aromatic hydrocarbons, nitrotoluenes, others), Na3PO4, and 4 genotoxic metals (Cu2+, Ni2+, As3+, Cd2+) were tested. Most of the organic compounds positive in S. typhimurium assays were positive in Mutatox. None of the metals was genotoxic in V. fischeri, possibly due to poor uptake from the saline medium.