AMR Threat Perception Assessment of Heterotrophic Bacteria From Shrimp Aquaculture Through Epidemiological Cut off Values.

BACKGROUND: Emergence and dissemination of antibiotic resistance is one of the major risks associated with the rampant usage of antibiotics in food-producing animals including aquaculture. OBJECTIVE: To determine Epidemiological Cut-OFF (ECOFF) values of heterotrophic bacterial populations from shrimp culture environments against five different antibiotics. METHODS: In this present study, bacterial samples were isolated from Penaeus vannamei culture environment in different locations of Andhra Pradesh, which is the aquaculture hub of India. The bacterial isolates were assessed for antibiotic resistance towards five antibiotics belonging to different classes (oxytetracycline, chloramphenicol, erythromycin, ciprofloxacin, and co-trimoxazole) by the disc diffusion method. Determination of Epidemiological Cut-OFF (ECOFF) values and analysis by employing normalized resistance interpretation (NRI) was carried out. RESULTS: The most dominant bacterial populations from shrimp culture were Vibrio spp. (pathogenic bacteria) followed by Bacillus spp. (probiotic bacteria). The bacterial isolates showed highest resistance towards oxytetracycline (overall 23.38%) and in location L6 (59.4%) followed by co-trimoxazole (31.1%). ECOFF values calculated by employing NRI showed that the disc diffusion data were distributed in a normalized manner. The maximum ECOFF value was obtained for ciprofloxacin (23.32 mm), while the minimum value was observed for oxytetracycline (9.05 mm). The antibiotic resistant phenotypes showed that the majority of the heterotrophic bacterial isolates (>60%) belonged to the non-wild type phenotype and primarily towards oxytetracycline (90%). CONCLUSION: The presence of non-wild antibiotic-resistant phenotypes of heterotrophic bacterial populations (which include not only pathogenic bacteria but also probiotic bacteria) indicates that shrimp culture ponds may be a reservoir for drug-resistant bacteria and there is a greater risk associated with transmission of resistant genes across bacterial flora. HIGHLIGHTS: NRI analysis of antibiotic disc diffusion data of heterotrophic bacterial populations in shrimp aquaculture environments revealed that majority of them belonged to non-wild type (90%) paticularly to oxytetracycline in comparison to other studied antibiotics (chloramphenicol, erythromycin, ciprofloxacin and co-trimoxazole).

Chlorination and bromination of 1,3-diphenylguanidine and 1,3-di-o-tolylguanidine: Kinetics, transformation products and toxicity assessment.

This works investigates the chlorination and bromination of two rubber and polymer related chemicals, which have emerged as relevant water contaminants, i.e. 1,3-di-o-tolylguanidine (DTG) and 1,3-diphenylguanidine (DPG). Kinetic constants at different pH values were obtained and modelled, taking into account the pKa values of DTG/DPG and HClO, showing that the maximum reaction rate (kapp > 104 M-1 s-1) is obtained at pH values 8.8 for DPG and 9.1 for DTG. Bromination is also very fast, although unlike chlorination, deviation from the model was observed at neutral pH, which was attributed to formation of metastable transformation product (TP). A total of 35 TPs, corresponding to halogenation, hydroxylation, formation of monophenylguanidine derivatives and cyclization reactions, were tentatively identified. Furthermore it was found that chloroform can be formed up to a 25% molar yield, while dichloroacetonitrile was formed into less than a 3% yield. Several ecotoxicological endpoints were predicted by quantitative structure-activity relationship models (QSAR) for the TPs, some of which were predicted to be more toxic than DPG/DTG. Also a chlorinated solution investigated by a Vibrio Fisheri acute toxicity test, confirmed that toxicity increases with chlorination.

Efficacy assessment of commercially available natural products and antibiotics, commonly used for mitigation of pathogenic Vibrio outbreaks in Ecuadorian Penaeus (Litopenaeus) vannamei hatcheries.

Bacterial diseases cause high mortality in Penaeus (Litopenaeus) vannamei postlarvae. Therefore, appropriate application of efficient therapeutic products is of vital importance for disease control. This study evaluated through in vitro analyses the antimicrobial effectiveness of commercial therapeutic products used for P. vannamei bacterial diseases and antibiotics against pathogenic Vibrio strains circulating in Ecuadorian hatcheries. Twenty strains were isolated from 31 larvae samples with high bacterial counts from 10 hatcheries collected during mortality events. The strains virulence was verified through challenge tests with Artemia franciscana nauplii and P. vannamei postlarvae. Through 16S rRNA sequence analysis, strains showed a great similarity to the Vibrio sequences reported as pathogens, with 95% belonging to the Harveyi clade. Through antibiograms and minimal inhibitory concentration (MIC) in vitro tests we found that furazolidone, ciprofloxacin, chloramphenicol, norfloxacin, nalidixic acid, florfenicol, fosfomycin and enrofloxacin inhibited the growth of all or most of the strains. Less efficient antibiotics were penicillin, oxytetracycline and tetracycline. A multiple antibiotic resistance (MAR) index of 0.23 showed some level of resistance to antibiotics, with two MAR prevalent patterns (Penicillin-Oxytetracycline and Penicillin-Oxytetracycline-Tetracycline). From a total of 16 natural products (five probiotics, nine organic acids and two essential oils), only three (one probiotic, one organic acid and one essential oil) were effective to control most of the strains. Shrimp producers can apply relatively simple in vitro analyses, such as those employed in this study, to help take adequate management decisions to reduce the impact of bacterial diseases and increase profit.

Ecotoxicity assessment of boreal lake sediments affected by metal mining: Sediment quality triad approach complemented with metal bioavailability and body residue studies.

There are several methods for studying metal-contaminated freshwater sediments, but more information is needed on which methods to include in ecological risk assessment. In this study, we compliment the traditional Sediment Quality Triad (SQT) approach - including information on chemistry, toxicity and ecological status - with studies on metal bioavailability and metal body residues in local organisms. We studied four mining-affected boreal lakes in Finland by conducting chemical analyses of sediment and water, toxicity tests (L. variegatus, V. fischeri, C. riparius, L. stagnalis), and analysis of benthic organism community structure. In addition, we studied the relationships between metal loading, toxicity, metal bioavailability, and metal body residues in the field-collected biota. Chemistry and benthic organism community structures show adverse effects in those lakes, where the metal concentrations are the highest. However, toxicity was connected to low sediment pH during the experiment, rather than to high metal concentrations. Toxicity was observed in 4 out of 6 toxicity tests including growth test with L. variegatus, bulk sediment test with V. fischeri, and the L. stagnalis toxicity test. The C. riparius test did not show toxicity. Metal body residues in biota were not high enough to induce adverse effects (0.1-4.1 mg Cu/kg fw, 0.01-0.3 mg Ni/kg fw, 2.9-26.7 mg Zn/kg fw and 0.01-0.7 mg As/kg fw). Chemical analyses, metal bioavailability assessment and benthic community structures survey revealed adverse effects in the sediments, where metal concentrations are highest (Lake SJ and Lake KS). Standard toxicity tests were not suitable for studying acid, sulfide-rich sediments and, therefore, benthic structure study and chemical analyses are believed to give more reliable results of the ecological status of these sediments.

Environmental hazard assessment by the Ecoscore system to discriminate PAH-polluted soils.

A bioassay battery-integrated index was applied to different soils sampled from a former coke factory, with the aim to evaluate the discriminating capacity of the Ecoscore system (ES) to assess the environmental hazard of PAH-polluted soils. Two soils from a former coke factory, polluted with polycyclic aromatic hydrocarbons (PAHs), were evaluated for their ecotoxicity to terrestrial and aquatic organisms and their genotoxicity. These soils have been already presented in a previous paper but data have been reanalyzed for the present article in an endeavor to standardize the ES. One soil was sampled in the untreated site and the second underwent a windrow treatment. While these soils had a similar total concentrations of US-EPA 16PAHs (around 3000 mg kg-1), different ecoscores were obtained when subjected to a set of solid- and liquid-phase bioassays measuring acute, chronic, and genotoxic effects. The total PAH content of the soil is not a pertinent parameter to assess soil pollution hazards contrary to the ES. ES is a robust method to classify soils according to their toxicity level. Four levels of toxicity have been defined: no (ecoscore = 0), weak (0 < ecoscore ≤33), moderate (33 < ecoscore ≤67), and strong toxicity (67 < ecoscore ≤ 100). The combination of chemical and toxicological data highlights the relationship between three-ring PAHs and acute ecotoxicity. Conversely, chronic effects of water extracts on algal growth could be explained by high molecular weight PAHs, such as five- and six-ring PAHs.

Rapid Assessment of the Toxicity of Fungal Compounds Using Luminescent Vibrio qinghaiensis sp. Q67.

Most tropical fruits after harvest are very perishable because of fungal infection. Since some pathogenic fungi can produce hazardous compounds such as mycotoxins, novel rapid and effective methods to assess those hazardous compounds are urgently needed. Herein we report that Vibrio qinghaiensis sp. Q67, a luminescent bacterium, can be used to rapidly assess the toxicities of mycotoxins and cultures from mycotoxin-producing pathogens. A good correlation (R² > 0.98) between concentrations of the mycotoxins (fumonisin B1, deoxynivalenol, zearalenone, ochratoxin A, patulin, and citrinin) and the luminous intensity of V. qinghaiensis sp. Q67 was obtained. Furthermore, significant correlations (R² > 0.96) between the amount of mycotoxin and the luminous intensity from the cultures of 10 major mycotoxin-producing pathogens were also observed. In addition, Fusarium proliferatum (half-maximal inhibitory concentration (IC50) = 17.49%) exhibited greater luminescence suppression than Fusarium semitectum (IC50 = 92.56%) or Fusarium oxysporum (IC50 = 28.61%), which was in agreement with the existing higher levels of fumonisin B1, fumonisin B2, and deoxynivalenol, which were measured by high-performance liquid chromatography-tandem mass spectrometry. These results suggest that V. qinghaiensis sp. Q67 is a promising alternative for the rapid evaluation of the toxicity of fungal mycotoxins.

Vibrio Pathogens: A Public Health Concern in Rural Water Resources in Sub-Saharan Africa.

Members of the Vibrio genus are autochthonous inhabitants of aquatic environments and play vital roles in sustaining the aquatic milieu. The genus comprises about 100 species, which are mostly of marine or freshwater origin, and their classification is frequently updated due to the continuous discovery of novel species. The main route of transmission of Vibrio pathogens to man is through drinking of contaminated water and consumption inadequately cooked aquatic food products. In sub-Saharan Africa and much of the developing world, some rural dwellers use freshwater resources such as rivers for domestic activities, bathing, and cultural and religious purposes. This review describes the impact of inadequately treated sewage effluents on the receiving freshwater resources and the associated risk to the rural dwellers that depends on the water. Vibrio infections remain a threat to public health. In the last decade, Vibrio disease outbreaks have created alertness on the personal, economic, and public health uncertainties associated with the impact of contaminated water in the aquatic environment of sub-Saharan Africa. In this review, we carried out an overview of Vibrio pathogens in rural water resources in Sub-Saharan Africa and the implication of Vibrio pathogens on public health. Continuous monitoring of Vibrio pathogens among environmental freshwater and treated effluents is expected to help reduce the risk associated with the early detection of sources of infection, and also aid our understanding of the natural ecology and evolution of Vibrio pathogens.

Toxicity evaluation of wastewater collected at different treatment stages from a pharmaceutical industrial park wastewater treatment plant.

The toxicity of water-receiving bodies, the effluent and other treatment stages in wastewater treatment plants has recently been of interest to the public due to the lack of a regulated toxicity-based index for wastewater discharge in China. This study aimed to evaluate the conventional pollution parameters and toxicities of wastewaters collected at different treatment stages from a pharmaceutical industrial park wastewater treatment plant through dehydrogenase activity (DHA) and bioluminescent bacteria (Vibrio qinghaiensis) tests. The results of an analysis of conventional parameters indicated that the total suspended solids (TSS), chemical oxygen demand (COD), total nitrogen (TN), ammonia nitrogen (NH3N), and total phosphorus (TP) were largely removed after various treatments. However, the TN, NH3N and COD still exceeded the regulated standards. The tested pharmaceutical park effluents were mainly polluted with organic pollutants and nitrogenous. The toxicity test results indicated that the toxicities could be markedly reduced after treatment, with the toxicities of two out of the six effluent samples at different treatment stages being greater than the influent toxicity. Spearman's rank correlation coefficients indicated a significantly positive correlation between the toxicity values obtained using the DHA and Vibrio qinghaiensis tests. Compared with the DHA measurement, the Vibrio qinghaiensis test was faster and more sensitive. Meanwhile, the toxicity indicators were significantly and positively correlated with the TSS, TN, TP and COD concentrations. These results may aid the understanding of the toxicity of pharmaceutical industrial park wastewaters and toxicity removal using the treatment techniques that are currently utilized in China.

Personal care product preservatives: risk assessment and mixture toxicities with an industrial wastewater.

The aquatic toxicity of eight preservatives frequently used in personal care products (PCPs) (iodopropynyl butylcarbamate, bronopol, diazolidinyl urea, benzalkonium chloride, zinc pyrithione, propylparaben, triclosan and a mixture of methylchloroisothiazolinone and methylisothiazolinone) was assessed by means of two different approaches: a battery of bioassays composed of single species tests of bacteria (Vibrio fischeri and Pseudomonas putida) and protozoa (Tetrahymena thermophila), and a whole biological community resazurin-based assay using activated sludge. The tested preservatives showed considerable toxicity in the studied bioassays, but with a marked difference in potency. In fact, all biocides except propylparaben and diazolidinyl urea had EC50 values lower than 1 mg L(-1) in at least one assay. Risk quotients for zinc pyrithione, benzalkonium chloride, iodopropynyl butylcarbamate and triclosan as well as the mixture of the studied preservatives exceeded 1, indicating a potential risk for the process performance and efficiency of municipal sewage treatment plants (STPs). These four single biocides explained more than 95% of the preservative mixture risk in all bioassays. Each individual preservative was also tested in combination with an industrial wastewater (IWW) from a cosmetics manufacturing facility. The toxicity assessment was performed on binary mixtures (preservative + IWW) and carried out using the median-effect principle, which is a special case of the concept of Concentration Addition (CA). Almost 70% of all experiments resulted in EC50 values within a factor of 2 of the values predicted by the median-effect principle (CI values between 0.5 and 2). The rest of the mixtures whose toxicity was mispredicted by CA were assessed with the alternative concept of Independent Action (IA), which showed higher predictive power for the biological community assay. Therefore, the concept used to accurately predict the toxicity of mixtures of a preservative with a complex industrial wastewater depends on degree of biological complexity.

Assessment of planctomycetes cell viability after pollutants exposure.

In this study, the growth of six different planctomycetes, a particular ubiquitous bacterial phylum, was assessed after exposure to pollutants. In addition and for comparative purposes, Pseudomonas putida, Escherichia coli and Vibrio anguillarum were tested. Each microorganism was exposed to several concentrations of 21 different pollutants. After exposure, bacteria were cultivated using the drop plate method. In general, the strains exhibited a great variation of sensitivity to pollutants in the order: V. anguillarum > planctomycetes > P. putida > E. coli. E. coli showed resistance to all pollutants tested, with the exception of phenol and sodium azide. Copper, Ridomil® (fungicide), hydrazine and phenol were the most toxic pollutants. Planctomycetes were resistant to extremely high concentrations of nitrate, nitrite and ammonium but they were the only bacteria sensitive to Previcur N® (fungicide). Sodium azide affected the growth on plates of E. coli, P. putida and V. anguillarum, but not of planctomycetes. However, this compound affected planctomycetes cell respiration but with less impact than in the aforementioned bacteria. Our results provide evidence for a diverse response of bacteria towards pollutants, which may influence the structuring of microbial communities in ecosystems under stress, and provide new insights on the ecophysiology of planctomycetes.

Reverse osmosis pretreatment method for toxicity assessment of domestic wastewater using Vibrio qinghaiensis sp.-Q67.

Luminescent bacterial test is a fast and sensitive method for acute toxicity assessment of water and wastewater. In this study, an improved toxicity testing method was developed using the freshwater luminescent bacteria Vibrio qinghaiensis sp.-Q67 that involved pretreatment of water samples with reverse osmosis (RO) to eliminate the interferences caused by nutrients in concentrated samples and to improve the reliability and sensitivity of the analysis. Because water samples contain low concentrations of several target toxic substances, rapid acute toxicity testing method that is commonly employed does not achieve enough sensitivity. The proposed RO pretreatment could effectively enrich organic and inorganic substances in water samples to enable a more effective and sensitive toxicity evaluation. The kinetic characteristics of toxicity of raw sewage and secondary effluent were evaluated based on the relative luminescence unit (RLU) curves and time-concentration-effect surfaces. It was observed that when the exposure time was prolonged to 8-h or longer, the bacteria reached the logarithmic growth stage. Hence, the stimulating effects of the coexisting ions (such as Na(+), K(+), NO3(-)) in the concentrated samples could be well eliminated. A 10-h exposure time in proposed Q67 test was found to quantitatively evaluate the toxicity of the organic and inorganic pollutants in the RO-concentrated samples.

Mutagenicity test using Vibrio harveyi in the assessment of water quality from mussel farms.

This work analyses the mutagenicity of seawater from mussel farms using the Vibrio harveyi mutagenicity test and its relationship with the accumulated pollutants and the development of gonadal neoplasia in mussels. Histological disorders identified as germinoma were observed in the gonad of Mytilus galloprovincialis during the period of study. The prevalence of this pathology is significantly correlated with certain levels of pollutants accumulated in mussels, mainly of PAHs and PCBs, whose toxic equivalents were calculated as EROD induction equivalency. The mutagenicity and toxicity of the water surrounding mussel's farms is clearly correlated with the pollutants accumulated and with the neoplasia prevalence in mussels. Such correlations are corroborated by a multivariate analysis. Our results conclude with the utility of V. harveyi test as an optimal and rapid method in the monitoring of the quality of the water from mussel farms and as a tool to control the risks of pollution on mussel production and its safety for human food.

How toxic is toxic? A proposal for wastewater toxicity hazard assessment.

Wastewater management receives a great deal of attention with various methods being proposed for discharge hazard estimation via ecotoxicological results. Policy-makers, stakeholders and the general public do not generally possess an adequate level of understanding on this matter, so it is rather hard to answer the question "How toxic is toxic?". The setting up and development of species-specific toxicity scores and a final wastewater toxicity index could avoid misinterpretations and confusion about toxicity data and different endpoints used and thus help wastewater classification and the management actions to be undertaken. Five-class toxicity scores were developed considering saltwater species. Toxicity scores outputs were then considered for a final index definition. This approach for wastewater assessment could be a suitable way to proceed in order to achieve environmental protection of water bodies, both fresh and saltwater, in accordance with the (near-)zero emission approach and the precautionary principle.

The use of the Vibrio harveyi luminescence mutagenicity assay as a rapid test for preliminary assessment of mutagenic pollution of marine sediments.

Mutagenic pollution of the natural environment is currently one of the most serious environmental problems. It includes the pollution of marine sediments. Therefore, rapid detection of the presence of mutagens is an important issue. Recently, we have developed a novel microbiological assay for rapid assessment of mutagenicity of samples from the natural environment. This assay is based on bioluminescence of a mutant Vibrio harveyi strain, and was shown to be useful in testing samples of marine water and plant tissues. Here we demonstrate the usefulness of this assay in preliminary assessment of mutagenic pollution of marine sediments. Mutagenicity of environmental samples taken from the Baltic Sea, is documented and compared here with a commercially available standard sediment sample (IAEA 383), which contains known amounts of mutagenic compounds. The whole procedure, from obtaining a sample in the laboratory to getting final results, is very short (less than 4 h).

Photoelectrocatalytic oxidation of remazol turquoise blue and toxicological assessment of its oxidation products.

The ability of photoelectrocatalytic oxidation to degrade the commercially important copper-phtalocyanine dye, remazol turquoise blue 15 (RTB) was investigated. The best experimental condition was optimized, evaluating the performance of Ti/TiO2 thin-film electrodes prepared by sol-gel method in the decolourization of 32 mg L(-1) RTB dye in 0.5 mol L(-1) Na2SO4 pH 8 and applied potential of +1.5 V versus SCE under UV irradiation. Spectrophotometric measurements, high performance liquid chromatography, dissolved organic carbon (TOC) evaluation and stripping analysis of yielding solution obtained after 3 h of photoelectrolysis leads to 100% of absorbance removal from wavelength of 250-800 nm, 79.6% of TOC reduction and the releasing of up to 54.6% dye-bound copper (0.85 mg L(-1)) into the solution. Both, original and oxidized dye solution did not presented mutagenic activity with the strains TA98 and TA100 of Salmonella in the presence and absence of S9 mix at the tested doses. Nevertheless, the yielding photoelectrocatalytic oxidized solution showed an increase in the acute toxicity for Vibrio fischeri bacteria, explained by copper liberation during treatment.

Toxicity assessment of contaminated soils from an antitank firing range.

Explosives are released into the environment at production and processing facilities, as well as through field use. These compounds may be toxic at relatively low concentrations to a number of ecological receptors. A toxicity assessment was carried out on soils from an explosive-contaminated site at a Canadian Forces Area Training Center. Toxicity studies on soil organisms using endpoints such as microbial processes (potential nitrification activity, dehydrogenase activity, substrate-induced respiration, basal respiration), plant seedling and growth (Lactuca sativa and Hordeum vulgare), and earthworm (Eisenia andrei) growth and reproduction were carried out. Results showed that 1,3,5,7-tetranitro-1,3,5,7-tetrazacyclooctane (HMX) was the principal polynitro-organic compound measured in soils. Soils from the contaminated site decreased microbial processes and earthworm reproduction; whereas plant growth was not significantly reduced. Toxicity to aquatic organisms and genotoxicity were also assessed on soil elutriates using Microtox (Vibrio fischeri), growth inhibition of algae (Selenastrum capricornutum), and SOS Chromotest (Escherichia coli). Results indicated that soil elutriates were generally not toxic to bacteria (Microtox) and algae. However, genotoxicity was found in a number of soil elutriate samples. Thus, the explosive-contaminated soils from the antitank firing range may represent a hazard for the soil organisms. Nevertheless, the global toxicity might have partially resulted from HMX as well as from other (not identified) contaminants such as heavy metals.

Ozonation of an azo dye C.I. Remazol Black 5 and toxicological assessment of its oxidation products.

The effect of ozonation (20.5 mgl(-1)) on the degradation processes of an azo dye, Remazol Black 5 (RB5; CI) was studied. Conventional parameters such as chemical oxygen demand (COD), total organic carbon (TOC), pH, conductivity, colour removal, biodegradability (BOD(5/28)), and toxic potential of the dye and its degradation products were monitored during the process. The results obtained indicated that ozonation is a highly effective way to remove the colour of a corresponding dye solution. However, a considerable organic load still remained as indicated by high COD and TOC residues. The COD, TOC reductions were about 40% and 25% for 6 h ozonation of 2 gl(-1) RB5 aqueous solution. During the ozonation process the rapid decrease of pH and the sharp increase of conductivity indicated the formation of acidic by-products and small fragments and ions which were identified by high performance ion chromatography. The BOD28 data revealed that first by-products after partial ozonation (10-150 min) of RB5 were more biodegradable than the parent compound and ozonation can enhance the biodegradability of azo dyes. During the first 150 min of total 360 min of oxidation, the formation of first by-products with high toxic potential took place as it could be confirmed by two acute toxicity-screening tests, the bioluminescence test (Vibrio fischerii) and the neutral red cytotoxicity assay (rat hepatoma cells). The significant enhancement of microbial biodegradability after long-term ozonation could also be seen as a decrease of toxic intermediates in correlation with the ozonation time as indicated in BOD28 biological degradation test results.

Assessment of the influence of use on ecotoxicological characteristics of synthetic ester lubricants.

Synthetic ester lubricants need optimisation about their technical and their ecotoxicological characteristics. To determine the ecotoxicological potential the required examinations can be based on the procedure for a risk assessment of chemicals. At present risk classification of lubricant oils is carried out with new oil fluids that are normally prepared before application in aqueous bioassays. In order to improve the ecotoxicological characteristics of some lubricant oils, the quality of the preparation method has been optimised. The resulting preparation protocol leads to aqueous extracts of the oil fluids that can be tested using biological assays. The extent of the changes of the chemical composition caused by the use as well as the ecotoxicological effects caused by additives have to be taken into consideration. For this reason various used lubricants are tested in addition to new oil fluids. In this work various lubricant samples were examined with standardised bacterial growth assays with Vibrio fischeri and Pseudomonas putida, luminescence inhibition assay with V. fischeri, survival assay with Daphnia magna and algal growth inhibition assay with Scenedesmus subspicatus. The chemical characterisation of the aqueous extracts included the determination of pH, conductivity, heavy metals, the content of dissolved organic carbon, inorganic anions and the content of phosphorus. The results emphasize the thesis that environmentally acceptable lubricants can undergo a change of their ecotoxicological potential during the use. Some of the substances that are normally added to base fluids in order to enhance the applicability of the oils may possess a high toxicological potential.

Assessment of dental material degradation product toxicity using a bioluminescent bacterial assay.

OBJECTIVES: This study examined dental material degradation product toxicity using the Microtox bacterial bioluminescence assay as well as the effects on toxicity of selective leaching, chelation with protein, the physical form of the products, and synergistic/antagonistic interactions among released ions. METHODS: Polarization was used to produce ionically dissolved (ID) and precipitated corrosion products from Litecast B alloy specimens, which were then chemically analyzed to determine their composition and to identify metal valence states. Corrosion product toxicity, as well as that of the individual alloying elements, alone and in the presence of mucin, was analyzed using Microtox. A mathematical approach identified synergistic/antagonistic interactions and determined element contribution to product toxicity. The mechanism by which the Microtox test bacterium interacts with solid products was explored. The toxicity of methyl methacrylate (MMA) monomer was also examined. RESULTS: Precipitated corrosion products were found to be more toxic than ID products. The metals in the precipitate have been shown to be available to the test bacterium. Be and Ni were the most toxic elements in the products and contributed significantly to their toxicity. Synergistic and slightly antagonistic interactions were observed in the ID and precipitated products, respectively. Mucin decreased toxicity of all elements except Be. MMA monomer toxicity was found to be low compared to metal toxicity. SIGNIFICANCE: Microtox is useful for evaluating dental degradation product biocompatibility and has significant promise for use in other types of studies, such as determining the effectiveness of antimicrobial treatments.