Natural attenuation of endocrine-disrupting estrogens in an ecologically engineered treatment system (EETS) designed with floating, submerged and emergent macrophytes

Natural attenuation of estrogenic endocrine disrupting compounds (EDCs) such as estriol (E3, natural) and 17α-ethinylestradiol (EE2, synthetic) were evaluated in a designed ecologically engineered treatment system (EETS) along with domestic sewage. These two estrogens are the major contaminants of sewage and found to cause adverse effects on the endocrine system of humans and animals when exposed even in nanogram concentrations. The EETS consisted of three tanks containing diverse biota, viz., aquatic macrophytes, submerged plants, emergent plants, algae and bacteria present in the system mimic the natural cleansing functions of wetlands and help in the treatment of pollutants present in wastewater. During operation, 22 μg/l of E3 and EE2 were separately fed for 10 days each and operated in continuous mode (20 l/day). The floating macrophytes system (Tank 1) was more effective in removing estrogens [E3 - 61.77% (13.59 μg/l); EE2 - 69.09% (15.20 μg/l)] compared to the submerged-emergent macrophytes-based integrated system (Tank 2) [E3 - 16.58% (3.65 μg/l); EE2 - 18.52% (4.08 μg/l)] and submerged-rooted microphytes system (Tank 3) [E3 - 15.20%, (3.35 μg/l); EE2 - 7.72%, (1.70 μg/l)]. On the whole, EETS can effectively treat EDCs [E3 93.56% (20.59 μg/l); EE2 95.34% (20.97 μg/l)]. Removal of COD (68.06%), nitrates (60.02%) and turbidity (83.43%) was also observed simultaneously during EETS operation. The designed EETS is ecologically complex and mechanically simple and has very low energy consumption and function based on a natural cleansing mechanism (attenuation) with esthetic value.     

The negative impact of the environment on methylation/epigenetic marking in gametes and embryos: A plea for action to protect the fertility of future generations

Life expectancy has increased since World War II, and this may be attributed to several aspects of modern lifestyles. However, now we are faced with a downturn, which seems to be the result of environmental issues. This paradigm is paralleled with reduced human fertility, decreased sperm quality, increased premature ovarian failure, and diminished ovarian reserve syndromes. Endocrine disruptor chemicals and other toxic chemicals, herbicides, pesticides, plasticizers, to mention a few, are a rising concern in today's environment. Some of these are commonly used in the domestic setting: cleaning material and cosmetics and they have a known impact on epigenesis and imprinting via perturbation of methylation processes. Pollution from polyaromatic hydrocarbons, particulate matter <10 and <2.5 μm, and ozone released into the air, all affect fertility. Poor food processing management is a source of DNA adduct formation, which impairs the quality of gametes. An important concern is the nanoparticles that are present in food and are thought to induce oxidative stress. Now is the time to take a step backward. Global management of the environment and food production is required urgently to protect the fertility of future generations.     

Toxicity monitoring of endocrine disrupting chemicals (EDCs) using freeze-dried recombinant bioluminescent bacteria

Five different freeze-dried recombinant bioluminescent bacteria were used for the detection of cellular stresses caused by endocrine disrupting chemicals. These strains were DPD2794 (recA::luxCDABE), which is sensitive to DNA damage, DPD2540 (fabA::luxCDABE), sensitive to cellular membrane damage, DPD2511 (katG::luxCDABE), sensitive to oxidative damage, and TV1061 (grpE::luxCDABE), sensitive to protein damage. GC2, which emits bioluminescence constitutively, was also used in this study. The toxicity of several chemicals was determined on the first four freeze-dried bacteria, while nonspecific cellular stresses were measured using GC2. Damage caused by known endocrine disrupting chemicals, such as nonyl phenol, bisphenol A, and styrene, was detected and classified according to toxicity mode, while others, such as phathalate and DDT, were not detected with the bacteria. These results suggest that endocrine disrupting chemicals are toxic in bacteria, and do not act via an estrogenic effect, and that toxicity monitoring and classification of some endocrine disrupting chemicals may be possible in the field using these freeze-dried recombinant bioluminescent bacteria.     

The environmental endocrine disruptor, bisphenol A, affects germination, elicits stress response and alters steroid hormone production in kiwifruit pollen

In vitro toxicity of the endocrine disruptor bisphenol A (BPA) to pollen, the male haploid generation of higher plants, was studied. BPA caused significant inhibition of both tube emergence and elongation of kiwifruit pollen in a dose-dependent manner, beginning at 10 mg · l(-1); morphological changes to tubes were also detected. Despite strong inhibition of pollen tube production and growth, a large percentage of treated cells remained viable. Immunoblotting experiments indicated that levels of BiP and 14-3-3, which are proteins involved in stress response, substantially increased in BPA-treated pollen compared to controls. The increases were dose-dependent in the range 10-50 mg · l(-1) BPA, i.e. even when germination ability was completely blocked. Steroid hormones (17 β-estradiol, progesterone and testosterone) were detected in kiwifruit pollen, and their levels increased during germination in basal medium. In a BPA treatment of 30 mg · l(-1), larger increases in both estrogen and testosterone concentrations were detected, in particular, a six-fold increase of 17 β-estradiol over control concentration (30 min). The increased hormone levels were maintained for at least the 90 min incubation. Increasing concentrations of exogenous testosterone and 17 β-estradiol increasingly inhibited pollen tube emergence and elongation. Current data for BPA-exposed kiwifruit pollen suggest a toxicity mechanism that is at least in part based on a dramatic imbalance of steroid hormone production during tube organisation, emergence and elongation. It may be concluded that BPA, a widespread environmental contaminant, can cause serious adverse effects to essential pollen functions. On a broader scale, this chemical poses a potential risk to the reproductive success of higher plants.     

Characterization and expression profile of Vitellogenin gene from Scylla paramamosain

The full-length (7816 bp) cDNA of Vitellogenin (Vtg) encoding 2560 aa with an estimated molecular mass of 287.743 kDa was cloned from the green mud crab Scylla paramamosain. Semi-quantitative PCR (sq-PCR) revealed a specific expression pattern of Sp-vtg gene in ovaries and hepatopancreas. With the development of ovaries, the expression level of Sp-vtg gene showed an increasing trend both in ovaries and hepatopancreas, and the expression level of Sp-vtg gene in hepatopancreas and ovary was stable after stage IV. By in situ hybridization, the positive signals of Sp-vtg gene were detected in the cytoplasm of oocytes in stage I, in the follicle cell and the surrounding of the nucleus in stage III, and in the nucleus in stage V. Furthermore, the signals become stronger with the later development stages of ovary. Moreover, in situ hybridization analysis revealed that positive signals of Sp-vtg gene are present in the hepatopancreatic tubule, and the signals increase during the development, becoming the strongest in stage V. Our results indicate that both ovaries and hepatopancreas are sites of Vitellogenin gene synthesis in S. paramamosain.     

Enhanced expression of laccase during the degradation of endocrine disrupting chemicals in Trametes versicolor

A putative laccase cDNA from a white-rot basidiomycete, Trametes versicolor, that consisted of 1,769 nucleotides was cloned using the rapid amplification of cDNA ends (RACE)-PCR method. The deduced amino acid sequence had 4 putative copper binding regions, which are common to fungal laccases. In addition, the sequence was 57 approximately 97 % homologous to sequences of other T. versicolor laccases. Additionally, the expression of laccase and manganese peroxidase in this fungus were both greatly increased under degrading conditions for bisphenol A, nonylphenol and two phthalic esters (benzylbutylphthalate and diethylphthalate), all of which are reportedly endocrine disrupting chemicals (EDCs). Furthermore, the estrogenic activities of the EDCs also decreased rapidly during incubation when examined in a two-hybrid yeast system. Finally, kojic acid inhibited the removal of estrogenic activities generated by bisphenol A and nonylphenol, which confirmed that laccase was involved in the degradation of EDCs in T. versicolor.