The present study aimed to use primary liver cell culture derived from the orange-spotted grouper, Epinephelus coioides, to assess the toxic effects of nonylphenol (NP) on the hepatocyte viability and the liver antioxidant system. E. coioides was selected due to its commercial importance. NP was used in this study because of its high potential of producing oxidative stress due to increased reactive oxygen species (ROS). A liver of E. coioides was digested with PBS containing 0.1% collagenase IV. The digested cells were moved to Leibovitz L-15 culture medium with 20% fetal bovine serum (FBS), 100IUmL−1 penicillin, 100μgmL−1 streptomycin. Aliquots of cell suspension were seeded as a monolayer into sterile 25cm2 tissue culture flasks and incubated at 30°C for 14days. The medium, containing non-attached cells, was removed after 24 to 48h and a new medium was added. The IC50 of 10−4molL−1 was determined for nonylphenol using MTT assay. Cells were then incubated with L-15 medium containing 10−5, 2×10−5, 3×10−5molL−1 of NP and samples were taken after 6, 12 and 24h of incubation for analysis of LPO, SOD, CAT, GPx, LDH, AST, ALT, and ALP. Based on the results, the lowest concentration of NP was not markedly cytotoxic to primary hepatocytes and the cell sensitivity to NP increased dose-dependently. The activities of SOD, CAT and GPx decreased significantly, while activities of LPO, LDH, AST, ALT and ALP, increased significantly in a dose-related pattern in NP-treated cells. In conclusion, this study revealed that NP could induce the oxidative stress in cultivated hepatocytes of E. coioides during a short-term exposure. NP toxicity is mainly due to the induction of the reactive oxygen species (ROS), which lead to cell membrane disruption, damage of cellular metabolism, and interference with cellular macromolecules.

The relationship between chemical exposure and disease outbreak in fish has not been fully defined due to the limitations of experimental systems (model fish and pathogens). Therefore, we constructed a system using the Japanese flounder, Paralichthys olivaceus, and viral haemorrhagic septicemia virus (VHSV), and evaluated it by heavy oil (HO) exposure. The fish were exposed to HO at 0.3, 0.03, 0.003, and 0g/L following VHSV infection at doses of 10².⁵ or 10³.⁵ tissue culture infectious dose (TCID)₅₀/fish. As a result, groups given the dual stressors showed more than 90% mortality. Although VHSV infection at 10².⁵ and 10³.⁵ TCID₅₀/fish without HO exposure also induced high mortality, at 68.8% and 81.3%, respectively, HO exposure induced faster and higher mortality in the virus carrier fish, indicating that chemical stressors raise the risk of disease outbreak in fish. The experimental system established in this study could be useful for chemical risk assessment.

In this paper, we are focusing on study of the impact of sterilization on culture establishment of indigenous Musa variety to restore plant diversity and its importance in human life. Biodiversity supports biological system profitability where every species has a significant task to carry out. Biodiversity of plants guarantees an asset for new nourishment harvests and prescriptions. The biodiversity conservation can start from in situ and ex situ techniques. In situ strategies secure the plants in their common natural surroundings, where as ex situ techniques include safeguarding and upkeep of plant species through plant tissue culture technique. Re-establishing Indigenous plant territory is fundamental for conservation of biodiversity. Indigenous plants are those that present normally in a place where they developed. They are the biological premise whereupon the life of animals and human depends. Traditional medicines from plants are significant part of Indian culture. Banana cultivars nowadays are triploid (AAA, AAB, ABB) cross breeds of two species, Musa acuminate Colla (AA) and Musa balbisiana Colla (BB), which separately contribute the A and B genomes. The M. balbisiana which is normally present in North-Eastern belt of India has high nutrients and resistance to biotic and abiotic stresses. Nanjangud rasabale has been found in parts of Mysuru district, known for its stand-out smell, flavor, taste, and time span of sensible convenience anyway incredibly viewed as a jeopardized grouping; red bananas are phenomenal anyway concerning supplement C and potassium, they are better than yellow ones; Meitei Hei is acclaimed for its enjoyableness and incredible alluring quality. It is a cold permissive grouping thriving incredible even at 12–15 °C. The accomplishment of plant tissue culture convention relies upon explant sterilization procedure. Determination of sterilant and time span of introduction are additionally basic on the grounds that the living material should not lose their natural action and just contaminants should be wiped out during cleansing. Surface sanitization of explant is a procedure which includes the drenching of explants into suitable convergence of compound sterilants for a predetermined time bringing about the foundation of a contamination-free culture.

Understanding plant behaviour in polluted soils is critical for the sustainable remediation of metal-polluted sites including abandoned mines. Post-operational and abandoned metal mines particularly in semi-arid and arid zones are one of the major sources of pollution by soil erosion or plant hyperaccumulation bringing ecological impacts. We have selected from the literature 157 species belonging to 50 families to present a global overview of ‘plants under action’ against heavy metal pollution. Generally, all species of plants that are drought, salt and metal tolerant are candidates of interest to deal with harsh environmental conditions, particularly at semi-arid and arid mine sites. Pioneer metallophytes namely Atriplex nummularia, Atriplex semibaccata, Salsola kali, Phragmites australis and Medicago sativa, representing the taxonomic orders Caryophyllales, Poales and Fabales are evaluated in terms of phytoremediation in this review. Phytoremediation processes, microbial and algal bioremediation, the use and implication of tissue culture and biotechnology are critically examined. Overall, an integration of available remediation plant-based technologies, referred to here as ‘integrated remediation technology,’ is proposed to be one of the possible ways ahead to effectively address problems of toxic heavy metal pollution. Graphical abstract Integrated remediation technology (IRT) in metal-contaminated semi-arid and arid conditions. The hexagonal red line represents an IRT concept based on remediation decisions by combination of plants and microbial processes.

The problems that most affect the quality of the waters of rivers and lakes are associated with the discharges performed in these environments, mainly industrial and domestic effluents inappropriately treated or untreated. The comet assay is a sensitive tool and is recommended for studies of environmental biomonitoring, which aim to determine the genotoxicity potential of water pollutants. This study aimed to assess the genotoxic potential of the Ribeirão Tatu waters, region of Limeira, São Paulo (SP), by the comet assay with mammalian cells (hepatoma tissue culture (HTC)). Water samples were collected along the Ribeirão Tatu at three distinct periods: November 2008, February 2009 and August 2009, and five collection sites were established: P1, source of the stream; P2, site located downstream the urban perimeter of the municipality of Cordeirópolis and after receiving the pollution load of this city; P3, collection site located upstream the urban perimeter of the city of Limeira; P4, urban area of Limeira; and P5, rural area of Limeira, downstream the discharges of the city sewage. The results showed that for the November 2008 collection, there was no water sample-induced genotoxicity; for the February 2009 collection, the sites P1 and P2 were statistically significant in relation to the negative control (NC), and for the August 2009 collection, the site P5 was statistically significant. These results could be explained by the content of different metals during the different seasons that are under the influence of domestic, industrial and agricultural effluents and also due to the seasonality, since the water samples collected in the period of heavy rain (February 2009) presented a higher genotoxicity possibly due to the entrainment of contaminants into the bed of the stream promoted by the outflow of rainwaters. The comet assay showed to be a useful and sensitive tool in the evaluation of hydric resources impacted by pollutants of diverse origins, and a constant monitoring should be done in order to verify the influence of different factors (season, amount of contaminants) in the water quality.

Populus species are very effective in remediation of contaminants. Polybrominated diphenyl ethers (PBDEs) are commonly used as flame retardants and are known to be persistent environmental pollutants. Numerous studies have shown that PBDEs are rising in human tissues and biota. 4-Monobrominated diphenyl ether (4-BDE), one of the less brominated PBDEs, was served as a model compound for biodegradation of lower brominated congeners. The present study was designed to clarify the effects of 4-BDE stress on morphological, physiological, and biochemical impacts of Populus tomentosa Carr in a tissue culture condition. Different concentrations of 4-BDE (3 and 30 mg L⁻¹) were supplied alone or together with 0.5 mg L⁻¹ IBA in tissue culture media. With the concentration increased, 4-BDE caused negative effects on the microscopic structure of roots, stem, and leaves. The leaf color became shallow in low concentration of 4-BDE treatments and appeared albinism with 4-BDE concentration increased. The chlorophyll content and the leaf mass per area of albino leaves reduced significantly. 4-BDE also caused positive effects on the adventitious root differentiation and the biomass below 30 mg L⁻¹. With the 4-BDE treatment time increased (23, 47, and 58 days), the peroxidase (POD) activity displayed the decreasing trend. The proline content decreased first and then increased. Exposure to 4-BDE induced the malondialdehyde (MDA) to increase in leaves. Application of 4-BDE affected the endogenous hormone levels of cuttings in their adventitious roots inducing media. Below 0.3 mg L⁻¹, 4-BDE caused the faint expression of auxin-sensitive DR5::GUS reporter gene in Arabidopsis thaliana. Additionally, P. tomentosa Carr exhibited the better tolerance against 4-BDE in the range of less than 30 mg L⁻¹.

BACKGROUND, SCOPE, AND AIMS: Antibacterial fluoroquinolones (FQs) are third-generation antibiotics that are commonly used as therapeutic treatments of respiratory and urinary tract infections. They are used far less in intensively farmed animal production systems, though their use may be permitted in the veterinary treatments of flocks or in medicated feeds. When used, only a fraction of ingested parent FQ actually reaches the in vivo target site of infection, while the remainder is excreted as the parent FQ and its metabolized products. In many species’ metabolism, enrofloxacin (EF) is converted into ciprofloxacin (CF) while both FQs are classified as parent FQs in human treatments. It is therefore likely that both FQs and their metabolic products will contribute to a common pool of metabolites in biological wastes. Wastes from intensive farming practices are either directly applied to agricultural land without treatment or may be temporarily stored prior to disposal. However, human waste is treated in sewage treatment plants (STPs) where it is converted into biosolids. In the storage or treatment process of STPs, FQs and their in vivo metabolites are further converted into other environmental metabolites (FQEMs) by ex vivo physicochemical processes that act and interact to produce complex mixtures of FQEMs, some of which have antibacterial-like activities. Biosolids are then often applied to agricultural land as a fertilizer amendment where FQs and FQEMs can be further converted into additional FQEMs by soil processes. It is therefore likely that FQ-contaminated biowaste-treated soils will contain complex mixtures of FQEMs, some of which may have antibacterial-like activities that may be expressed on bacteria endemic to the receiving agricultural soil environment. Concern has arisen in the scientific and in the general community that repeated use of FQ-contaminated biowaste as fertilizer amendments of nutrient-impoverished agricultural land may create a selective environment in which FQ-resistant bacteria might grow. The likelihood of this happening will depend, to some extent, on whether bioactive FQEMs are first synthesized from the parent FQs by the action and interaction of in vivo and ex vivo processes producing bioactive FQEMs in biowastes and biosolids. The postulated creation of a selective environment will also depend, in part, on whether such bioactive FQEMs are biologically available to bacteria, which may, in turn, be influenced by soil type, amendment regime, and the persistence of the bioactive FQEMs. Additionally, soil bacteria and soil processes may be affected in different ways or extents by bioactive FQEMs that could possibly act additively or synergistically at ecological targets in these non-target bacteria. This is an important consideration, since, while parent FQs have well-defined ecological targets (DNA gyrase and topoisomerase IV) and modes of bactericidal action, the FQEMs and their possible modes of action on the many different species of soil bacteria is less well studied. It is therefore understandable that there is a lack of conclusive evidence directly attributing biosolid usage to any increase in FQ-resistant bacteria detected in biowaste-amended agricultural soil. However, a lack of evidence may simply imply that a causal relationship between biosolid usage programs and any detection of low levels of FQ-resistant bacteria in soils has yet to be established, rather than an assumption of no relationship whatsoever. Based on results presented in this paper, the precautionary principle should be applied in the usage of FQ-contaminated biosolids as fertilizer amendments of agricultural land. The aim of this research was to test whether any bioactive FQEMs of EF could be synthesized by aerobic fermentation processes using Mycobacterium gilvum (American Tissue Culture Collection) and a mixed culture of microorganisms derived from an agricultural soil. High-performance thin-layer chromatography (HPTLC) and bioautography were tested as screening techniques in the detection and analysis of bioactive FQEMs. MATERIALS AND METHODS: FQEMs derived from M. gilvum and mixed (soil) culture aerobic ferments were fractionated using preparative HPTLC. A standard strain of Escherichia coli was then used as the reporter organism in a bioautography assay in the detection of bioactive-FQEMs on a mid-section of the HPTLC plate. Plate sections were reassembled, and a photograph was taken under low-intensity ultraviolet (UV) light to reveal regions that contained analytes that had UV chromophores and antibacterial-like activities. RESULTS AND DISCUSSION: Many fractionated FQEMs displayed antibacterial-like activity while bound to silica gel HPTLC plates. These results also provide evidence that sufficient quantities of biologically active FQEMs were biologically available from a silica gel surface to prevent the adherent growth of E. coli. Six to seven FQEMs derived from EF using aerobic fermentation processes had antibacterial-like activities, while two FQEMs were also detectable using UV light. Furthermore, similar banding patterns of antibacterial-like activity were observed in both the monoculture (M. gilvum) and mixed culture bioautography assays, indicating that similar processes operated in both aerobic fermentations, either producing similar biologically active FQEMs or biologically active FQEMs that had similar physicochemical properties in both ferments. The simplest explanation for these findings is that the tested agricultural soil also contained mycobacteria that metabolized EF in a similar way to the purchased standard monoculture M. gilvum. Additionally, the marked contrast between the bioautography results and the UV results indicated that the presence of UV chromophores is not a prerequisite for the detection of antibacterial-like activity. CONCLUSIONS: A reliance on spectrophotometric techniques in the detection of bioactive FQEMs in the environment may underestimate component antibacterial-like activity and, possibly, total antibacterial-like activity expressed by EF and its FQEMs. The described bioautography method provides a screening technique with which antibacterial-like activities derived from EF and possibly other FQs can be detected directly on silica gel HPTLC plates. RECOMMENDATIONS: It is recommended that both bioassay and instrumental analytical techniques be used in any measurement of hazard and risk relating to antibacterial-like activities in the environment that are derived from fluoroquinolone antibiotics and their environmental metabolites.

The response of giant reed (Arundo donax L.) to selenium (Se), added as selenate, was studied. The development, stress response, uptake, translocation, and accumulation of Se were documented in three giant reed ecotypes STM (Hungary), BL (USA), and ESP (Spain), representing different climatic zones. Plantlets regenerated from sterile tissue cultures were grown under greenhouse conditions in sand supplemented with 0, 2.5, 5, and 10 mg Se kg⁻¹ added as sodium selenate. Total Se content was measured in different plant parts using hydride generation atomic fluorescence spectroscopy. All plants developed normally in the 0–5.0 mg Se kg⁻¹ concentration range regardless of ecotype, but no growth occurred at 10.0 mg Se kg⁻¹. There were no signs of chlorosis or necrosis, and the photosynthetic machinery was not affected as evidenced by no marked differences in the structure of thylakoid membranes. There was no change in the maximum quantum yield of photosystem II (Fᵥ/Fₘ ratio) in the three ecotypes under Se stress, except for a significant negative effect in the ESP ecotype in the 5.0 mg Se kg⁻¹ treatment. Glutathione peroxidase (GPx) activity increased as the Se concentration increased in the growth medium. GPx activity was higher in the shoot system than the root system in all Se treatments. All ecotypes showed great capacity of take up, translocate and accumulate selenium in their stem and leaf. Relative Se accumulation is best described as leaf ˃˃ stem ˃ root. The ESP ecotype accumulated 1783 μg g⁻¹ in leaf, followed by BL with 1769 μg g⁻¹, and STM with 1606 μg g⁻¹ in the 5.0 mg Se kg⁻¹ treatment. All ecotypes showed high values of translocation and bioaccumulation factors, particularly the ESP ecotype (10.1 and 689, respectively, at the highest tolerated Se supplementation level). Based on these findings, Arundo donax has been identified as the first monocot hyperaccumulator of selenium, because Se concentration in the leaves of all three ecotypes, and also in the stem of the ESP ecotype, is higher than 0.1% (dry weight basis) under the conditions tested. Tolerance up to 5.0 mg Se kg⁻¹ and the Se hyperaccumulation capacity make giant reed a promising tool for Se phytoremediation.