ZnWO4 nanocrystals: synthesis, morphology, photoluminescence and photocatalytic properties.

The present joint experimental and theoretical work provides in-depth understanding on the morphology and structural, electronic, and optical properties of ZnWO4 nanocrystals. Monoclinic ZnWO4 nanocrystals were prepared at three different temperatures (140, 150, and 160 °C) by a microwave hydrothermal method. Then, the samples were investigated by X-ray diffraction with Rietveld refinement analysis, field-emission scanning electron microscopy, transmission electronic microscopy, micro-Raman and Fourier transform infrared spectroscopy, ultraviolet-visible spectroscopy, and photoluminescence measurements. First-principles theoretical calculations within the framework of density functional theory were employed to provide information at the atomic level. The band structure diagram, density of states, Raman and infrared spectra were calculated to understand the effect of structural order-disorder on the properties of ZnWO4. The effects of the synthesis temperature on the above properties were rationalized. The band structure revealed direct allowed transitions between the VB and CB and the experimental results in the ultraviolet-visible region were consistent with the theoretical results. Moreover, the surface calculations allowed the association of the surface energy stabilization with the temperature used in the synthesis of the ZnWO4 nanocrystals. The photoluminescence properties of the ZnWO4 nanocrystals prepared at 140, 150, and 160 °C were attributed to oxygen vacancies in the [WO6] and [ZnO6] clusters, causing a red shift of the spectra. The ZnWO4 nanocrystals obtained at 160 °C exhibited excellent photodegradation of Rhodamine under ultraviolet light irradiation, which was found to be related to the surface energy and the types of clusters formed on the surface of the catalyst.

Microwave-assisted hydrothermal synthesis of Ag2(W(1-x)Mox)O4 heterostructures: Nucleation of Ag, morphology, and photoluminescence properties.

Ag2W(1-x)MoxO4 (x=0.0 and 0.50) powders were synthesized by the co-precipitation (drop-by-drop) method and processed using a microwave-assisted hydrothermal method. We report the real-time in situ formation and growth of Ag filaments on the Ag2W(1-x)MoxO4 crystals using an accelerated electron beam under high vacuum. Various techniques were used to evaluate the influence of the network-former substitution on the structural and optical properties, including photoluminescence (PL) emission, of these materials. X-ray diffraction results confirmed the phases obtained by the synthesis methods. Raman spectroscopy revealed significant changes in local order-disorder as a function of the network-former substitution. Field-emission scanning electron microscopy was used to determine the shape as well as dimensions of the Ag2W(1-x)MoxO4 heterostructures. The PL spectra showed that the PL-emission intensities of Ag2W(1-x)MoxO4 were greater than those of pure Ag2WO4, probably because of the increase of intermediary energy levels within the band gap of the Ag2W(1-x)MoxO4 heterostructures, as evidenced by the decrease in the band-gap values measured by ultraviolet-visible spectroscopy.

Gene expression and immunolocalization of fibroblast growth factor 2 in the ovary and its effect on the in vitro culture of caprine preantral ovarian follicles.

This study quantified Fibroblast growth factor 2 (FGF-2) mRNA and localized FGF-2 protein in different categories of follicles isolated from goat ovaries. In addition, we verified the effects of this factor on the in vitro culture of preantral follicles isolated from goats. For mRNA quantification, we performed real-time PCR using primordial, primary and secondary follicles, as well as cumulus-oocyte complexes (COCs) and mural granulosa and theca cells of small and large antral follicles. For FGF-2 protein localization, the ovaries were subjected to conventional immunohistochemical procedures. Preantral follicles were isolated and cultured in vitro for 12 days in either control (basic) or supplemented with FGF-2 medium. The expression of FGF-2 mRNA was detected in all categories of follicles and there was no difference in preantral follicles and COCs or granulosa/theca cells from small and large antral follicles. However, in large antral follicles, COCs showed expression levels significantly lower than in granulosa/theca cells (p < 0.05). We observed moderate expression of FGF-2 protein in preantral follicles but not in granulosa cells of primordial follicles and theca cells of secondary follicles. In both small and large antral follicles, strong, moderate and weak staining was observed in oocytes, granulosa and theca cells, respectively. The addition of FGF-2 caused a significant increase in the daily follicular growth rate compared to the control group. We conclude that FGF-2 mRNA is expressed throughout follicular development and that its protein can be found in different patterns in preantral and antral follicles. Furthermore, FGF-2 increases the follicular growth rate in vitro.

Vascular endothelial growth factor-A(165) (VEGF-A(165)) stimulates the in vitro development and oocyte competence of goat preantral follicles.

The aim of this study was to evaluate the effect of vascular endothelial growth factor-A(165) (VEGF-A(165)) on the in vitro development of goat secondary preantral follicles. Preantral follicles (≥150 µm in diameter) were isolated from the ovaries of adult mixed-breed goats and individually cultured for 18 days in αMEM in the absence (control) or presence of VEGF-A(165) at concentrations of 10 ng/ml (VEGF10) and 100 ng/ml (VEGF100). Analyses of follicular survival, diameter, antrum formation and rate of daily growth were performed every 6 days. At the end of the culture period, morphologically normal oocytes (≥110 µm in diameter) were taken for in vitro maturation (IVM). The results demonstrated that all follicles presented oocytes and granulosa cells that were morphologically normal and after labeling with calcein-AM, high rates of oocyte viability were observed in all treatments. The follicular diameter and the growth rate achieved in the presence of VEGF10 were higher than those of the control. Both treatments with VEGF-A(165) showed higher rates of oocyte recovery for IVM when compared with the control. Moreover, only the addition of VEGF-A(165) permitted oocytes grown in vitro to reach metaphase II. Thus, the addition of VEGF-A(165) to the culture medium improves the development of goat preantral follicles cultured in vitro, allowing the production of mature oocytes.

Evaluation of trypsin treatment on the inactivation of bovine herpesvirus type 1 on in vitro produced pre-implantation embryos.

The aim of this study was to evaluate the efficiency of trypsin treatment on the inactivation of bovine herpesvirus type 1 (BoHV-1) on in vitro produced by fertilization and artificially infected bovine embryos. Bovine embryos on day 7 were exposed with 10 microl of BoHV-1, Los Angeles strain 10(7.5) TCID. These embryos and control embryos were divided in two groups: submitted to the sequential washes or to the trypsin treatment according to the International Embryo Transfer Society (IETS) guidelines. The embryos and the last washing drop of each group were used as inoculum to infect Madin Darby bovine kidney (MDBK) cells and submitted to nested PCR reaction using the primer that encodes the gene conserved region of virus glycoprotein gB. The data have shown that the control embryos and their last washing drop were negative. The exposed embryos that were treated with trypsin have shown positive results on the n-PCR and MDBK culture, and their last washing drop were negative. Our data have demonstrated that the trypsin treatment was not able to eliminate the BHV-1 of the embryos, suggesting an interaction between virus and embryo.

Influence of osteopontin in bovine uterine tube fluid on sperm binding and fertilization in RCA-1 lectin-treated oocytes.

This study was conducted to determine the effect of pre-exposure of oocytes to Ricinus communis (RCA-1) lectin and osteopontin (OPN) in uterine tube fluid (UTF) on in vitro sperm-egg binding and fertilization. In vitro-matured bovine oocytes were incubated (39 degrees C, 5% CO(2) in air) for 2 h in the following treatments: (i) 500 microl of fertilization medium (FM); (ii) 250 microl of FM with 0.25 ml of non-luteal ampullary uterine tube fluid (NLAUTF); (iii) 250 microl of FM with 250 microl of NLAUTF and 4 microl of RCA-1 lectin; (iv) 250 microl of FM with 250 microl of NLAUTF, a rabbit polyclonal antibody (1:200) against purified bovine milk OPN, and RCA-1 lectin; (v) 500 microl of FM and RCA-1 lectin. Following incubation, oocytes were washed, placed in FM with 2 microg heparin, and incubated with 1 x 10(5) frozen-thawed spermatozoa per 10 oocytes. Oocytes used to assess sperm binding were stained with Hoescht 33342, and the number of sperm bound per zona pellucida counted. The remaining oocytes were fixed in acid alcohol, stained with 1% acetate-orcein and observed to determine the presence of pronuclei. More sperm bound to the zona pellucida (mean +/- SEM) when oocytes were incubated in treatment 3 (59.0 +/- 5.5) than in treatments 2 (46.4 +/- 5.6), 4 (18.1 +/- 5.4), 5 (33.4 +/- 5.6) or 1 (32.5 +/- 5.6). More oocytes were fertilized when incubated in treatment 3 (91% +/- 3.0) than in 2 (84% +/- 3.0), 4 (40% +/- 3.0), 5 (77% +/- 3.0) or 1 (76% +/- 3.0). As in previous studies, this study suggests that RCA-1 lectin enhances binding of UTF-derived OPN to bovine oocytes, resulting in increased sperm-egg binding and fertilization in vitro and a possible role in fertilization.

Use of polishing pond effluents to cultivate lettuce (Lactuca sativa) in a hydroponic system.

The sanitary quality and productivity of hydroponic lettuce (Lactuca sativa L.) plants cultivated under greenhouse conditions and treated with effluent from anaerobic reactor + polishing pond followed by physical-chemical treatment was evaluated. Two hydroponic cultivations were performed at summer and winter time at Vitoria-ES, Brazil. The treatments for both cultivations were: T1) conventional nutrient solution, T2) effluent from physical-chemical treatment, T3) effluent from polishing pond, and T4) effluent from polishing pond with 50% dilution. The plants were evaluated for microbial contamination, productivity and nutrient content. In all cases, no significant microbial contamination of lettuce was detected and the levels of macronutrients in the shoot system were similar to those in published reports. In the experiments from summer season, the treatments T1 and T2 resulted in higher production than the T3 and T4 treatments. Plants from T3 and T4 had a less developed root system as a result of reduced oxygenation from competition with the higher algae biomass content from the polishing pond effluent. In the winter season, the effect of the algal biomass was pronounced only in the T3 treatment (undiluted effluent from polishing pond). In conclusion, hydroponic cultivation of lettuce with pond effluent is suitable as a complement to water and nutrients for plants.

Oviductal fluid proteins associated with the bovine zona pellucida and the effect on in vitro sperm-egg binding, fertilization and embryo development.

Studies have demonstrated that oviductal fluid (ODF) proteins associate with eggs of numerous species including the bovine. In this study, the association of three ODF proteins, the bovine oestrus-associated protein, osteopontin (OPN), lipocalin-type prostaglandin D synthase (L-PGDS), with the bovine zona pellucida (ZP) was demonstrated by immunohistochemistry and western blot. The biological function of ODF derived egg-associated OPN and L-PGDS in sperm binding, fertilization and embryonic development was also explored. In vitro matured bovine oocytes were pre-incubated with ODF collected by cannula from cows in oestrus, or ODF with antibodies to OPN, L-PGDS and bovine serum albumin (BSA). Following incubation, oocytes were inseminated with 1 x 10(5) frozen-thawed spermatozoa, and they were evaluated for sperm binding, fertilization and embryonic development in vitro. Pre-treatment of ODF with antibodies to all of proteins reduced sperm binding to the ZP and fertilization in vitro. Cleavage rates were not significantly different among incubations, but rates of embryo development were significantly decreased. We conclude that antibodies to OPN, L-PGDS and BSA react with oocytes incubated with ODF and inhibit sperm binding, fertilization and embryonic development in vitro, suggesting a potential role of these proteins in these events.

Pre-treatment of cattle sperm and/or oocyte with antibody to lipocalin type prostaglandin D synthase inhibits in vitro fertilization and increases sperm-oocyte binding.

The present study was conducted to determine the affect of pre-treating of oocytes and/or sperm with a rabbit polyclonal antibody against recombinant cattle lipocalin type prostaglandin D synthase (alpha L-PGDS) on in vitro sperm-oocyte binding and fertilization. In vitro matured cattle oocytes were incubated (39 degrees C, 5% CO(2) in air) for 1h in the following treatments either 500 microL of fertilization medium (FM) or FM with alpha L-PGDS (1:2000). Frozen-thawed spermatozoa were washed by a 45/90% layered Percoll gradient centrifugation and incubated for 1h either FM or FM with alpha L-PGDS. This study utilized five different treatments: (1) no antibody (control); (2) a rabbit IgG against a non-bovine antigen, bacterial histidase (alpha-hist); (3) alpha L-PGDS at fertilization time (with fertilization medium); (4) alpha L-PGDS-treated oocytes; or (5) alpha L-PGDS-treated sperm. Pre-treated oocytes were incubated with 10 x 10(4) washed spermatozoa per 25 oocytes. Oocytes used to assess sperm binding were stained with Hoescht 33342, and the number of sperm bound per zonae pellucidae counted. The remaining oocytes were fixed in acid alcohol, stained with 1% acetate-orcein and observed to determine the presence of pronuclei. More sperm bound to the zonae pellucidae when oocytes and/or sperm were pre-treated with alpha L-PGDS: (1) 26.4+/-3.0; (2) 25.6+/-3.0; (3) 59.7+/-3.0; (4) 56.4+/-3.0; and (5) 57.1+/-3.0. Addition of alpha L-PGDS with sperm, oocytes, or both, decreased fertilization (P<0.05) compared with the control: (1) 89.2+/-2.0%; (2) 87.5+/-2.0%; (3) 19.4+/-2.0%; (4) 27.2+/-3.1%; and (5) 14.1+/-3.4%. The alpha L-PGDS reacts with both oocytes and spermatozoa, resulting in increases of in vitro sperm-oocyte binding and inhibition of fertilization. These observations suggest that L-PGDS may have a role in cattle fertilization.

Pre-treatment of cattle semen or oocytes with purified milk osteopontin affects in vitro fertilization and embryo development.

This study was designed to investigate the effects of pre-incubating cattle spermatozoa or matured oocytes with purified osteopontin (OPN) from cattle milk on fertilization in cattle and embryonic development in vitro. There were two different experiments, semen from six mature Holstein bulls (Bos Taurus) was frozen with different concentrations of OPN (0, 1, 10, 100 microg/mL). Matured cattle oocytes were also pre-treated with OPN (0, 10, 100 microg/mL). In both experiments, pre-treated oocytes or frozen semen, was processed for in vitro fertilization and embryo development. Significantly more oocytes were fertilized when using frozen semen with 10 microg/mL OPN (bull 2=85+/-4% and bull 5=78+/-4%) than without OPN (bull 2=75+/-4% and bull 5=69+/-4%). Those bulls also had increase in cleavage and embryo development (bull 2=85+/-3%, 41+/-1.9%; bull 5=76+/-2%, 37+/-1.8%) compared with control (bull 2=75+/-3%, 30+/-2%; bull 5=68+/-2%, 29+/-2%). Incubating matured oocytes in 10 microg/mL OPN (87+/-3%) and 100 microg/mL OPN (88+/-3%) significantly increased fertilization than control (73+/-3%). OPN also improve cleavage, and embryo development in treatments with 10 microg/mL OPN (82.7+/-1.3%; 31.7+/-1.4%) and 100 microg/mL OPN (85.8+/-1.3%; 33.8+/-1.5%) when compared with control (74.1+/-1.3%; 24.2+/-1.2%). These data suggest that both, spermatozoa from some bulls and oocytes may associate with OPN, suggesting a facilitory role on in vitro fertilization and embryo development.

Influence of arginine-glycine-aspartic acid (RGD), integrins (alphaV and alpha5) and osteopontin on bovine sperm-egg binding, and fertilization in vitro.

Osteopontin (OPN), a phosphoprotein containing an arginine-glycine-aspartic acid (RGD) sequence, has been identified in cow oviduct epithelium and fluid. To investigate the potential role OPN in fertilization, we evaluated the ability of RGD peptide (arginine-glycine-aspartic), RGE peptide (arginine-glycine-glutamic acid), integrins alphaV and alpha5 antibodies and OPN antibody to influence bovine in vitro sperm-egg binding and fertilization. Treatment of sperm or oocytes with the RGD peptide prior fertilization significantly decreased in vitro sperm-egg binding and fertilization compared to the non-treated controls or those treated with RGE peptide. Binding and fertilization were also significantly decreased when in vitro matured bovine oocytes or sperm were pre-incubated with integrins alphaV and alpha5 antibodies at concentration ranging from 5 to 20 microg/mL. Addition of a rabbit polyclonal IgG antibody against purified bovine milk OPN with sperm or/and oocytes decreased (P<0.05) fertilization compared to the in vitro-fertilized control. These data provided evidence that integrin ligands existed on bovine oocytes and spermatozoa that contained RGD recognition sequences, and that antibody to OPN, a protein that contains that RGD sequence, was capable of reducing sperm-egg binding and fertilization in vitro.

Modelling hydrogen sulphide emission in a WWTP with UASB reactor followed by aerobic biofilters.

Hydrogen sulphide (H2S) represents one of the main odorant gases emitted from wastewater treatment plants (WWTP) and a mathematical model can be a fast and low cost tool to estimate its emission. In this work H2S emission rates in a WWTP, composed of an up-flow anaerobic sludge blanket (UASB) reactor and an aerobic biofilter (BF), are estimated using four mathematical models available in the literature (AP-42, GPC, TOXCHEM + and WATER8). The results show that the GPC model leads to the best agreement with the experimental data, except for the biofilter due to its lack of capability to include biodegradation as a H2S removal process. On the other hand, the AP-42 and WATER8 models showed a slightly better ability to predict H2S removal in the biofilter than the TOXCHEM + model, as all models underestimate the H2S concentration decay.

Alkaline and acid hydrolytic processes in aerobic and anaerobic sludges: effect on total EPS and fractions.

Sludge samples from an upflow anaerobic sludge blanket (UASB) reactor and four submerged aerated biofilters (BFs) of a wastewater treatment plant (1,000 inhab.) were processed at bench scale by alkaline and acid hydrolysis with the objective to evaluate the organic matter solubilization, volatile solids (VS) destruction and the effect of hydrolytic processes on the extracellular polymeric substances (EPS) fraction of the sludge samples. The results showed that alkaline hydrolysis of sludge samples treatment with 1.0% total solids (TS) using NaOH 20 meq L(-1) was more efficient on organic matter solubilization and VS destruction than acid hydrolysis. The EPS sludge content was also affected by the alkaline treatment of anaerobic sludge samples. The EPS concentrations (mg EPS/gVSS) on the anaerobic sludge after the alkaline treatment were significantly lowered according to sample height in the UASB reactor. Data indicated that the EPS sludge fraction is the main component affected by the alkaline hydrolytic process of anaerobic sludge samples.

Pathogen removal efficiency from UASB + BF effluent using conventional and UV post-treatment systems.

The aim of this study was to verify the efficiency of removal of microorganisms in effluents of a Wastewater Treatment Plant (WWTP) comprising an association of a UASB reactor followed by three submerged aerated biofilters (BAF) and one tertiary filter. The WWTP designed to treat domestic wastewater from a population of 1,000 inhabitants showed high removal efficiency for organic matter and suspended solids. Helminth eggs were also efficiently removed from the tertiary effluent and were found in the sludge from the UASB reactor; however, removal of bacteria in this system was very low. To enhance the efficiency of the system, the effluent from tertiary filters was submitted to UV disinfection in a real scale reactor. Our results showed that UV irradiation was very effective at lowering the concentrations of E. coli, thermotolerant coliforms and coliphages to acceptable levels for agricultural reuse. Salmonella spp. and helminth eggs were seeded into the tertiary effluent before passing through the UV reactor. Salmonella was not found in the final effluent, but helminth eggs were not completely inactivated by UV irradiation and viable eggs were detected after 28 d of incubation.

Inactivation of Salmonella spp. from secondary and tertiary effluents by UV irradiation.

The aim of this study was to verify the efficiency of UV irradiation in the inactivation of Salmonella spp. in treated wastewater with different levels of turbidity and exposed to increasing doses of UV irradiation. Experiments were carried out in a batch reactor and in a real scale reactor. Salmonellae obtained from clinical samples were seeded into autoclaved wastewater collected from a wastewater treatment plant (WWTP) comprising an association of a UASB reactor followed by three submerged aerated biofilters (BAF) and one tertiary filter. The results showed that salmonellae were not inactivated in effluents from the UASB reactor indicating that the presence of suspended solids was an important obstacle to UV penetration in bacteria. However, UV irradiation was efficient in inactivating Salmonella of effluents from aerated secondary and tertiary biofilm reactors.