Isolation and molecular characterization of Cryptococcus species isolated from pigeon nests and Eucalyptus trees.

BACKGROUND AND PURPOSE: Cryptococcus species are pathogenic and non-pathogenic basidiomycete yeasts that are found widely in the environment. Based on phenotypic methods, this genus has many species; however, its taxonomy is presently being re-evaluated by modern techniques. The Cryptococcus species complex includes two sibling taxa of Cryptococcus neoformans and Cryptococcus gattii. We aimed to investigate the possible distribution of Cryptococcus species in pigeon nests and Eucalyptus trees in Ilam, Iran, using molecular techniques. MATERIALS AND METHODS: Two hundred and seventy-four specimens were collected from pigeon nests and Eucalyptus trees during 2016-2017. All the specimens were sub-cultured on Sabouraud Glucose Agar with chloramphenicol and bird seed agar. For molecular identification, the ITS15.8SITS2 rDNA region was amplified using the first and fourth internal transcribed spacer (ITS1 and ITS4, respectively) primers. The purified products were applied for cycle sequencing reactions in forward direction with ITS1 primer. The obtained results were analyzed with Chromas 2.3. RESULTS: Thirty-three out of 186 cultures (17.7%) and 11 out of 88 cultures (12.5%) were positive among pigeon nest and Eucalyptus tree specimens, respectively. Cryptococcus albidus (17.2%), C. albidus var. kuetzingii (3.4%), C. adeliensis (3.4%), C. uzbekistanensis (3.4%), and C. neoformans var. grubii (3.4%) were isolated from pigeon nests, and Cryptococcusadeliensis (25%) was the only Cryptococcus species isolated from Eucalyptus trees. CONCLUSION: The presence of pigeons and Eucalyptus trees in the vicinity of some particular places such as rest homes and hospitals should be considered as a risk factor for the immunocompromised population.

N,N-dimethylhexadecyl carboxymethyl chitosan as a potential carrier agent for rotenone.

In this study, an amphiphilic chitosan derivative namely N,N-dimethylhexadecyl carboxymethyl chitosan (DCMC) was synthesised and applied for the first time as a carrier agent for rotenone. The physical and chemical properties of DCMC were characterised by using Fourier Transform Infrared Spectrometer (FTIR), Proton Nuclear Magnetic Resonance Spectrometer ((1)H NMR), CHN-O Elemental Analyser, Thermogravimetric Analyser (TGA) and Differential Scanning Calorimeter (DSC). DCMC was soluble in acidic (except pH 4), neutral and basic media with percent of transmittance (%T) values ranged from 67.2 to 99.4%. The critical micelle concentration (CMC) was determined as 0.095mg/mL. Transmission Electron Microscopy (TEM) analysis confirmed that DCMC has formed self-aggregates and exhibited spherical shape with the size of 65.5-137.0nm. The encapsulation efficiency (EE) and loading capacity (LC) of DCMC micelles with different weight ratios (DCMC:rotenone; 5:1, 50:1 and 100:1) were determined by using High Performance Liquid Chromatography (HPLC). The weight ratio of 100:1 gave the best EE with the value of more than 95.0%. DCMC micelles performed an excellent ability to control the release of rotenone, of which 99.0% of rotenone was released within 48h. Overall, DCMC has several key features to be an effective carrier agent for pesticide formulations.

Thyroid endocrine status of wild European eels (Anguilla anguilla) in the Loire (France). Relationships with organic contaminant body burdens.

In teleost fish, thyroid function is involved in various critical physiological processes. Given the complexity of the hypothalamo-pituitary-thyroid (HPT) axis, a large number of genes and proteins can be the potential target of endocrine-disrupting compounds (EDCs). The aim of this study was to evaluate, in yellow and silver European eels (Anguilla anguilla), potential effects of EDCs on thyroid status by analyzing the associations between EDC body burdens and thyroid hormones (THs). In yellow individuals, greater free T3/T4 ratios (FT3/FT4) and lower plasma FT4 levels were associated with greater concentrations of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs), as highlighted by significant correlations with many congeners. Few positive relationships with alkylphenols were noticed. In contrast, silver eels usually exhibited less significant correlations between THs and contaminant loads. Expression of a series of genes involved in the HPT axis was also investigated in the silver individuals. Concerning mRNA expression in silver females, some main correlations were noticed: thyroid-stimulating hormone (TSHß) gene expression was significantly correlated to numerous PCBs, and hepatic mRNA levels of deiodinase 2 (Deio 2) were negatively correlated to 2-hydroxyfluorene (2-OHF) and 2-naphtol (2-OHNa). Thyroid receptor (TRα and TRß) mRNA levels exhibited weak negative correlations with some PBDEs in silver females and males. Hepatic vitellogenin (Vtg) mRNA levels were detected in all silver males but at lower levels than in silver females. In males, Vtg mRNA levels were positively associated to FT4/TT4. In silver females, strong positive correlations were found between congeners of PCBs, PBDEs and PFAS suggesting potential estrogenic effects. Overall, the observed results indicate that several organic contaminants, mainly dl-, ndl-PCBs and PBDEs, could be associated with changes in thyroid homeostasis in these fish, via direct or indirect interactions with peripheral deiodination, metabolism of T4 and mechanisms involved in TSHß, Deio 2 and Vtg gene transcription.

Immobilisation of Cu, Pb and Zn in Scrap Metal Yard Soil Using Selected Waste Materials.

Immobilisation of heavy metals in a 30-year old active scrap metal yard soil using three waste materials, namely coconut tree sawdust (CTS), sugarcane bagasse (SB) and eggshell (ES) was investigated. The contaminated soil was amended with amendments at application rates of 0 %, 1 % and 3 % (w/w). The effects of amendments on metal accumulation in water spinach (Ipomoea aquatica) and soil metal bioavailability were studied in a pot experiment. All amendments increased biomass yield and reduced metal accumulation in the plant shoots. The bioconcentration factor and translocation factor values of the metals were in the order of Zn > Cu > Pb. The addition of ES, an alternative source of calcium carbonate (CaCO3), has significantly increased soil pH and resulted in marked reduction in soil metal bioavailability. Therefore, CTS, SB and ES are promising low-cost immobilising agents to restore metal contaminated land.

Occurrence of POPs and other persistent organic contaminants in the European eel (Anguilla anguilla) from the Loire estuary, France.

The chemical contamination of the Loire estuary by three classes of persistent organic pollutants (POPs): the polychlorinated biphenyls (PCBs), the polybrominated diphenyl ethers (PBDEs) and the perfluorinated and polyfluorinated alkyl substances (PFAS), and three families of organic contaminants, the alkylphenols (APs), the polycyclic aromatic hydrocarbon metabolites (OH-PAHs) and the bisphenol A (BPA) were investigated in the muscles and bile of European eel (Anguilla anguilla). Yellow eels (n=30) were caught in three different points along the estuary to highlight variations between sites and sources of contaminations. Silver eels (n=15) were also studied to compare contaminant impregnation between different life stages of the species. Average concentrations in the muscles of the eel ranged between: 857 and 4358 ng/gLW for the PCBs, 26 and 46 ng/gLW for the PBDEs, 130 and 1293 ng/gLW for the PFAS; and in bile: 31 and 286 µg/g protein for the APs, 9 and 26 µg/g protein for the OH-PAHs and ND-1213 µg/g protein for the BPA. Among PCBs, PCB 153 (40% contribution to the sum of PCBs) was predominant in all eel muscles. PBDE 47 (60%) was the most predominant PBDE congeners, while perfluorooctanesulfonic acid (85%) was the most widely detected PFAS. For APs, 4p-nonylphenol (91%) was the most abundant and for the OH-PAHs, it was 1OH-Pyrene (63%). All the eels exceeded the environmental quality standards (EQS) for biota for the PBDEs and about 75% were higher than the EQS specific to PFOS. Finally, 20% of the analyzed eels presented TEQ concentrations above the maximum limits for lipid-rich species. These results supplied new data on the occurrence, levels, and patterns of 53 organic chemicals in the eels from the Loire estuary and they highlighted the need of further investigations focused notably on the potential effects of these chemicals on this species and their analysis in the water and sediments of the estuary.

Biodegradation of chitosan and its effect on metal bioavailability.

The microbial breakdown of chitosan, a fishery waste-based material, and its derivative cross-linked chitosans, in both non-contaminated and contaminated conditions was investigated in a laboratory incubation study. Biodegradation of chitosan and cross-linked chitosans was affected by the presence of heavy metals. Zn was more pronounced in inhibiting microbial activity than Cu and Pb. It was estimated that a longer period is required to complete the breakdown of the cross-linked chitosans (up to approximately 100 years) than unmodified chitosan (up to approximately 10 years). The influence of biodegradation on the bioavailable fraction of heavy metals was studied concurrently with the biodegradation trial. It was found that the binding behaviour of chitosan for heavy metals was not affected by the biodegradation process.

Metal accumulation in Lolium perenne and Brassica napus as affected by application of chitosans.

The effects of chitosan, a fishery waste-based material, and its derivative glutaraldehyde cross-linked chitosan (chitosan-GLA) on metal uptake by Lolium perenne (perennial ryegrass) and Brassica napus (rapeseed) were studied in a greenhouse pot experiment. Metal uptake by perennial ryegrass was highly dependent on the rate of addition of the chitosans. Low application rate (1% w/w) enhanced metal uptake, whereas 10% (w/w) addition decreased metal uptake. It was estimated that chitosan 1% (w/w) treatment could assist perennial ryegrass to remove approximately 3.2 kg Zn/ha and 0.29 kg Pb/ha. For rapeseed, metal uptake was decreased at all rates of application of chitosans. The ammonium acetate extractable metals in soil decreased following application of chitosans and plant growth.

Binding of heavy metal contaminants onto chitosans--an evaluation for remediation of metal contaminated soil and water.

The binding efficiency of chitosan samples for Ag(+), Cd(2+), Cu(2+), Pb(2+) and Zn(2+) has been evaluated in order to consider their application to remediate metal contaminated soil and water. The sorption behaviour of metal ions was assessed using a batch technique at different contact time and initial metal concentration with different background electrolytes. The kinetics followed a pseudo-second-order model, while the equilibrium data correlated well with the Freundlich and Langmuir isotherm models. For example, the maximum sorption capacity (Q) for chitosan was estimated as 1.93 mmol/g for Ag(+), 1.61 mmol/g for Cu(2+), 0.94 mmol/g for Zn(2+), 0.72 mmol/g for Cd(2+) and 0.64 mmol/g for Pb(2+). Covalent interaction between metal ions and functional groups (amino and hydroxyl) of the chitosans was the main binding mechanism. Ion exchange is not an important process. Chitosan and cross-linked chitosans were able to bind metal ions in the presence of K(+), Cl(-) and NO(3)(-). The nature of Cl(-) and NO(3)(-) ions did not affect Zn(2+) binding by the chitosans. Even at 11x dilution, the chitosans were able to retain metal ions on their surfaces.

Chitosan as a potential amendment to remediate metal contaminated soil - a characterisation study.

The potential of chitosan, a fishery waste-based material, as a soil amendment to clean-up metal contaminated soil was investigated. Chitosan was treated using glutaraldehyde (GLA), epichlorohydrin (ECH) and ethylene glycol diglycidyl ether (EGDE) as cross-linking reagents to enhance its chemical stability in acidic media and to improve its physical properties. Cross-linking treatment had significant effects on chitosan surface area, pore diameter, surface morphology and crystallinity. Interaction with Ag(I), Pb(II) and Cu(II) decreased the crystallinity of the materials and changed their surface morphology significantly. FTIR analysis confirmed that N and O atoms served as binding sites for the metal ions. Chitosan and treated chitosans were able to bind metal ions, even in the presence of K(+), Cl(-) and NO(3)(-), which are dominant ions in soil. Therefore, remediation of metal contaminated soil using chitosan and cross-linked treated chitosans as soil amendments is feasible.

Isotherm, kinetic and thermodynamic studies of lead and copper uptake by H2SO4 modified chitosan.

The kinetic and thermodynamic adsorption and adsorption isotherms of Pb(II) and Cu(II) ions onto H(2)SO(4) modified chitosan were studied in a batch adsorption system. The experimental results were fitted using Freundlich, Langmuir and Dubinin-Radushkevich isotherms; the Langmuir isotherm showed the best conformity to the equilibrium data. The pseudo-first order, pseudo-second order and intraparticle diffusion kinetic models were employed to analyze the kinetic data. The adsorption behavior of Pb(II) and Cu(II) was best described by the pseudo-second order model. Thermodynamic parameters such as free energy change (DeltaG degrees ), enthalpy change (DeltaH degrees ) and entropy change (DeltaS degrees ) were determined; the adsorption process was found to be both spontaneous and exothermic. No physical damage to the adsorbents was observed after three cycles of adsorption/desorption using EDTA and HCl as eluents. The mechanistic pathway of the Pb(II) and Cu(II) uptake was examined by means of Fourier transform infrared (FTIR) and Energy dispersive X-ray (EDX) spectroscopy. The equilibrium parameter (R(L)) indicated that chitosan-H(2)SO(4) was favorable for Pb(II) and Cu(II) adsorption.

Adsorption behaviour of Fe(II) and Fe(III) ions in aqueous solution on chitosan and cross-linked chitosan beads.

A batch adsorption system was applied to study the adsorption of Fe(II) and Fe(III) ions from aqueous solution by chitosan and cross-linked chitosan beads. The adsorption capacities and rates of Fe(II) and Fe(III) ions onto chitosan and cross-linked chitosan beads were evaluated. Chitosan beads were cross-linked with glutaraldehyde (GLA), epichlorohydrin (ECH) and ethylene glycol diglycidyl ether (EGDE) in order to enhance the chemical resistance and mechanical strength of chitosan beads. Experiments were carried out as function of pH, agitation period, agitation rate and concentration of Fe(II) and Fe(III) ions. Langmuir and Freundlich adsorption models were applied to describe the isotherms and isotherm constants. Equilibrium data agreed very well with the Langmuir model. The kinetic experimental data correlated well with the second-order kinetic model, indicating that the chemical sorption was the rate-limiting step. Results also showed that chitosan and cross-linked chitosan beads were favourable adsorbers.

Equilibrium and kinetics studies of adsorption of copper (II) on chitosan and chitosan/PVA beads.

The adsorption of Cu(II) ions from aqueous solution by chitosan and chitosan/PVA beads was studied in a batch adsorption system. Chitosan solution was blended with poly(vinyl alcohol) (PVA) in order to obtain sorbents that are insoluble in aqueous acidic and basic solution. The adsorption capacities and rates of Cu(II) ions onto chitosan and chitosan/PVA beads were evaluated. The Langmuir, Freundlich and BET adsorption models were applied to describe the isotherms and isotherm constants. Adsorption isothermal data could be well interpreted by the Langmuir model. The kinetic experimental data properly correlated with the second-order kinetic model, which indicates that the chemical sorption is the rate-limiting step. The Cu(II) ions can be removed from the chitosan and chitosan/PVA beads rapidly by treatment with an aqueous EDTA solution. Results also showed that chitosan and chitosan/PVA beads are favourable adsorbers.