Tracking trash to treasure: in situ monitoring of single microbial cell oil biosynthesis from waste cooking oil using Raman spectroscopy and imaging.

Waste cooking oil is a major pollutant that contaminates terrestrial and aquatic bodies which is generated from household kitchens and eateries. The bioremediation of waste cooking oil (WCO) into microbial oil, also known as single microbial cell oil (SMCO), can be accomplished by oleaginous microbes. Conventional methods excel in SMCO analysis but lack efficacy for in situ or lysis-free monitoring of nascent SMCO synthesis and turnover. To bridge this knowledge gap, this study shows the applicability of Raman reverse stable isotope probing (RrSIP) in monitoring time-dependent nascent SMCO synthesis and assimilation in Yarrowia lipolytica, an oleaginous yeast grown in hydrophilic (glucose) as well as hydrophobic carbon sources (cooking oil and waste cooking oil). This study also combines the RrSIP approach with Raman imaging for temporal visualization of the distribution and turnover dynamics of the SMCO pool in a single cell. Our finding provides a unique perspective utilizing optical spectroscopy methods for lysis-free SMCO analysis and holds potential for prospective utility as an adjunct tool in bioprocess and biofuel industries.

Where the rubber meets the road: Emerging environmental impacts of tire wear particles and their chemical cocktails.

About 3 billion new tires are produced each year and about 800 million tires become waste annually. Global dependence upon tires produced from natural rubber and petroleum-based compounds represents a persistent and complex environmental problem with only partial and often-times, ineffective solutions. Tire emissions may be in the form of whole tires, tire particles, and chemical compounds, each of which is transported through various atmospheric, terrestrial, and aquatic routes in the natural and built environments. Production and use of tires generates multiple heavy metals, plastics, PAH's, and other compounds that can be toxic alone or as chemical cocktails. Used tires require storage space, are energy intensive to recycle, and generally have few post-wear uses that are not also potential sources of pollutants (e.g., crumb rubber, pavements, burning). Tire particles emitted during use are a major component of microplastics in urban runoff and a source of unique and highly potent toxic substances. Thus, tires represent a ubiquitous and complex pollutant that requires a comprehensive examination to develop effective management and remediation. We approach the issue of tire pollution holistically by examining the life cycle of tires across production, emissions, recycling, and disposal. In this paper, we synthesize recent research and data about the environmental and human health risks associated with the production, use, and disposal of tires and discuss gaps in our knowledge about fate and transport, as well as the toxicology of tire particles and chemical leachates. We examine potential management and remediation approaches for addressing exposure risks across the life cycle of tires. We consider tires as pollutants across three levels: tires in their whole state, as particulates, and as a mixture of chemical cocktails. Finally, we discuss information gaps in our understanding of tires as a pollutant and outline key questions to improve our knowledge and ability to manage and remediate tire pollution.

Toxicity of Tire Rubber Microplastics to Freshwater Sediment Organisms.

High emission of tire rubber particles to the surrounding environment is an inevitable consequence of the current habits of transportation. Although most of the emissions stay within a close range of the sources, it has been proven that the smallest particles can be transported to remote locations through the atmosphere, including inland water bodies. It has been estimated that a relevant portion of the global emissions of tire rubber particles reach surface waters, but effects on aquatic life in the receiving water bodies are not completely understood. In the present study, we used the freshwater sediment dwellers Lumbriculus variegatus and Chironomus riparius to examine the toxicity of tire rubber particles at environmentally relevant concentrations, using different types of sediment and two particle sizes of tire rubber. Overall, the experiments were unable to discern any effects on the growth, survival or reproduction of the two animals tested. Significant differences were found among the animals dwelling on different sediments, but the effects were not attributable to the presence of tire rubber particles. This study provides important information regarding the lack of effect of tire rubber particles in laboratory experiments with model sediment dwellers and opens more questions about the potential effects of tire rubber particles in the real environment with longer durations and varying environmental factors. The influence of other factors such as the leaching of additives in the overall toxicity of tire rubber particles should be also considered.

Phenotypic Plasticity of Common Wasps in an Industrially Polluted Environment in Southwestern Finland.

Insects vary in the degree of their adaptability to environmental contamination. Determining the responses with phenotypic plasticity in ecologically important species in polluted environments will ease further conservation and control actions. Here, we investigated morphological characteristics such as body size, body mass, and color of the common wasp Vespula vulgaris in an industrially polluted environment, considering different levels of metal pollution, and we studied the localization of contaminants in the guts of wasps. We revealed some differences in morphological characteristics and melanization of wasps collected in habitats with high, moderate, and low levels of pollution. The results indicated that V. vulgaris from highly polluted environments had reduced melanin pigmentation on the face but increased melanin pigmentation on the 2nd tergite of the abdomen. In addition, with transmission electron microscopy (TEM) and energy dispersive X-ray analysis (EDX), we found metal particles from the midgut of wasps originating from the polluted environment. Most of the particles were encapsulated with melanin pigment. This finding confirmed that in wasps, ingested metal particles are accumulated in guts and covered by melanin layers. Our data suggest that wasps can tolerate metal contamination but respond phenotypically with modification of their size, coloration, and probably with the directions of the melanin investments (immunity or coloration). Thus, in industrially polluted areas, wasps might probably survive by engaging phenotypic plasticity with no significant or visible impact on the population.

Altered gene expression in Chironomus riparius (insecta) in response to tire rubber and polystyrene microplastics.

The extent until which plastics are present in our surrounding environment completely exceeds our expectations. Plastic materials in the form of microplastics have been found in terrestrial, freshwater and marine environments and are transported through the atmosphere even to remote locations. However, we are still far from understanding the effects that they may have caused and are causing to biota. In the present study, we investigated the alterations in the expression of twelve genes in the aquatic insect Chironomus riparius after 36 h exposures to polystyrene and tire rubber microplastics at nominal concentrations of 1 and 10 mg L-1. The results indicated that several genes encoding for heat shock proteins (hsp90, Glycoprotein 93 (Gp93), hsc70, hsp60, hsp40, and the small HSP hsp17) were overexpressed respect to the control. In addition, the genes coding for manganese superoxide dismutase (SOD Mn, related to alleviation of oxidative stress) and for the FK506-binding protein of 39 kDa. (FKBP39, related to development and pupation) showed altered expression. Most of the alterations on gene expression level occurred at a concentration of 10 mg L-1 of tire rubber microplastics, although specific modifications arose at other concentrations of both rubber and polystyrene. On the contrary, one hsp gene (hsp10) and genes related to biotransformation and detoxification (Cyp9f2, Cyp12a2, and ABCB6) did not alter their expression in any of the treatments. Overall, the results of the gene expression indicated that microplastics (especially tire rubber) or their additives caused cellular stress that led to some alterations in the normal gene expression but did not cause any mortality after 36 h. These results highlight the need for more studies that describe the alterations caused by microplastics at the molecular level. Additionally, it opens questions about the effects caused to aquatic fauna in environmental realistic situations, especially in hot spots of microplastic contamination (e.g., tire rubber released in storm water runoff discharge points).

Is developmental instability in chironomids a sensitive endpoint for testing uranium mine-affected sediments?

There is increasing interest in effects of radionuclides on non-human species, but methods for studying such effects are not well developed. The aims of the current study were to investigate the effects of uranium mine-affected sediments on non-biting midge Chironomus riparius and to compare sensitivity of different endpoints. The midge larvae were exposed in controlled laboratory conditions to sediments from two ponds downstream from an abandoned uranium mine and a reference pond not receiving water from the mining site. Quartz sand was used as an additional control. Developmental effects were assessed by evaluating emergence of adult midges, body mass, and fluctuating asymmetry (FA) in the length of wing upper vein. FA has been suggested to be a sensitive indicator of developmental instability, but the results of previous studies are inconsistent. In the present study, no difference in FA was observed between the treatment groups, but time to emergence was significantly delayed in the contaminated sediments. The approach used in this study (laboratory experiments with sediments from a contaminated site) avoids confounding due to uncontrolled environmental variables and adaptation to long-term contamination, which may mask effects on natural populations. Using this approach, we found no effects on FA of wing length. Time to emergence, in contrast, was found to be a more sensitive endpoint.

Inhibition of pyrene biotransformation by piperonyl butoxide and identification of two pyrene derivatives in Lumbriculus variegatus (Oligochaeta).

Using the freshwater annelid Lumbriculus variegatus (Oligochaeta), the presence of cytochrome P450 (CYP) isozymes was investigated by analyzing metabolites of the polycyclic aromatic hydrocarbon (PAH) pyrene in treatments with and without the CYP inhibitor piperonyl butoxide (PBO). The results show a low biotransformation capability of L. variegatus (7% of total pyrene body burden as metabolites at 168 h). Addition of PBO resulted in a significant reduction of metabolites, suggesting the presence of a CYP in L. variegatus. Besides 1-hydroxypyrene, three peaks representing unknown metabolites were detected in LC-FLD (liquid chromatography with fluorescence detection) chromatograms of L. variegatus. Deconjugations showed that sulfonation and glucosidation are involved in the formation of these unknowns. Further studies with the time of flight mass analyzer provided the identification of the glucose-sulfate conjugate of 1-hydroxypyrene. The same metabolites were detected in the solvent-nonextractable fraction by incubation of the tissue residues with proteinase K, suggesting that part of these metabolites are bound to proteins. Overall, the slow biotransformation of pyrene by L. variegatus (involving CYP) supports the use of this species in standard bioaccumulation tests; however, the tissue-bound metabolite fraction described in the current study deserves further investigation for its toxicity and availability to upper trophic levels through diet.

Spanish population of Gremmeniella abietina is genetically unique but related to type A in Europe.

Genetic structure of the European Gremmeniella abietina var. abietina was analyzed in this study. Ninety-two Spanish isolates, six Swiss isolates of Alpine biotype, 76 Finnish isolates of biotype A and 54 Finnish and seven Russian isolates of biotype B were collected. Genetic variation of different populations was analyzed using sequence analysis of specifically amplified markers GAAA1000, GAAA800 and ACA900. Variation in the GAAA1000 marker was significant, and composed of 33 alleles divided into the following four studied populations: five alleles in the Alpine type, 12 in biotype B, 16 in biotype A and two in the Spanish population. Based on variation in GAAA1000 marker, a subset of isolates were further analyzed using GAAA800 and ACA900 sequences, which showed lower overall genetic variability, and no variation among the Spanish population. Genetic differentiation analysis revealed a high genetic differentiation among populations. Finally, clustering analysis of GAAA1000 sequences showed that the Spanish isolates clearly separated from the rest of the biotypes, whereas the Alpine type was closely related to the B type. However, one of the A-type isolates had an identical GAAA1000 allele with the prevailing allele among Spanish isolates. Altogether, our data suggest that the Spanish population is genetically highly differentiated from any other G. abietina population in Europe with a probable A-type origin.