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Life cycle assessment part 1: framework, goal and scope definition, inventory analysis, and applications.

Sustainable development requires methods and tools to measure and compare the environmental impacts of human activities for the provision of goods and services (both of which are summarized under the term "products"). Environmental impacts include those from emissions into the environment and through the consumption of resources, as well as other interventions (e.g., land use) associated with providing products that occur when extracting resources, producing materials, manufacturing the products, during consumption/use, and at the products' end-of-life (collection/sorting, reuse, recycling, waste disposal). These emissions and consumptions contribute to a wide range of impacts, such as climate change, stratospheric ozone depletion, tropospheric ozone (smog) creation, eutrophication, acidification, toxicological stress on human health and ecosystems, the depletion of resources, water use, land use, and noise-among others. A clear need, therefore, exists to be proactive and to provide complimentary insights, apart from current regulatory practices, to help reduce such impacts. Practitioners and researchers from many domains come together in life cycle assessment (LCA) to calculate indicators of the aforementioned potential environmental impacts that are linked to products-supporting the identification of opportunities for pollution prevention and reductions in resource consumption while taking the entire product life cycle into consideration. This paper, part 1 in a series of two, introduces the LCA framework and procedure, outlines how to define and model a product's life cycle, and provides an overview of available methods and tools for tabulating and compiling associated emissions and resource consumption data in a life cycle inventory (LCI). It also discusses the application of LCA in industry and policy making. The second paper, by Pennington et al. (Environ. Int. 2003, in press), highlights the key features, summarises available approaches, and outlines the key challenges of assessing the aforementioned inventory data in terms of contributions to environmental impacts (life cycle impact assessment, LCIA).

Conservation of Natural Resources↗

The human papillomavirus type 16 E7 oncogene is required for the productive stage of the viral life cycle.

The production of the human papillomavirus type 16 (HPV-16) is intimately tied to the differentiation of the host epithelium that it infects. Infection occurs in the basal layer of the epithelium at a site of wounding, where the virus utilizes the host DNA replication machinery to establish itself as a low-copy-number episome. The productive stage of the HPV-16 life cycle occurs in the postmitotic suprabasal layers of the epithelium, where the virus amplifies its DNA to high copy number, synthesizes the capsid proteins (L1 and L2), encapsidates the HPV-16 genome, and releases virion particles as the upper layer of the epithelium is shed. Papillomaviruses are hypothesized to possess a mechanism to overcome the block in DNA synthesis that occurs in the differentiated epithelial cells, and the HPV-16 E7 oncoprotein has been suggested to play a role in this process. To determine whether E7 plays a role in the HPV-16 life cycle, an E7-deficient HPV-16 genome was created by inserting a translational termination linker (TTL) in the E7 gene of the full HPV-16 genome. This DNA was transfected into an immortalized human foreskin keratinocyte cell line shown previously to support the HPV-16 life cycle, and stable cell lines were obtained that harbored the E7-deficient HPV-16 genome episomally, the state of the genome found in normal infections. By culturing these cells under conditions which promote the differentiation of epithelial cells, we found E7 to be necessary for the productive stage of the HPV-16 life cycle. HPV-16 lacking E7 failed to amplify its DNA and expressed reduced amounts of the capsid protein L1, which is required for virus production. E7 appears to create a favorable environment for HPV-16 DNA synthesis by perturbing the keratinocyte differentiation program and inducing the host DNA replication machinery. These data demonstrate that E7 plays an essential role in the papillomavirus life cycle.

Apoptosis↗

Multicellular life cycle of magnetotactic prokaryotes.

Most multicellular organisms, prokaryotes as well as animals, plants, and algae have a unicellular stage in their life cycle. Here, we describe an uncultured prokaryotic magnetotactic multicellular organism that reproduces by binary fission. It is multicellular in all the stages of its life cycle, and during most of the life cycle the cells organize into a hollow sphere formed by a functionally coordinated and polarized single-cell layer that grows by increasing the cell size. Subsequently, all the cells divide synchronously; the organism becomes elliptical, and separates into two equal spheres with a torsional movement in the equatorial plane. Unicellular bacteria similar to the cells that compose these organisms have not been found. Molecular biology analysis showed that all the organisms studied belong to a single genetic population phylogenetically related to many-celled magnetotactic prokaryotes in the delta sub-group of the proteobacteria. This appears to be the first report of a multicellular prokaryotic organism that proliferates by dividing into two equal multicellular organisms each similar to the parent one.

Brazil↗

The minor capsid protein L2 contributes to two steps in the human papillomavirus type 31 life cycle.

Prior studies, which have relied upon the use of pseudovirions generated in heterologous cell types, have led to sometimes conflicting conclusions regarding the role of the minor capsid protein of papillomaviruses, L2, in the viral life cycle. In this study we carry out analyses with true virus particles assembled in the natural host cell to assess L2's role in the viral infectious life cycle. For these studies we used the organotypic (raft) culture system to recapitulate the full viral life cycle of the high-risk human papillomavirus HPV31, which was either wild type or mutant for L2. After transfection, the L2 mutant HPV31 genome was able to establish itself as a nuclear plasmid in proliferating populations of poorly differentiated (basal-like) human keratinocytes and to amplify its genome to high copy number, support late viral gene expression, and cause formation of virus particles in human keratinocytes that had been induced to undergo terminal differentiation. These results indicate that aspects of both the nonproductive and productive phases of the viral life cycle occur normally in the absence of functional L2. However, upon the analysis of the virus particles generated, we found an approximate 10-fold reduction in the amount of viral DNA encapsidated into L2-deficient virions. Furthermore, there was an over-100-fold reduction in the infectivity of L2-deficient virus. Because the latter deficiency cannot be accounted for solely by the 10-fold decrease in encapsidation, we conclude that L2 contributes to at least two steps in the production of infectious virus.

Capsid Proteins↗

Induction of productive human papillomavirus type 11 life cycle in epithelial cells grown in organotypic raft cultures.

The study of the human papillomavirus (HPV) life cycle was hampered for more than 50 years by the lack of a conventional cell culture system for propagating HPV. Considerable progress has been made in the production of several HPV types using either organotypic rafts or human epithelial xenografts in immunocompromised mice. In this study, we demonstrated episomal maintenance of HPV-11 DNA in N-Tert cells. HPV-11 episomal DNA containing cell populations grown in raft culture showed induction of the productive viral life cycle. HPV-11 DNA amplification and viral capsid antigen synthesis were detected in differentiated layers of epithelia. The viruses generated were able to infect keratinocytes in vitro, which indicate that viruses generated were infectious. The demonstration of the productive HPV-11 life cycle in raft culture from cloned HPV-11 DNA will facilitate genetic analyses of viral gene functions that was not possible using the human xenograft athymic mouse model.

Animals↗

The role of life cycle and migration in selection for variance in offspring number.

For two genotypes that have the same mean number of offspring but differ in the variance in offspring number, naturalselection will favor the genotype with lower variance. In such cases, the average growth rate is not sufficient as a measure of fitness or as a predictor of fixation probability. However, the effect of variance in offspring number on the fixationprobability of mutant strategies has been calculated under several scenarios with the general conclusion that variance in offspring number reduces fitness in proportion to the inverse of the population size [Gillespie, J., Genetics 76:601-606, 1974; Proulx, S.R., Theor. Popul. Biol. 58:33-47, 2000]. This relationship becomes more complicated under a metapopulation scenario where the "effective" population size depends on migration rate, population structure, and lifecycle. It is shown that in a life cycle where reproduction and migration (the birth-migration-regulation life cycle, or BMR)occur prior to density regulation within every deme, the fitness of a strategy depends on migration rate. When migration rates are near zero, the fitness of the strategy is determined by the size of individual demes, so that the strategy favoredin small populations tends to be fixed. As migration rate increases and approaches panmixis between demes, the fitness ofa reproductive strategy approaches what its value would be in a single, panmictic deme with a population size correspondingtothe census size of the metapopulation. Interestingly, when the life cycle is characterized by having density regulation in each deme prior to migration (the BRM life cycle) the fixation probability of a strategy is independent of migration rate. These results are found to be qualitatively consistent with the individual-based simulation results in Shpak [Theor. Biosci.124:65-85, 2005].

Alleles↗

Life cycle and morphology of Metagonimus miyatai (Digenea: Heterophyidae) from Nagano, Japan.

The life cycle of Metagonimus miyatai Saito, Chai, Kim, Lee and Rim, 1997 (Digenea: Heterophyidae) was studied in the Hiroi River at Kotobuki, Iiyama City, Nagano Prefecture, central Japan. Daughter rediae and cercariae were found in Semisulcospira libertina (Gould) and S. dolorosa (Gould) (Gastropoda: Pleuroceridae). Metacercariae were found encysted in Phoxinus lagowskii steindachneri Sauvage (Osteichthyes: Cyprinidae). Cercariae were experimentally exposed to P. lagowskii steindachneri, and metacercariae were obtained from the fish. Metacercariae of natural and experimental infections were experimentally fed to respective golden hamsters, and gravid adults identifiable as M. miyatai were recovered from the small intestine of the golden hamsters. The daughter redia, cercaria, metacercariae and adults are described. The life cycle of M. miyatai is discussed.

Animals↗

Infraciliature, morphogenesis and life cycle of Endosphaera terebrans (Suctoria, Tokophridae).

The morphology, infraciliature, and life cycle of Endosphaera terebrans, a suctorian endocommensal of peritrichs, have been studied with the aid of silver impregnation. The life cycle of Endosphaera terebrans begins with infection of the host cell by a small larva. The swarmer has a pointed needle-like cellular projection and two rings of cilia. The swarmer penetrates the the peritrich, loses the cilia, and then matures into an adult. The infraciliature of the adult form has four rows of barren kinetosomes that lack kinetodesmal fibers. By endogenous budding, a migratory larva is produced that leaves the host cell through the peristomial disc and that can infect other peritrichs.

Animals↗

Antigenic differences between the life cycle stages of Cooperia oncophora.

The antigenic differences among the life cycle stages of Cooperia oncophora were studied by SDS-gel electrophoresis and Western blotting. Sodium dodecylsulphate polyacrylamide gel electrophoresis of the different life cycle stages of C oncophora revealed a complex protein pattern with a decreasing number of protein bands towards the adult stages. Several bands of the fourth stage larvae were in common with both the third stage and the adult nematode. Western blotting with sera from C oncophora monoinfected calves showed that the antigens of the fourth stage larvae were recognised predominantly and the presence of stage specific antigens in all stages. Strong cross reactivity was demonstrated when serum from Ostertagia ostertagi infected calves was used.

Animals↗

Comparison of a partial life-cycle bioassay in artificial streams to a standard beaker bioassay to assess effects of metal mine effluent on Chironomus tentans.

A novel, partial life-cycle bioassay using Chironomus tentans in artificial streams was developed for evaluating the effects of metal mine effluent. The utility of this bioassay was compared to an existing beaker life-cycle bioassay under laboratory conditions. Chironomus tentans larvae were exposed to 45% (v/v) treated metal mine effluent from day 11 through hatching of the second generation. Response patterns were consistent between the two bioassays for hatching success and time to emergence but inconsistent for other endpoints. Significant effects of effluent were obtained for growth, survival, number of adults emerged, and number of eggs per egg case in the artificial stream bioassay but not in the beaker bioassay. Conversely, significant effects on sex ratio and number of egg cases per female were observed in the beaker bioassay but not in the artificial stream bioassay. These differences are believed to be a consequence of the number of organisms per replicate used in each bioassay, which results in a difference in statistical power. As a result, higher coefficients of variation and effects sizes were observed in the beaker bioassay relative to the artificial stream bioassay for almost all endpoints. These results provide evidence that the bioassay in artificial streams can be an effective tool for evaluating the effects of metal mine effluent on life-cycle endpoints in C. tentans.

Animals↗

Development of the endoplasmic reticulum, mitochondrion and apicoplast during the asexual life cycle of Plasmodium falciparum.

Plasmodium parasites are unicellular eukaryotes that undergo a series of remarkable morphological transformations during the course of a multistage life cycle spanning two hosts (mosquito and human). Relatively little is known about the dynamics of cellular organelles throughout the course of these transformations. Here we describe the morphology of three organelles (endoplasmic reticulum, apicoplast and mitochondrion) through the human blood stages of the parasite life cycle using fluorescent reporter proteins fused to organelle targeting sequences. The endoplasmic reticulum begins as a simple crescent-shaped organelle that develops into a perinuclear ring with two small protrusions, followed by transformation into an extensive reticulated network as the parasite enlarges. Similarly, the apicoplast and the mitochondrion grow from single, small, discrete organelles into highly branched structures in later-stage parasites. These branched structures undergo an ordered fission - apicoplast followed by mitochondrion - to create multiple daughter organelles that are apparently linked as pairs for packaging into daughter cells. This is the first in-depth examination of intracellular organelles in live parasites during the asexual life cycle of this important human pathogen.

Animals↗

Phototropin is the blue-light receptor that controls multiple steps in the sexual life cycle of the green alga Chlamydomonas reinhardtii.

Blue light as an environmental cue plays a pivotal role in controlling the progression of the sexual life cycle in the green alga Chlamydomonas reinhardtii. Phototropin was considered a prime candidate for the blue-light receptor involved. By using the RNA interference method, knockdown strains with reduced phototropin levels were isolated. Those with severely reduced levels of this photoreceptor were partially impaired in three steps of the life cycle: in gametogenesis, the maintenance of mating ability, and the germination of zygotes. These observations suggest that phototropin is the principal sensory molecule used by this alga for the control of its life cycle by light.

Animals↗

The Schistosoma mansoni soluble proteome: a comparison across four life-cycle stages.

Differential analysis of immune responses to schistosomes has routinely been performed using complex mixtures of soluble proteins from various life-cycle stages, on the assumption that these differed significantly in composition. Proteomic techniques now allow us to characterise and compare such mixtures. The soluble proteins from cercariae, lung-schistosomula, adult worms and eggs of Schistosoma mansoni were separated by high-resolution two-dimensional electrophoresis and the resulting images analysed using appropriate software. A high degree of quantitative and qualitative similarity in spot pattern was revealed across the life-cycle, greatest between adjacent stages. To initiate mapping of these soluble proteomes, the 40 most abundant spots in each preparation, accounting for 21-46% of the total protein, were subjected to peptide fingerprinting by mass spectrometry. On average 55% of the spots were identified, but overall, these comprised only 32 different protein species. With one exception all proteins originated in the cytosol and 24 of the 32 had previously been pinpointed by virtue of their immunoreactivity, including four of the WHO priority vaccine candidates. The similarity in composition between the four preparations means that they are unlikely to discriminate adequately between immune responses to different life-cycle stages and argues strongly for the need to identify true stage-specific marker proteins. Equally, it is difficult to reconcile the abundance and immunogenicity of such cytosolic proteins with their status as vaccine candidates, as it is unlikely they will be accessible to the immune system in an intact parasite.

Animals↗

Further evidence on the internal life cycle of Przhevalskiana silenus (Diptera, Oestridae).

Knowledge of the internal life cycle of goat warble fly infestation is scarce despite ample data available on the aetiology, epidemiology, immunodiagnosis and treatment of such infestations. This study was carried out at the slaughterhouse of Rossano Calabro (Cosenza, southern Italy) on 154 animals from 10 months to 6 years of age from May 1997 to June 1998. 1206 Przhevalskiana silenus larvae were collected during the trial period from the subcutaneous tissue of the slaughtered animals. The larval stage average size ranged from 4.7 mm, for first instar larvae (May), to 16.6 mm, for third instar larvae (February), in the first cycle of infestation. No larvae were found in March-April, coinciding with the pupation period. Small first instar larvae were found at the beginning of the second cycle of infestation (May-June). Necroscopic examinations were also carried out on internal organs and no larvae were found. The results pointed out that the internal life cycle of P. silenus is exclusively subcutaneous and there is no internal migration of the larvae.

Age Factors↗

Life-cycle effects of single-walled carbon nanotubes (SWNTs) on an estuarine meiobenthic copepod.

Single-walled carbon nanotubes (SWNT) are finding increasing use in consumer electronics and structural composites. These nanomaterials and their manufacturing byproducts may eventually reach estuarine systems through wastewater discharge. The acute and chronic toxicity of SWNTs were evaluated using full life-cycle bioassays with the estuarine copepod Amphiascus tenuiremis (ASTM method E-2317-04). A synchronous cohort of naupliar larvae was assayed by culturing individual larvae to adulthood in individual 96-well microplate wells amended with SWNTs in seawater. Copepods were exposed to "as prepared" (AP) SWNTs, electrophoretically purified SWNTs, or a fluorescent fraction of nanocarbon synthetic byproducts. Copepods ingesting purified SWNTs showed no significant effects on mortality, development, and reproduction across exposures (p < 0.05). In contrast, exposure to the more complex AP-SWNT mixture significantly increased life-cycle mortality, reduced fertilization rates, and reduced molting success in the highest exposure (10 mg x L(-1)) (p < 0.05). Exposure to small fluorescent nanocarbon byproducts caused significantly increased life-cycle mortality at 10 mg x L(-1) (p < 0.05). The fluorescent nanocarbon fraction also caused significant reduction in life-cycle molting success for all exposures (p < 0.05). These results suggest size-dependent toxicity of SWNT-based nanomaterials, with the smallest synthetic byproduct fractions causing increased mortality and delayed copepod development over the concentration ranges tested.

Animals↗

Effects of four synthetic musks on the life cycle of the harpacticoid copepod Nitocra spinipes.

A full life-cycle (</=26 days exposure) toxicity test with the harpacticoid copepod Nitocra spinipes was used to study the effects of one nitro musk (musk ketone) as well as three polycyclic musks (Tonalide, Celestolide and Galaxolide). A subchronic individual life-table endpoint, the larval development rate, was recorded after 7-8 days exposure of juveniles and was significantly decreased in copepods exposed to sublethal concentrations of musk ketone, Celestolide and Galaxolide. However, none of the Tonalide concentrations had any effect on larval development. The lowest Galaxolide concentration (0.02 mg/l), which affected juvenile development, was about 100 times below the adult 96-h-LC(50)-value of 1.9 mg/l (95% confidence interval: 1.4-2.7 mg/l). However, none of the four musks had any agonistic or antagonistic activity in the ecdysteroid-sensitive Drosophila melanogaster B(II)-cell line. This indicates that the decrease in larval development rate was due to pharmacological effects rather than steroid receptor-mediated endocrine disruption. A modified Euler-Lotka equation was used to calculate a population-level endpoint, the intrinsic rate of natural increase (r(m)), from individual life-table endpoints, i.e. mortality rate, time of release of first brood, sex ratio, the fraction of ovigerous females among all females as well as the number of nauplii per ovigerous female. The second highest musk ketone concentration (0.1 mg/l) was the only treatment, which significantly affected r(m) (***P<0.001). At the highest musk ketone (0.3 mg/l) and Celestolide (0.3 mg/l) concentrations, all copepods were dead at the end of the exposures. This shows that a sensitive individual life-table endpoint is protective over the population-level endpoint r(m). Though we think that it is necessary to obtain population-level endpoints from standardised toxicity test, for ecologically successful risk characterisation of synthetic musks as well as other chemicals. The results from the present study show that it is possible to obtain population-level data from the full life-cycle test with N. spinipes. However, there seems to be little risk that synthetic musks are harmful to copepods at present environmental concentrations.

Animals↗

Assessing human health response in life cycle assessment using ED10s and DALYs: part 1--Cancer effects.

Life cycle assessment (LCA) is a framework for comparing products according to their total estimated environmental impact, summed over all chemical emissions and activities associated with a product at all stages in its life cycle (from raw material acquisition, manufacturing, use, to final disposal). For each chemical involved, the exposure associated with the mass released into the environment, integrated over time and space, is multiplied by a toxicological measure to estimate the likelihood of effects and their potential consequences. In this article, we explore the use of quantitative methods drawn from conventional single-chemical regulatory risk assessments to create a procedure for the estimation of the cancer effect measure in the impact phase of LCA. The approach is based on the maximum likelihood estimate of the effect dose inducing a 10% response over background, ED10, and default linear low-dose extrapolation using the slope betaED10 (0.1/ED10). The calculated effects may correspond to residual risks below current regulatory compliance requirements that occur over multiple generations and at multiple locations; but at the very least they represent a "using up" of some portion of the human population's ability to accommodate emissions. Preliminary comparisons are performed with existing measures, such as the U.S. Environmental Protection Agency's (U.S. EPA's) slope factor measure q1*. By analyzing bioassay data for 44 chemicals drawn from the EPA's Integrated Risk Information System (IRIS) database, we explore estimating ED10 from more readily available information such as the median tumor dose rate TD50 and the median single lethal dose LD50. Based on the TD50, we then estimate the ED10 for more than 600 chemicals. Differences in potential consequences, or severity, are addressed by combining betaED10 with the measure disability adjusted life years per affected person, DALYp. Most of the variation among chemicals for cancer effects is found to be due to differences in the slope factors (betaED10) ranging from 10(-4) up to 10(4) (risk of cancer/mg/kg-day).

Carcinogens, Environmental↗

Life cycle inventory analyses for five waste management options for discarded newspaper.

This paper presents the results of life cycle inventory (LCI) analyses that were carried out to determine the environmental impacts (emissions, resource extractions and land use) of different newspaper waste management options for the Helsinki Metropolitan Area (HMA). LCI analyses were performed for five product systems, in which discarded newspapers were divided into two streams: separately collected newspapers and newspapers in mixed waste. In all the options, the manufacturing and printing processes of newspaper were kept unchanged. The waste management alternatives included combinations of material recycling, energy recovery and landfilling. These product systems were modelled using the current collection rate of newspaper and four additional collection rates. The LCIs of the product systems showed that the life cycle phase causing the most environmental impacts was the paper mill. When comparing the different waste management systems, the energy recovery options were in general superior to landfilling. The ecological implications of the increased energy recovery and decreased material recycling of newspaper were, however, not yet considered in the study. These aspects were assessed in the life cycle impact assessment (LCIA), which was performed after the LCI phase.

Air Pollutants↗