Keerthi S. Guruge

Perfluorinated organic compounds in human blood serum and seminal plasma: a study of urban and rural tea worker populations in Sri Lanka

Concentrations and accumulation of 13 fluorinated organic compounds (FOCs) in human sera and seminal plasma were measured in an Asian developing country, Sri Lanka. Six of the FOCs, PFOS (perfluorooctanesulfonate), PFHS (perfluorohexanesulfonate), PFUnA (perfluoroundecanoic acid), PFDA (perfluorodecanoic acid), PFNA (perfluorononanoic acid) and PFOA (perfluorooctanoic acid), were detected in all of the sera samples. Measurable quantities of two main perfluorosulfonates, PFOS and PFHS, were found in all seminal plasma samples. The detection frequency of the predominant perfluoroalkylcarboxylate, PFOA, in seminal plasma was >70%. Accumulation of PFOS in sera was significantly positively correlated with PFOA, PFHS and PFNA. Positive linear regressions were also found between PFNA and PFUnA and PFNA and PFDA suggesting that these compounds may have a similar origin of exposure and accumulation. Significantly positive associations were observed for partitioning of both PFOS and PFNA between sera and seminal plasma. The accumulation of FOCs was not significantly different in sera from Colombo (urban population) and Talawakele (rural conventional tea workers). However, the Haldummulla population (rural organic tea workers) had relatively lower exposure to FOCs compared to the other two groups, urban and rural conventional tea workers. Concentrations of FOCs in Sri Lanka were similar to those reported for industrialized countries suggesting that human exposure to such chemicals is widespread even in developing countries. The novel finding of FOCs in human seminal plasma implies that further studies are needed to determine whether long-term exposure in humans can result in reproductive impairments.

Differential expression of chicken hepatic genes responsive to PFOA and PFOS

The effects of PFOS and PFOA on the gene expression patterns of chickens that were exposed to either PFOS or PFOA at low doses were investigated with the use of microarray techniques. Twelve Genechip Chicken Genome Arrays were used to study hepatic gene expression in 6-week-old chickens (Gallus gallus) that were exposed to either PFOA (0.1, 0.5, or 5mg/mL), PFOS (0.02 or 0.1mg/mL), or a saline vehicle control (0.9% NaCl in Milli-Q water) via subcutaneous implantation of a 2mL osmotic pump for 4 weeks or for 4 weeks with a further 4 weeks of depuration. Over 240 and 480 genes were significantly affected by PFOS after 4 weeks of exposure and after 4 weeks of exposure with a further 4 weeks of depuration, respectively and over 290 and 320 genes were significantly affected by PFOA, correspondingly. For PFOS, the genes that were affected after 4 weeks of exposure were mainly related to the transport of electrons and oxygen, and the metabolism of lipids and fatty acids; while the genes that were affected after 4 weeks of exposure with a further 4 weeks of depuration were mainly related to the transport of electrons and ions, and protein amino acid phosphorylation and proteolysis. For PFOA, the genes that were affected after 4 weeks of exposure were related to the transport of ions, lipids, and electrons and cytochromes; while the genes that were affected after 4 weeks of exposure with a further 4 weeks of depuration were related to protein amino acid phosphorylation and proteolysis, the transport of ions, and the metabolism of fatty acids and lipids. The results also showed that the gene expression patterns between chickens that were treated with PFOS and those that were treated with PFOA were different, which points to the importance of the separate evaluation of the toxicities of PFOS and PFOA. Specifically, the gene expressions of CYP8B and NOV were studied.

Evidence for the involvement of xenobiotic-responsive nuclear receptors in transcriptional effects upon perfluoroalkyl acid exposure in diverse species

Humans and ecological species have been found to have detectable body burdens of a number of perfluorinated alkyl acids (PFAA) including perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS). In mouse and rat liver these compounds elicit transcriptional and phenotypic effects similar to peroxisome proliferator chemicals (PPC) that work through the nuclear receptor peroxisome proliferator-activated receptor alpha (PPAR alpha). Recent studies indicate that along with PPAR alpha other nuclear receptors are required for transcriptional changes in the mouse liver after PFOA exposure including the constitutive activated receptor (CAR) and pregnane X receptor (PXR) that regulate xenobiotic metabolizing enzymes (XME). To determine the potential role of CAR/PXR in mediating effects of PFAAs in rat liver, we performed a meta-analysis of transcript profiles from published studies in which rats were exposed to PFOA or PFOS. We compared the profiles to those produced by exposure to prototypical activators of CAR, (phenobarbital (PB)), PXR (pregnenolone 16 alpha-carbonitrile (PCN)), or PPAR alpha (WY-14,643 (WY)). As expected, PFOA and PFOS elicited transcript profile signatures that included many known PPAR alpha target genes. Numerous XME genes were also altered by PFOA and PFOS but not WY. These genes exhibited expression changes shared with PB or PCN. Reexamination of the transcript profiles from the livers of chicken or fish exposed to PFAAs indicated that PPAR alpha, CAR, and PXR orthologs were not activated. Our results indicate that PFAAs under these experimental conditions activate PPAR alpha, CAR, and PXR in rats but not chicken and fish. Lastly, we discuss evidence that human populations with greater CAR expression have lower body burdens of PFAAs.

Species-specific concentrations of perfluoroalkyl contaminants in farm and pet animals in Japan

The persistent metabolites of perfluorinated compounds (PFCs) which have been detected in the tissues of both humans and wildlife, and human contamination by PFCs suggest differences in the exposure patterns to these compounds. However, studies focused on identifying human exposure pathways to PFCs are scarce. To provide a preliminary assessment of PFCs in farm animals such as chicken, cattle, pigs, goats and horses, blood and liver samples were collected from various regions in Japan. Additionally, dog sera samples representing pet animals were also employed for analysis. Perfluorooctane sulfonate (PFOS) was the most prominent contaminant found in farm and pet animals, with mean sera PFOS concentrations (in decreasing order) of: chicken (5.8 ng/ml)>cattle (3.0 ng/ml)>goat (2.4 ng/ml)>horse (0.71 ng/ml)>pig (0.37 ng/ml). Chicken livers (67 ng/g) contained the highest mean PFOS concentration among the farm animals, followed by those of pigs (54 ng/g) and cattle (34 ng/g). In comparison to PFOS levels in farm animals, the detected levels of other PFCs were not significant. The high levels of PFOS found in cattle fetal livers suggest that PFOS crosses the placental barrier to enter fetal circulation. The consumption of chicken by humans might produce higher PFOS exposure in humans compared to that in farm animals; however, the current levels of PFOS in farm animals in Japan were lower than those reported in fish and wild animals. Elevated concentrations of both PFOS (25 ng/ml) and perfluorohexane sulfonate (PFHxS; 10 ng/ml) were found in dog sera, indicating that further studies are needed to identify PFC sources in the human environment.

An analytical method for the determination of perfluorinated compounds in whole blood using acetonitrile and solid phase extraction methods

A method for the analysis of perfluorinated compounds (perfluoroalkyl sulfonates: C4, C6, C8, C10; perfluoroalkyl sulfinates: C6, C8, C10; perfluorooctanesulfonamide, N-ethyl perfluorooctanesulfonamide, N-ethyl perfluorooctanesulfonamidoacetate, perfluorocarboxylates: C4-C14; fluorotelomer carboxylate (7:3, 8:2) in whole blood using acetonitrile and OASIS WAX solid phase extraction (SPE) cartridge was developed. Separation of target compounds in two HPLC columns (ion exchange JJ50-2D and C18 Betasil columns) was examined. Matrix recoveries of the developed methods ranged from 70% to 120%. Separation of possible inferences such as taurodeoxycholic acid (TDC) was accomplished using an ion exchange JJ50-2D column, and this separation was validated using whole blood of different animals.

Depuration kinetics and tissue disposition of PFOA and PFOS in white leghorn chickens (Gallus gallus) administered by subcutaneous implantation.

Elimination kinetics and tissue disposition of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) in male chickens (Gallus gallus) was determined following exposure by subcutaneous implantation. Chickens were exposed to two levels of PFOA or PFOS for 4wk and then allowed to depurate for an additional 4wk. These exposures did not cause any statistically significant changes in body index, clinical biochemistry or histology among treatments relative to the controls (p>0.05), except that concentrations of total cholesterol and phospholipids were less in chickens exposed to PFOS. The elimination rate constant for PFOA (0.150+/-0.010d(-1)) was approximately six-fold greater than that of PFOS (0.023+/-0.004d(-1)). The greatest concentrations of PFOA and PFOS were found in kidney and liver, respectively. The organ to blood ratio of PFOS concentration was increased after the whole experiment, indicating the importance of organ partitioning of PFOS in elimination kinetics. The depuration half-life of PFOA (t(1/2)=4.6d) and PFOS (t(1/2)=125d) in chickens was calculated.

Fate of maize intrinsic and recombinant genes in calves fed genetically modified maize Bt11.

The presence of maize intrinsic and recombinant cry1Ab genes in the gastrointestinal (GI) contents, peripheral blood mononuclear cells (PBMC), and visceral organs of calves fed genetically modified Bt11 maize was examined by PCR in a subchronic 90-day performance study. Samples were collected from six Japanese Black/Holstein calves fed Bt11 maize and from six calves fed non-Bt maize. Fragments of maize zein (Ze1), invertase, chloroplast, and cry1Ab were detected inconsistently in the rumen fluid and rectal contents 5 and 18 h after feeding. The chloroplast DNA fragments of ribulose-1,5-bisphosphate carboxylase/oxygenase and tRNA were detected inconsistently in the PBMC, the visceral organs, and the longissimus muscle, while the cry1Ab gene was never detected in PBMC or in the visceral organs. These results suggest that feed-derived maize DNA was mostly degraded in the GI tract but that fragmented DNA was detectable in the GI contents as a possible source of transfer to calf tissues. These results also suggest that the recombinant cry1Ab genes were not transferred to the PBMC and tissues of calves fed Bt11 maize.

A review of the occurrence of pharmaceuticals and personal care products in Indian water bodies

Little information exists on the occurrence and the ultimate fate of pharmaceuticals in the water bodies in India despite being one of the world leaders in pharmaceutical production and consumption. This paper has reviewed 19 published reports of pharmaceutical occurrence in the aquatic environment in India [conventional activated sludge wastewater treatment plants (WTPs), hospital WTPs, rivers, and groundwater]. Carbamazepine (antipsychoactive), atenolol (antihypertensive), triclocarban and triclosan (antimicrobials), trimethoprim and sulfamethoxazole (antibacterials), ibuprofen and acetaminophen (analgesics), and caffeine (stimulant) are the most commonly detected at higher concentrations in Indian WTPs that treat predominantly the domestic sewage. The concentration of ciprofloxacin, sulfamethoxazole, amoxicillin, norfloxacin, and ofloxacin in Indian WTPs were up to 40 times higher than that in other countries in Europe, Australia, Asia, and North America. A very few studies in Indian rivers reported the presence of ciprofloxacin, enoxacin, ketoprofen, erythromycin, naproxen, ibuprofen, diclofenac and enrofloxacin. Similar compounds were reported in rivers in China, indicating a similar usage pattern in both of these developing countries. In a study reported from an open well in southern India, the groundwater showed the presence of cetirizine, ciprofloxacin, enoxacin, citalopram and terbinafine, which was close to a WTP receiving effluents from pharmaceutical production.

Comparison of total fluorine, extractable organic fluorine and perfluorinated compounds in the blood of wild and pefluorooctanoate (PFOA)-exposed rats: Evidence for the presence of other organofluorine compounds

The widespread occurrence and environmental persistence of perfluorinated compounds (PFCs) received worldwide attention recently. Exhaustive analysis of all fluorinated compounds in an environmental sample can be daunting because of the constraints in the availability of analytical standards and extraction methods. Combustion ion chromatographic technique for trace fluorine analysis was used to assess the concentrations of known PFCs (e.g., PFOS, PFOA) and total fluorine (TF) in the blood of wild rats collected from Japan. The technique was further validated using tissues from PFOA-exposed rats. Six PFCs (PFOS, PFOSA, PFUnDA, PFDA, PFNA, and PFOA) were detected in all of the wild rat blood samples. Concentrations of extractable organic fluorine (EOF) in fraction 1 (Fr1; MTBE extraction) of wild rats ranged 60.9-134 ng F mL(-1), while those in fraction 2 (Fr2; hexane) were below LOQ (32 ng F mL(-1)); TF concentrations in the blood of wild rats ranged from 59.9-192 ng F mL(-1). The contribution of known PFCs in EOF-Fr1 (MTBE) varied from 9% to 89% (56% on average), and known PFC concentrations in TF content were less than 25%. In contrast, TF concentrations in the blood of PFOA-exposed rats ranged from 46900 to 111,000 ng F mL(-1), with PFOA contributing over 90% of TF. A comparison of results from the samples analyzed in this study and the literature revealed three distinct groups with PFOA/known PFC and TF levels (i.e., wild rats and general population, occupationally exposed workers, and PFOA-exposed laboratory rats). The mass balance analysis of the different forms of fluorine in blood suggested the presence of other forms of organic fluorine in addition to known PFCs.

Impact of wastewater from different sources on the prevalence of antimicrobial-resistant Escherichia coli in sewage treatment plants in South India.

The sewage treatment plant (STP) is one of the most important interfaces between the human population and the aquatic environment, leading to contamination of the latter by antimicrobial-resistant bacteria. To identify factors affecting the prevalence of antimicrobial-resistant bacteria, water samples were collected from three different STPs in South India. STP1 exclusively treats sewage generated by a domestic population. STP2 predominantly treats sewage generated by a domestic population with a mix of hospital effluent. STP3 treats effluents generated exclusively by a hospital. The water samples were collected between three intermediate treatment steps including equalization, aeration, and clarification, in addition to the outlet to assess the removal rates of bacteria as the effluent passed through the treatment plant. The samples were collected in three different seasons to study the effect of seasonal variation. Escherichia coli isolated from the water samples were tested for susceptibility to 12 antimicrobials. The results of logistic regression analysis suggest that the hospital wastewater inflow significantly increased the prevalence of antimicrobial-resistant E. coli, whereas the treatment processes and sampling seasons did not affect the prevalence of these isolates. A bias in the genotype distribution of E. coli was observed among the isolates obtained from STP3. In conclusion, hospital wastewaters should be carefully treated to prevent the contamination of Indian environment with antimicrobial-resistant bacteria.