Bioaccumulation and metabolic impact of environmental PFAS residue on wild-caught urban wetland tiger snakes (Notechis scutatus).

PFAS contamination of urban waters is widespread but understanding the biological impact of its accumulation is limited to humans and common ecotoxicological model organisms. Here, we combine PFAS exposure and bioaccumulation patterns with whole organism responses and omics-based ecosurveillance methods to investigate the potential impacts of PFAS on a top predator of wetlands, the tiger snake (Notechis scutatus). Tiger snakes (18 male and 17 female) were collected from four wetlands with varying PFAS chemical profiles and concentrations in Perth, Western Australia. Tiger snake livers were tested for 28 known PFAS compounds, and Σ28PFAS in liver tissues ranged between 322 ± 193 µg/kg at the most contaminated site to 1.31 ± 0.86 µg/kg at the least contaminated site. The dominant PFAS compound detected in liver tissues was PFOS. Lower body condition was associated with higher liver PFAS, and male snakes showed signs of high bioaccumulation whereas females showed signs of maternal offloading. Biochemical profiles of snake muscle, fat (adipose tissue), and gonads were analysed using a combination of liquid chromatography triple quadrupole (QqQ) and quadrupole time-of-flight (QToF) mass spectrometry methodologies. Elevated PFAS was associated with enriched energy production and maintenance pathways in the muscle, and had weak associations with energy-related lipids in the fat tissue, and lipids associated with cellular genesis and spermatogenesis in the gonads. These findings demonstrate the bioavailability of urban wetland PFAS in higher-order reptilian predators and suggest a negative impact on snake health and metabolic processes. This research expands on omics-based ecosurveillance tools for informing mechanistic toxicology and contributes to our understanding of the impact of PFAS residue on wildlife health to improve risk management and regulation.

Snake scales record environmental metal(loid) contamination.

Wetland snakes, as top predators, are becoming globally recognised as bioindicators of wetland contamination. Livers are the traditional test organ for contaminant exposure in organisms, but research is moving towards a preference for non-lethal tissue sampling. Snake scales can be used as an indicator of exposure, as many metals bind to the keratin. We used laser ablation with inductively coupled plasma-atomic emission spectroscopy and mass spectrometry (LA-ICP-MS) to quantify the concentrations of 19 metals and metalloids (collectively referred to 'metals' hereafter) in Western tiger snake (Notechis scutatus occidentalis) scales from four wetlands along an urban gradient, and compared them to concentrations measured in captive tiger snake scales. We conducted repeat measures to determine the concentration accuracy of each metal using LA-ICP-MS. Concentrations in wild Western tiger snake scales were significantly higher than in reference tiger snake scales for most metals analysed, suggesting accumulation from environmental exposure. We compared the scale concentrations to sediment concentrations of sampled wetlands, and found inter-site differences between mean concentrations of metals in scales parallel patterns recorded from sediment. Four metals (Mn, As, Se, Sb) had strong positive correlations with liver tissue contents suggesting scale concentrations can be used to infer internal concentrations. By screening for a larger suite of metals than we could using traditional digestive methods, we identified additional metals (Ti, V, Sr, Cs, Tl, Th, U) that may be accumulating to levels of concern in tiger snakes in Perth, Western Australia. This research has progressed the use of LA-ICP-MS for quantifying a suite of metals available in snake scales, and highlights the significance of using wetland snake scales as a non-lethal indicator of environmental contamination.

The Broad-Scale Analysis of Metals, Trace Elements, Organochlorine Pesticides and Polycyclic Aromatic Hydrocarbons in Wetlands Along an Urban Gradient, and the Use of a High Trophic Snake as a Bioindicator.

Wetlands and their biodiversity are constantly threatened by contaminant pollution from urbanisation. Despite evidence suggesting that snakes are good bioindicators of environmental health, the bioaccumulation of contaminants in reptiles is poorly researched in Australia. We conducted the first broad-scale analysis of 17 metals and trace elements, 21 organochlorine pesticides, and 14 polycyclic aromatic hydrocarbons in the sediments (4 samples per site, December 2018) from four wetlands along an urban gradient in Perth, Western Australia, and from the livers (5 livers per site, February-April 2019) of western tiger snakes Notechis scutatus occidentalis captured at those sites. All 17 metals and trace elements were detected in the sediments of wetlands as well as 16 in the livers of tiger snakes. Arsenic, Cu, Hg, Pb, Se, and Zn were at concentrations exceeding government trigger values in at least one sediment sample. Two organochlorine pesticides and six of seven polycyclic aromatic hydrocarbons were detected in the sediments of a single wetland, all exceeding government trigger values, but were not detected in tiger snakes. Metals and trace elements were generally in higher concentration in sediments and snake livers from more heavily urbanised wetlands. The least urbanised site had some higher concentrations of metals and trace elements, possibly due to agriculture contaminated groundwater. Concentrations of nine metals and trace elements in snake livers were statistically different between sites. Arsenic, Cd, Co, Hg, Mo, Sb, and Se near paralleled the pattern of contamination measured in the wetland sediments; this supports the use of high trophic wetland snakes, such as tiger snakes, as bioindicators of wetland contamination. Contamination sources and impacts on these wetland ecosystems and tiger snakes are discussed herein.

Integrating Multiple Biomarkers of Fish Health: A Case Study of Fish Health in Ports.

Biomarkers of fish health are recognised as valuable biomonitoring tools that inform on the impact of pollution on biota. The integration of a suite of biomarkers in a statistical analysis that better illustrates the effects of exposure to xenobiotics on living organisms is most informative; however, most published ecotoxicological studies base the interpretation of results on individual biomarkers rather than on the information they carry as a set. To compare the interpretation of results from individual biomarkers with an interpretation based on multivariate analysis, a case study was selected where fish health was examined in two species of fish captured in two ports located in Western Australia. The suite of variables selected included chemical analysis of white muscle, body condition index, liver somatic index (LSI), hepatic ethoxyresorufin-O-deethylase activity, serum sorbitol dehydrogenase activity, biliary polycyclic aromatic hydrocarbon metabolites, oxidative DNA damage as measured by serum 8-oxo-dG, and stress protein HSP70 measured on gill tissue. Statistical analysis of individual biomarkers suggested little consistent evidence of the effects of contaminants on fish health. However, when biomarkers were integrated as a set by principal component analysis, there was evidence that the health status of fish in Fremantle port was compromised mainly due to increased LSI and greater oxidative DNA damage in fish captured within the port area relative to fish captured at a remote site. The conclusions achieved using the integrated set of biomarkers show the importance of viewing biomarkers of fish health as a set of variables rather than as isolated biomarkers of fish health.

Field studies using fish biomarker--how many fish are enough?

Significant efforts are invested in field studies using fish, and it is important to optimize the number of organisms collected to evaluate the possible impacts of contamination. This paper provides ecotoxicologists with the approximate numbers of fish needed to identify statistically significant differences among samples using physiological indices and biochemical markers of fish health. The numbers of fish to collect are reported for ethoxyresorufin-o-deethylase (EROD); ethoxycoumarin-o-deethylase (ECOD), serum sorbitol dehydrogenase (sSDH), stress proteins, gonadosomatic index, liver somatic index, condition factor, and biliary metabolites of polycyclic aromatic hydrocarbons. The number of fish to collect was as few as four for ECOD activity (with a power of 80%), but as high as 106 for CF (with a power of 95%). Achieving statistical significance between sites does not help in the interpretation of the biological significance of a parameter, but well-planned field samplings will maximize the chances of correctly identifying areas of concern.

Factors associated with hallux valgus in a population-based study of older women and men: the MOBILIZE Boston Study.

OBJECTIVE: To examine potential risk factors for hallux valgus in community-dwelling elders. METHOD: Data from 600 MOBILIZE Boston Study participants (386 women and 214 men) were analyzed. Hallux valgus was defined as >15 degrees angular deviation of the hallux with respect to the first metatarsal bone toward the lesser toes. Associations of hallux valgus with age, body mass index (BMI), race, education, pes planus, foot pain, and in women, history of high heel shoe use, were assessed using sex-specific Poisson regression with robust variance estimation for risk ratios (RR) and 95% confidence intervals (CI). RESULTS: Hallux valgus was present in 58% of women and 25% of men. Higher BMI was inversely associated with presence of hallux valgus in women (P trend=0.001), with the strongest inverse association observed in those with BMI of 30.0 or more compared to those with normal BMI (RR=0.7, 95% CI: 0.5, 0.9). Women, who usually wore high-heeled shoes during ages 20-64 years compared to those who did not, had increased likelihood of hallux valgus (RR=1.2, 95% CI: 1.0, 1.5). Among men, those with BMI between 25.0 and 29.9 had increased likelihood of hallux valgus compared to those with normal BMI (RR=1.9, 95% CI: 1.0, 3.5). Men with pes planus were more likely to have hallux valgus (RR=2.1, 95% CI: 1.3, 3.3) compared to men without pes planus. CONCLUSION: In women, hallux valgus was associated with lower BMI and high heel use during ages 20-64, while in men, associations were observed with higher BMI and pes planus. Our results suggest that the etiologic mechanisms for hallux valgus may differ between men and women.

Alterations of metabolic enzymes in Australian bass, Macquaria novemaculeata, after exposure to petroleum hydrocarbons.

Australian bass Macquaria novemaculeata were exposed to the water-accommodated fraction of Bass Strait crude oil, dispersed crude oil, or burnt crude oil to assess sublethal effects of oil spill remediation techniques on fish. Fish were exposed to these treatments for 16 days either through the water column or by way of a pre-exposed diet of amphipod Allorchestes compressa. Fish gills, liver, and white muscle were sampled and cytochrome C oxidase (CCO) and lactate dehydrogenase (LDH) activities quantified. In all treatments containing fish exposed by way of the water column, aerobic activity increased in the gills, whereas a decrease of this enzymic activity was observed in the liver and white muscle. Exposures by way of the food pathway indicated similar trends. Anaerobic (LDH) activity increased in the gills, liver, and white muscle after waterborne exposures. Stimulation in anaerobic activity also occurred in the liver and white muscle of fish after exposure to contaminated food. CCO activity in the gills was the most sensitive biomarker when monitoring waterborne exposures to petroleum hydrocarbons. In the gills, the dispersed oil treatment resulted in the most pronounced biological response, suggesting that in the short term the use of dispersants on an oil slick might cause the most perturbations to fish metabolism.

Pink snapper (Pagrus auratus) as a bioindicator of aquatic. Environmental health in Western Australia.

Pink snapper (Pagrus auratus), an endemic Western Australian fish species, was tested for its potential as a bioindicator of aquatic environmental health. Mixed function oxygenase (MFO) induction (as a biomarker of exposure), and sorbitol dehydrogenase (SDH) activity (as a biomarker of liver damage) were of special interest to the study as these biochemical tools have not been validated for any Western Australian fish species. Juvenile pink snapper were injected intraperitoneally (i.p.) with 0, 10, 100, 500, and 1000 micrograms PCB-126 per kilogram. Fish were sacrificed 10 days postinjection, and livers and blood samples were collected for MFO and SDH analysis, respectively. Doses of 10 and 100 micrograms PCB-126 per kilogram caused the highest MFO induction, while doses of 0 and 1000 micrograms PCB-126 per kilogram did not result in higher MFO activity relative to carrier-injected control fish. SDH activities were not significantly different among treatments indicating that hepatocellular damage was not responsible for the reduced MFO activity at the highest dose. The result of the study demonstrates that pink snapper may potentially be used as a bioindicator species in Western Australia when MFO is used as a biomarker.

The effect of different oil spill remediation techniques on petroleum hydrocarbon elimination in Australian bass (Macquaria novemaculeata).

Petroleum hydrocarbons were investigated in juvenile Australian bass, Macquaria novemaculeata, following exposure to the water accommodated fraction (WAF) of Bass Strait crude oil, chemically dispersed crude oil, and burnt crude oil. Each treatment was administered for 16 days either through the water column or through the diet (amphipod, Allorchestes compressa). Polycyclic aromatic hydrocarbon (PAH) elimination was determined by measuring biliary benzo(a)pyrene (B(a)P) and naphthalene-type metabolites. Biliary PAH-type metabolite concentrations varied with the type of oil spill remediation technique, route of exposure (food versus water), and exposure concentration. Fish exposed to chemically dispersed crude oil via the water exhibited the highest PAH-type biliary metabolite concentrations, relative to fish exposed to other treatments. In fish exposed via the diet, the highest concentration of both types of biliary metabolites also appeared in the dispersed oil-exposed individuals. The results suggest that chemically dispersing oil may have the greatest effect on bioavailability of hydrocarbons, both through waterborne and food chain exposures.

Metabolic responses of fish following exposure to two different oil spill remediation techniques.

To assess the impacts of two oil spill remediation techniques on fish metabolism, change in aerobic and anaerobic enzyme activities in juvenile Australian Bass, Macquaria novemaculeata, was examined. Changes in cytochrome C oxidase (CCO) and lactate dehydrogenase (LDH) activities were investigated following exposure to the crude oil water accommodated fraction (WAF) and chemically dispersed crude oil WAF. There was a significant stimulation in CCO activity in the gills and livers of fish exposed to the WAF of Bass Strait crude oil and chemically dispersed crude oil, compared to the control treatment. In addition, LDH activity was significantly stimulated in the liver of fish exposed to dispersed crude oil WAF, compared to the crude oil WAF. Fish exposed to the dispersed crude oil WAF treatment had significantly higher oxygen consumption, as measured by oxygen depletion in a sealed chamber, than fish exposed to the crude oil WAF and control treatments.

Autonomic and neurohumoral control of postprandial blood pressure in healthy aging.

BACKGROUND: Postprandial hypotension (PPH) is a common and morbid problem in elderly people that is associated with an impaired vascular response to meal digestion. Healthy aging in the absence of blood pressure elevation is associated with autonomic and neurohumoral changes that may influence the vascular response to meal ingestion. However, it is not known whether these age-related changes are associated with the development of PPH. METHODS: We measured hemodynamic (blood pressure, heart rate, and forearm vascular resistance), autonomic (power spectral analysis of heart rate and blood pressure variability), and neurohumoral (plasma norepinephrine, renin, aldosterone, and endothelin) responses to a mixed 425 kilocalorie (kcal) meal in 89 rigorously screened healthy subjects aged 20-39, 40-59, and 60-83 years. RESULTS: After the meal, supine mean arterial blood pressure fell significantly only in the middle-aged group by 5.4 +/- 7.9 mm Hg at 30 minutes (p = .02). Forearm vascular resistance fell after the meal in all age groups ( p = .0001). Older groups had higher plasma norepinephrine (p = .02), lower heart rate (p = .03), lower cardiovagal activity (p = .0001), and lower sympathetic vasomotor (p = .000) activity, but there was no difference in the response of these variables to a meal. CONCLUSION: Healthy aging, in the absence of blood pressure elevation, alters the level of autonomic activity without further impairing the ability to maintain blood pressure during meal digestion. Hemodynamic, autonomic, and neurohumoral responses to meal ingestion remain unchanged in very healthy, normotensive elderly adults.

EROD induction and biliary metabolite excretion following exposure to the water accommodated fraction of crude oil and to chemically dispersed crude oil.

Immature Atlantic salmon (Salmo salar) were exposed to water accommodated fraction (WAF) of Bass Strait crude oil or to Corexit 9527-dispersed crude oil for 6 days, followed by a depuration period of 29 days. Serum sorbitol dehydrogenase (SDH) levels, indicator of liver damages, remained low during the experiment. Hepatic EROD activity was induced within 2 days following the onset of the exposure in both treatments, and persisted for 2-4 and 4-6 days after transfer to clean sea water in the WAF and dispersed oil treatment, respectively. Naphthalene-type metabolites, determined by fixed-wavelength fluorescence detection, appeared in the bile of the fish with 2 days' delay compared to EROD induction. In both treatments, EROD activity induction and levels of naphthalene-type metabolites in the bile were significantly related. The biliary levels of naphthalene-type metabolites were over 15 times higher in fish exposed to dispersed crude oil relative to fish exposed to the WAF of Bass Strait crude oil. BaP-type metabolites appeared only in the bile of the fish exposed to the WAF, possibly due to BaP-type compounds remaining associated with the dispersant in the water column or to an inhibition of Phase II detoxification enzymes by the dispersant. Bile metabolites as determined by fixed-wavelength fluorescence and EROD induction appear to be sensitive and complementary biomarkers of exposure to PAH.

Effects of postprandial walking exercise on meal-related hypotension in frail elderly patients.

The purpose of this study was to assess the hemodynamic effects of a postmeal walk in frail elderly patients with postprandial hypotension. We demonstrated that frail elderly patients with postprandial hypotension are able to increase their blood pressure and heart rate in response to a postmeal walking exercise, but this effect is limited to the exercise period only and is not sustained during subsequent rest.

Metabolic enzyme activities in fish gills as biomarkers of exposure to petroleum hydrocarbons.

Metabolic effects of low-level exposure of Atlantic salmon (Salmo salar) to the water accommodated fraction (WAF) of crude oil and to dispersed crude oil were studied. Aerobic enzymes citrate synthase and cytochrome C oxidase, and anaerobic enzyme lactate dehydrogenase were measured in gills during a 4-day exposure to low concentrations of dispersed Bass Strait crude oil and WAF, and during the following 8 days of depuration in clean seawater. Relative to pre-exposure levels, citrate synthase and lactate dehydrogenase exhibited a significant inhibition of activity during exposure to the WAF of crude oil, and to dispersed crude oil, while activity of cytochrome C oxidase remained unchanged. Citrate synthase activities returned to preexposure levels after 4 days following termination of exposure for the WAF-exposed fish, and after 2 days for the dispersed-oil-exposed fish. After the termination of exposure to both treatments, lactate dehydrogenase activity remained low relative to levels measured prior to exposure, which indicated that the activity of this enzyme may be a sensitive medium to long-term biomarker of exposure to petroleum-contaminated water bodies.

Persistent hyperplastic primary vitreous with retinal tumor in tuberous sclerosis: report of a case including tumoral immunohistochemistry and cytogenetic analyses.

OBJECTIVE: The authors describe an ocular lesion combining the characteristics of persistent hyperplastic primary vitreous (PHPV) and a retinal tumor in an infant with tuberous sclerosis complex (TSC). STUDY DESIGN: Case report. METHODS: Immunohistochemistry and cytogenetic studies were performed on TSC cells from an intraocular tumor in a 6-week-old infant. RESULTS: Histopathologic examination showed a thick fibrovascular membrane between the aspect of the lens and the astrocytic component of the mass. Glial fibrillary acidic protein (GFAP) showed a variable intracytoplasmic reaction in the astrocytic proliferation, involving approximately 50% of the cells. Tissue culture studies showed a fairly rapid proliferation of fusiform cells, consistent with bipolar astrocytic cells. Cytogenetic studies showed one abnormal clone consisting of three hyperdiploid cells with a loss of chromosome 9 and a gain of chromosomes 6 and 12. CONCLUSION: The atypical localization of the retinal tumor could be explained by the fact that it was trapped during its proliferation by the retinal detachment associated with the PHPV.

Corneal endothelial cell density in glaucoma.

PURPOSE: We studied corneal endothelial cell density in patients with glaucoma. METHODS: One hundred two patients with glaucoma were compared with 52 patients without glaucoma of the same age group. Exclusion criteria included history of either corneal disease, ocular inflammation, trauma, or surgery other than peripheral iridectomy. The following data were extracted from the patient files: glaucoma type and duration, laser treatments, glaucoma medications, and documented intraocular pressure (IOP) measurements. Specular microscopies were performed on central corneas, endothelial images were analyzed by computerized planimetry, and cell counts were calculated. RESULTS: Corneal endothelial cell counts were significantly lower in patients with glaucoma (2,154 +/- 419 cells/mm2) than in controls (2,560 +/- 360 cells/mm2; t test, p < 0.0001). In the glaucoma group, cell counts were inversely proportional to the means of IOPs. Patients receiving three or four glaucoma medications had lower cell counts than those receiving one or two medications. Cell counts were significantly lower both in primary angle-closure glaucoma and in primary open-angle glaucoma. CONCLUSION: This study suggests that patients with glaucoma may have lower corneal endothelial cell density than those without glaucoma of the same age group. The proposed mechanisms are direct damage from IOP, congenital alteration of the corneal endothelium in patients with glaucoma, glaucoma medication toxicity, or a combination of these.

Intraindividual reproducibility of postprandial and orthostatic blood pressure changes in older nursing-home patients: relationship with chronic use of cardiovascular medications.

OBJECTIVES: Although postprandial and orthostatic hypotension are commonly observed in nursing home residents, their reproducibility, relationship to each other, and association with chronic use of cardiovascular medications are poorly understood. DESIGN: We examined blood pressure (BP) and heart rate (HR) before and after postural change, and before and after a 419-kcal meal in 22 nursing home residents (mean age 89 +/- 5 (SD) years), each on two occasions, to determine reproducibility changes. These studies were repeated in 17 residents, with and without previous administration of cardiovascular medications, in random order. SETTING: Hebrew Rehabilitation Center for the Aged, an academic long-term care facility. RESULTS: Systolic BP declined an average (+/- SE) of 16 +/- 4 mm Hg and 12 +/- 4 mm Hg during the first and second meal studies, respectively. Mean intra-class correlation of postprandial systolic BP values during the two studies was 0.88 (95% CI 0.85-0.97). Systolic BP increased significantly during the first posture test to a maximum of 8 +/- 6 mm Hg at 6 minutes. There was no significant difference over time in postural systolic BP between the two tests. Repeated postural studies showed a mean intra-class correlation of 0.72 (95% CI 0.62-0.92) for changes in systolic BP. Cardiovascular medications had no additional effect on postprandial or orthostatic BP and HR changes. During the first studies, 10 subjects had postprandial hypotension, and three subjects had orthostatic hypotension, but only two of 22 subjects had both. CONCLUSIONS: Patterns of systolic BP response to meals or postural change are reproducible. BP responses to meals and postural change seem to be unaffected by potentially hypotensive medications in chronic users. Postprandial hypotension is distinct from orthostatic hypotension, occurring more commonly than orthostatic hypotension and infrequently together in the same patients.