Swee J. Teh

Ingested plastic transfers hazardous chemicals to fish and induces hepatic stress

Plastic debris litters aquatic habitats globally, the majority of which is microscopic (< 1 mm), and is ingested by a large range of species. Risks associated with such small fragments come from the material itself and from chemical pollutants that sorb to it from surrounding water. Hazards associated with the complex mixture of plastic and accumulated pollutants are largely unknown. Here, we show that fish, exposed to a mixture of polyethylene with chemical pollutants sorbed from the marine environment, bioaccumulate these chemical pollutants and suffer liver toxicity and pathology. Fish fed virgin polyethylene fragments also show signs of stress, although less severe than fish fed marine polyethylene fragments. We provide baseline information regarding the bioaccumulation of chemicals and associated health effects from plastic ingestion in fish and demonstrate that future assessments should consider the complex mixture of the plastic material and their associated chemical pollutants.

Policy: Classify plastic waste as hazardous

Policies for managing plastic debris are outdated and threaten the health of people and wildlife, say Chelsea M. Rochman, Mark Anthony Browne and colleagues.

Histopathologic biomarkers in feral freshwater fish populations exposed to different types of contaminant stress

Histopathologic alterations of gill, liver, and spleen were studied in feral fish from three freshwater ecosystems that experience different types of contaminant stress. East Fork Poplar Creek (EFPC), a third-order stream in East Tennessee, receives point source discharges of mixed contaminants from a nuclear weapons facility located near its headwater. The Pigeon River (PR), a high-gradient fifth order stream, is impacted by bleached kraft mill effluent (BKME). Hartwell Reservoir (HR), a US Army Corp of Engineers impoundment of the Savannah River, contains high levels of PCBs in sediment and biota. Brushy Fork Creek (BFC), the Little River (LR), and the Tugaloo River (TR) are relatively free of contaminants, and served as reference sites for the three respective ecosystems of this study. Certain organ and tissue lesions, detected microscopically, were common to fish from both reference and contaminated sites. These included parasites, inflammation, glycogen deficiency, macrophage aggregates (MA), and diffuse fatty change in the liver; parasites and MA in the spleen; and parasites, secondary lamellar fusion, and variable epithelial cell hyperplasia in the gills. Lesions found only in fish from contaminated sites were: (1) cholangiomas in liver of redbreast sunfish (Lepomis auratus) collected from EFPC; (2) amphophilic and eosinophilic foci of cellular alteration, diffuse biliary preductular and ductular hyperplasia with islands of hyperplastic basophilic hepatocytes, and two metastatic thyroidal carcinomas in spleen of redbreast sunfish from PR; (3) severe lipidosis, vacuolated and basophilic foci in largemouth bass (Micropterus salmoides) from HR; (4) splenic lymphoid cell depletion and vascular congestion, with necrosis of reticuloendothelial cells in fish collected from EFPC and HR; (5) hyperplasia of mucous and chloride cells, deformed branchial cartilage, severe and diffuse aneurysms of lamellae, and edema at the base of the secondary lamellae were in gill of fish from all three sites. The finding of specific lesions only in fish from contaminated sites suggests a contaminant etiology. Histopathology biomarker lesions identified in this study are similar to those observed in laboratory exposures of fishes to specific pollutants. Further refinement of these biomarker approaches will be discussed in light of multiple stressors and their effects.

Selenium biotransformations into proteinaceous forms by foodweb organisms of selenium-laden drainage waters in California

Selenium contamination represents one of the few clear cases where environmental pollution has led to devastation of wildlife populations, most notably in agricultural drainage evaporation and power plant coal-fly ash receiving ponds. Complex biogeochemistry, in particular extensive biotransformations and foodchain transfer, governs Se ecotoxicology and toxicology, for which the mechanism(s) are still elusive. However, total waterborne Se concentration has been widely used as a criterion for regulating and mitigating Se risk in aquatic ecosystems, which does not account for Se biogeochemistry and its site-dependence. There is a need for more reliable indicator(s) that encompass Se ecotoxicity and/or toxicity. Selenomethionine warrants special attention since it simulates Se toxicosis of wildlife in laboratory feeding studies. While low in free selenomethionine, microphytes isolated from Se-laden agricultural evaporation ponds were abundant in proteinaceous selenomethionine. This prompted a more extensive survey of Se speciation in foodchain organisms including microphytes, macroinvertebrates, fish, and bird embryos residing mainly in the agricultural drainage systems of the San Joaquin Valley, California. Total Se in biomass, water-soluble fractions, and protein-rich fractions were measured along with GC-MS analysis of proteinaceous selenomethionine. In all foodchain organisms, water-soluble Se constituted the major fraction of total biomass Se, while proteinaceous Se was a substantial, if not dominant, fraction of the water-soluble Se. In turn, proteinaceous selenomethionine comprised an important fraction of proteinaceous Se. In terms of total biomass Se, an average 1400-fold of Se biomagnification from water to microphytes was observed while subsequent transfer from microphytes to macroinvertebrates exhibited an average of only 1.9-fold. The latter transfer was more consistent and greater in extent for proteinaceous Se and proteinaceous selenomethionine, which is consistent with their importance in foodchain transfer. Proteinaceous Se in the omnivorous carp (Cyprinus carpio) liver also demonstrated a relation to ovarian lesions, while deformed stilt (Himantopus mexicanus) embryo was more abundant in proteinaceous selenomethionine than were normal embryos. Although limited in the number of organisms surveyed, these findings provide an impetus for further field and laboratory feeding studies to substantiate the hypothesis that proteinaceous selenomethionine underlies Se ecotoxicity, which may in turn prove to be a reliable indicator of Se risk in aquatic ecosystems.

Anthropogenic debris in seafood: Plastic debris and fibers from textiles in fish and bivalves sold for human consumption

The ubiquity of anthropogenic debris in hundreds of species of wildlife and the toxicity of chemicals associated with it has begun to raise concerns regarding the presence of anthropogenic debris in seafood. We assessed the presence of anthropogenic debris in fishes and shellfish on sale for human consumption. We sampled from markets in Makassar, Indonesia, and from California, USA. All fish and shellfish were identified to species where possible. Anthropogenic debris was extracted from the digestive tracts of fish and whole shellfish using a 10% KOH solution and quantified under a dissecting microscope. In Indonesia, anthropogenic debris was found in 28% of individual fish and in 55% of all species. Similarly, in the USA, anthropogenic debris was found in 25% of individual fish and in 67% of all species. Anthropogenic debris was also found in 33% of individual shellfish sampled. All of the anthropogenic debris recovered from fish in Indonesia was plastic, whereas anthropogenic debris recovered from fish in the USA was primarily fibers. Variations in debris types likely reflect different sources and waste management strategies between countries. We report some of the first findings of plastic debris in fishes directly sold for human consumption raising concerns regarding human health.

Early warning signs of endocrine disruption in adult fish from the ingestion of polyethylene with and without sorbed chemical pollutants from the marine environment

Plastic debris is associated with several chemical pollutants known to disrupt the functioning of the endocrine system. To determine if the exposure to plastic debris and associated chemicals promotes endocrine-disrupting effects in fish, we conducted a chronic two-month dietary exposure using Japanese medaka (Oryzias latipes) and environmentally relevant concentrations of microplastic (<1mm) and associated chemicals. We exposed fish to three treatments: a no-plastic (i.e. negative control), virgin-plastic (i.e. virgin polyethylene pre-production pellets) and marine-plastic treatment (i.e. polyethylene pellets deployed in San Diego Bay, CA for 3 months). Altered gene expression was observed in male fish exposed to the marine-plastic treatment, whereas altered gene expression was observed in female fish exposed to both the marine- and virgin-plastic treatment. Significant down-regulation of choriogenin (Chg H) gene expression was observed in males and significant down-regulation of vitellogenin (Vtg I), Chg H and the estrogen receptor (ERα) gene expression was observed in females. In addition, histological observation revealed abnormal proliferation of germ cells in one male fish from the marine-plastic treatment. Overall, our study suggests that the ingestion of plastic debris at environmentally relevant concentrations may alter endocrine system function in adult fish and warrants further research.

Determining the sensitive developmental stages of intersex induction in medaka (Oryzias latipes) exposed to 17β-estradiol or testosterone

Certain environmentally persistent compounds can adversely affect reproduction by acting as steroid hormone agonists or antagonists. The goal of the present study was to determine the developmental stage most susceptible to exogenous hormone (estradiol and testosterone) exposure using a small teleost model. In the first (pilot study) of two experiments, medaka (Oryzias latipes), at varying developmental stages, were bath-exposed to 5 micrograms/l 17 beta-estradiol for 24 h. At 5 months of age, fecundity, fertility and embryo and larval viability (reproductive success) were investigated in control and exposed groups. Fish at 1, 1.5, 2 and 5.5 months of age were also sampled, processed and examined histologically for gonadal alteration. No significant differences in mortality, gonadal morphology, body weight, sex-ratio or time to maturity were seen between control and exposed fish. At 5 months, however, when exposure groups were compared to controls, significant differences were seen in reproductive success and viability of offspring. A second experiment exposed embryo stage 10, and 1-, 7- and 21-day-old larvae for 6 days to 15 micrograms/l 17 beta-estradiol or 100 micrograms/l testosterone. No significant differences were seen at 5 months in mortality, body weight, or time to sexual maturity. However, sex-ratios were significantly biased toward female in the stage 10, 1- and 7-day post-hatch estradiol exposure groups. No significant changes in sex-ratio were associated with testosterone exposure at any developmental stage. Further, intersex gonads were observed in fish from all groups exposed to 15 micrograms/l estradiol. Only those fish exposed as newly hatched fry or at 1 week post-hatch displayed intersex gonads following 100 micrograms/l testosterone exposure. Data from these experiments show that newly hatched fry are that life stage most sensitive to hormone exposure and the most appropriate to use in determining effects of known endocrine-disrupting compounds.

Histological and histochemical changes in the digestive tract of white sturgeon larvae during ontogeny

Ontogenetic changes in digestive tract histology and digestive enzyme histochemistry were investigated 11 to 36 days post-hatch in white sturgeon Acipenser transmontanus larvae. From initiation of exogenous feeding (12 days post-hatch), larvae were fed a commercial salmonid diet for the ensuing 24 days. The digestive system of white sturgeon displayed a high degree of morphologic organization and functionality at the onset of exogenous feeding. An enhancement of digestive capacities occurred with transition to active feeding. On day 2 of feeding, there was a clear increase of alkaline phosphatase, aminopeptidase M, dipeptidyl peptidase IV, and γ-glutamyl transpeptidase activity in the brush border of the spiral intestine. This strong activity is an apparent confirmation of the importance of this segment of the intestine for protein digestion and nutrient absorption. The functional development of the pyloric intestine occurred on day 4 and was concomitant with an increase in the activity of brush border and cytoplasmic enzymes such as acetylcholinesterase, dipeptidyl peptidase II, α- and β-galactosidases. The absence of acetylcholinesterase, lactase, nonspecific esterase, and weak activity of exopeptidases and alkaline phosphatase in the anterior intestine suggests that this segment of the intestine may be less important in nutrient absorption than the pyloric and spiral intestines. The observed quantitative and qualitative differences in enzyme activity along the intestine indicate a high degree of specialization of each segment for specific digestive and absorptive processes.

Isolated trout liver cells: Establishing short-term primary cultures exhibiting cell-to-cell interactions

SummaryComposition and interactions of cell types in rainbow trout (Oncorhynchus mykiss) liver digested with collagenase and cultured in serum-free media were investigated. Suspensions obtained after digesting trout liver with collagenase contained all the cell types present in the liver, including liver parenchymal cells (hepatocytes), biliary epithelial cells, sinusoidal endothelium, fat-storing cells of Ito, and macrophages. A major cell pellet, mainly hepatocytes but containing significant numbers of biliary epithelial cells, was obtained by centrifuging the cell suspension at 120×g for 1 min. Cells present in this pellet quantitatively attached to culture plates coated with a trout skin extract and remain attached for 4 to 6 d with good retention of intracellular enzymes and DNA. When in culture, significant changes in and among the cells were observed. Initial preparations were rounded, single cells. Within several hours, however, cellular interactions leading to aggregation became evident and aggregates increased in size for 2 to 3 d. Scanning electron microscopy (EM) showed frequent shaftlike projections from margins of the aggregates. Transmission EM indicated that these projections represent biliary ductules forming in vitro. Adjacent hepatocytes also showed plasma membrane specializations forming junctional complexes and canaliculi characteristics of normal trout liver. After 5 to 6 d in culture, significant numbers of the cell aggregates dislodged from the plate. Analysis showed the dislodged cells were viable but vacuolated. The reestablishment in vitro of morphologic relationships resembling in situ tissue components suggest these culture preparations may have significant utility in cooperative metabolic studies of cell interactions in trout liver.

Long-term sorption of metals is similar among plastic types: implications for plastic debris in aquatic environments.

Concerns regarding plastic debris and its ability to accumulate large concentrations of priority pollutants in the aquatic environment led us to quantify relationships between different types of mass-produced plastic and metals in seawater. At three locations in San Diego Bay, we measured the accumulation of nine targeted metals (aluminum, chromium, manganese, iron, cobalt, nickel, zinc, cadmium and lead) sampling at 1, 3, 6, 9 and 12 months, to five plastic types: polyethylene terephthalate (PET), high-density polyethylene (HDPE), polyvinyl chloride (PVC), low-density polyethylene (LDPE), and polypropylene (PP). Accumulation patterns were not consistent over space and time, and in general all types of plastic tended to accumulate similar concentrations of metals. When we did observe significant differences among concentrations of metals at a single sampling period or location in San Diego Bay, we found that HDPE typically accumulated lesser concentrations of metals than the other four polymers. Furthermore, over the 12-month study period, concentrations of all metals increased over time, and chromium, manganese, cobalt, nickel, zinc and lead did not reach saturation on at least one plastic type during the entire 12-month exposure. This suggests that plastic debris may accumulate greater concentrations of metals the longer it remains at sea. Overall, our work shows that a complex mixture of metals, including those listed as priority pollutants by the US EPA (Cd, Ni, Zn and Pb), can be found on plastic debris composed of various plastic types.