Differing behavioral changes in crayfish and bluegill under short- and long-chain PFAS exposures: Field study in Northern Michigan, USA.

The emergent contaminant family, per- and poly-fluorinated alkyl substances (PFAS) has gained research attention due to their widespread detection and stability within the environment. Despite the growing amount of research on perfluorooctanesulfonic acid (PFOS) and perfluoro-n-octanoic acid (PFOA) in aquatic organisms, investigations detailing behavioral and physiological effects of aquatic organisms exposed to a mixture of PFAS analytes in the wild have been limited. The objective of this study was to evaluate the potential behavioral and histological effects of environmental exposure to PFAS compounds within multiple trophic levels of aquatic ecosystems. The current study investigates effects of environmentally relevant PFAS concentration exposures in crayfish (Faxonius immunis, F. rusticus, F. virilis) and bluegill (Lepomis macrochirus) sourced from four water bodies in Northern Michigan. Antipredator response and foraging behavioral assays were used to investigate potential effects on crayfish; a swimming speed behavioral assay and liver and gill histology analysis were used to investigate potential effects on fish. Linear mixed model and multiple regression analyses resulted in significant relationships between tissue accumulation levels of long chain PFAS compounds and crayfish foraging and fish critical swimming speed responses. Crayfish foraging decreased and fish critical swim speeds increased with PFAS exposure which may lead to energetic and population concerns. Antipredator response in crayfish and liver and gill histology in fish were not significantly related to PFAS tissue or water concentrations. The sensitivity of crayfish and bluegill behavior contributes to the growing body of research regarding the differential toxicity of short-chain and long-chain PFAS compounds. The sensitivity of some aquatic organism behaviors to PFAS accumulated in tissue may have implications for PFAS transfer and alterations to ecosystem functioning; based on the results of this field study, further laboratory research is recommended to further evaluate these relationships.

A Co-Association of Streptococcus mutans and Veillonella parvula/dispar in Root Caries Patients and In Vitro Biofilms.

Root caries in geriatric patients is a growing problem as more people are maintaining their natural teeth into advanced age. We determined the levels of various bacterial species previously implicated in root caries disease or health using quantitative real-time PCR in a pilot study of 7 patients with 1 to 4 root caries lesions per person. Levels of 12 different species on diseased roots compared to healthy (contralateral control) roots were measured. Four species were found at significantly higher levels on diseased roots (Streptococcus mutans, Veillonella parvula/dispar, Actinomyces naeslundii/viscosus, and Capnocytophaga granulosa) compared across all plaque samples. The level of colonization by these species varied dramatically (up to 1,000-fold) between patients, indicating different patients have different bacteria contributing to root caries disease. Neither of the two species previously reported to correlate with healthy roots (C. granulosa and Delftia acidovorans) showed statistically significant protective roles in our population, although D. acidovorans showed a trend toward higher levels on healthy teeth (P = 0.08). There was a significant positive correlation between higher levels of S. mutans and V. parvula/dispar on the same diseased teeth. In vitro mixed biofilm studies demonstrated that co-culturing S. mutans and V. parvula leads to a 50 to 150% increase in sucrose-dependent biofilm mass compared to S. mutans alone, depending on the growth conditions, while V. parvula alone did not form in vitro biofilms. The presence of V. parvula also decreased the acidification of S. mutans biofilms when grown in artificial saliva and enhanced the health of mixed biofilms.

Environmentally relevant atrazine exposure leads to increases in DNA damage and changes in morphology in the hepatopancreas of crayfish (Faxonius virilis).

The herbicide atrazine is widely used for controlling broad leaf weeds and increasing crop yields in agricultural areas. Atrazine enters aquatic environments through runoff, ground water discharge and seepage where concentrations have been recorded above 300 ppb. Exposure to the herbicide atrazine at environmentally relevant concentrations has been shown to negatively impact aquatic organisms, including crayfish. Because xenobiotics are concentrated in the crayfish hepatopancreas (digestive gland), we examined changes in morphology and DNA damage in hepatopancreatic tissue structure and cells following a 10-day exposure to atrazine (0, 10, 40, 80, 100 and 300 ppb). We found that there were marked morphological changes, post-exposure, for all atrazine concentrations tested. Hepatopancreatic tissue exhibited degenerated tubule epithelium with necrosis of microvilli, tubule lumen dilation, changes in tubular epithelium height and vacuolization of the epithelium. Likewise, we also performed a terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labeling (TUNEL) assay which showed the percentage of cells with DNA damage increased following atrazine exposure. Crayfish hepatopancreatic tissue displayed significant increases in TUNEL-positive cells following exposure to atrazine at 100 ppb and above. Overall, exposure to atrazine at environmentally relevant concentrations damages hepatopancreatic tissue. This impairment could lead to changes in biotransformation, detoxification, digestion and molting, subsequently reducing crayfish populations and negatively impacting the aquatic ecosystem.

Environmentally relevant atrazine exposures cause DNA damage in cells of the lateral antennules of crayfish (Faxonius virilis).

The herbicide atrazine is heavily applied in agricultural areas in the Midwestern United States and can run-off and seep into surrounding aquatic habitats where concentrations can reach over 300 ppb. It is known that acute exposures to 80 ppb atrazine cause lasting deficiencies in the chemoreception of food and mate odors. Since atrazine impairs chemosensory responses, the goal of this study was to determine the effect of atrazine on cells, including olfactory sensory neurons, located in the lateral antennules of crayfish. In this experiment, we treated crayfish for 10 days with ecologically relevant concentrations of 0, 10, 40, 80, 100 and 300 ppb (µg L-1) of atrazine. Following treatments, the distal portion of the lateral antennules was cryosectioned. We used a TdT mediated dUTP nick-end labeling (TUNEL) assay to determine if any cells had DNA damage and may be thus undergoing apoptosis. We found that as atrazine concentrations increase above 10 ppb, the number of TUNEL-positive cells, visualized in the lateral antennules, significantly increases. Our data show that atrazine exposure causes DNA damage in cells of the lateral antennules, including olfactory sensory neurons, thus leading to impairments in chemosensory abilities. Because crayfish rely heavily on chemoreception for survival, changes in their ability to perceive odors following atrazine exposure may have detrimental effects on population size.

Cytochrome P450 and Glutathione-S-Transferase Activity are Altered Following Environmentally Relevant Atrazine Exposures in Crayfish (Faxoniusvirilis).

The herbicide atrazine is heavily applied in the U.S. Midwest to control broadleaf weeds. It enters local streams and rivers through runoff and seepage, and exposure can affect non-target aquatic organisms, like crayfish. We examined sublethal effects of atrazine on the expression and activity of the detoxification enzymes cytochrome P450 (CYP450) and glutathione-S-transferase (GST) in crayfish. Crayfish were exposed to 0, 10, 40, 80, 100 and 300 ppb atrazine for 1, 2, 4, 7 and 10 days. Their hepatopancreas was collected and CYP450 expression and GST activity was analyzed. Atrazine exposure caused differential expression and activity of CYP450 and GST. CYP450 expression varied over exposure concentrations and time. Further, GST activity significantly increased following a 2 day, 10 ppb exposure to atrazine and a 300 ppb atrazine exposure for all days tested. We found that atrazine detoxification is a dynamic process that changes with the length and intensity of atrazine exposure.