Extraction of seed oil from Diospyros lotus optimized using response surface methodology

Oil from seeds of Diospyros lotus was extracted using a conventional method with two different solvents:hexane and petroleum ether. A central composite design with response surface methodology were used to optimize the process. A second-order polynomial equation was employed, and ANOVA was applied to evaluate the impact of various operating parameters including extraction temperature(x_1; 44.9–70.1 °C), extraction time(x_2;5.0–10.0 h) and solvent to solid ratio(x_3;11.6–28.4 mL g^-1), on oil yield. Experiments to validate the model showed decent conformity between predicted and actual values. Extraction conditions for optimal oil yield were 61 °C, 8.75 h extraction duration and 19.25 mL g^-1 solvent to solid ratio. Under these conditions, the oil yield was predicted to be 5.1340%. Oil samples obtained were then analyzed using gas chromatography. The fatty acid composition revealed the major fatty acids to be oleic acid(C18:1) and linoleic acid(C18:2). The analysis of oil also demonstrated a decent ratio between omega-3 and omega-6 fatty acids. The structure of seeds was imaged using scanning electron microscopy. Oil quality was analyzed thermogravimetrically and by Fourier transform infrared spectroscopy. The assigned nutritional features of the D. lotus oil suggested that it can be used as an edible oil in pharmaceutical and food industry in the future.     

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Background: Pseudomonas aeruginosa is one of the opportunistic pathogens causing frequent hospital-acquired life-threatening infections in mechanically ventilated patients. The most significant virulence factor of P. aeruginosa is T3SS. PcrV is an important structural protein of the T3SS. Methods:In the current investigation, a recombinant scFv mAb against the PcrV protein was expressed in EnBase® (fed-batch) cultivation mode. The pETiteTM N-His SUMO Kan vector, including anti-PcrV scFv gene, was transformed into Escherichia coli (BL21) cells. The expression and solubility of anti-PcrV scFv protein were investigated at two different temperatures (25 °C and 30 °C) and at different induction times (4, 6, 8, 12, and 24 hours). Results:Increased efficiency was achieved by EnBase® compared to LB broth; owing to the slow release of glucose, the maximum level of solubility and total protein expression was observed in EnBase® cultivation system at 30 °C and 24 h post induction. Furthermore, IC₅₀ for anti-PcrV scFv protein was determined to be approximately 7 μg/mL. Conclusion:Anti-PcrV scFv produced in this study showed promising in vitro results, protecting RBC from lysis by P. aeruginosa (exoU+). Key Words: Fed Batch, recombinant protein, Pseudomonas aeruginosa, scFv     

The effect of pH and calcium on copper availability to the springtail Folsomia candida in simplified soil solutions

The effect of pH and calcium on copper bioavailability to the springtail Folsomia candida was determined by assessing uptake kinetics upon copper exposure for 15 days in simplified soil solutions. A slight bioaccumulation of copper was observed in all treatments and controls. The effect of exposure concentration and calcium level on copper accumulation was not significant. Although pH and time slightly but significantly affected copper concentrations in the body, copper uptake rates were not significantly different from zero. This suggests that copper uptake in F. candida exposed to sub-lethal copper concentrations in soil pore water is not affected by pH and calcium.     

Study of interleukin-10 and interleukin-12 productions in response to lipopolysaccharides extracted from two different Brucella strains

The aim of the present study is to investigate the cytokines induction by smooth lipopolysaccharides (S-LPS) extracted from Brucella melitensis (Rev1 vaccine strain) and Brucella abortus (a field isolate). These lipopolysaccharides were used to induce inflammatory cytokines production in peripheral blood cell culture of healthy individuals. Secretion of IL-10 and IL-12 (p70) were measured by means of specific Elisa. In addition, intracellular expression of IL-12 was assessed in CD14+ cells by flow cytometry. It was shown that Brucella LPS is a potent inducer of IL-10. However interferon gamma (IFN-gamma) priming was able to significantly decrease the production of IL-10. Flow cytometry studies showed that LPS alone was not able to induce intracellular IL-12 expression in CD14+ cells. Nevertheless, IFN-gamma priming significantly increased the percentage of CD14+ IL-12+ cells. In conclusion, it was demonstrated that the Brucella LPS could be a potent inducer of IL-10 and induction of IL-12 production needs the most favorable conditions.     

Chemical confirmatory tests for ochratoxin A, citrinin, penicillic acid, sterigmatocystin, and zearalenone performed directly on thin layer chromatographic plates

Published tests have been improved and a new procedure is described for chemical confirmation of mycotoxins directly on thin layer plates. After extraction and preliminary cleanup chromatography with n-hexane or chloroform, the mycotoxins ochratoxin A, citrinin, penicillic acid, sterigmatocystin, and zearalenone were easily separated by thin layer chromatography (TLC) using toluene-ethyl acetate-90% formic acid (6 + 3 + 1) developing solvent. In chemical confirmatory methods, the developed chromatogram was exposed to vapors of pyridine, acetic anhydride, or a mixture, or the mycotoxins were over-spotted. With this treatment, ochratoxin A, citrinin, penicillic acid, and zearalenone were converted to new fluorescent compounds, and observed under 365 nm light after re-chromatography with the same developing solvent. Sterigmatocystin was confirmed chemically using TLC plates impregnated with 0.6N H2SO4 or 10% oxalic acid in methanol. The described procedures are satisfactory for confirming mycotoxins present in standards, artificially contaminated grain samples (barley, corn, oat, rye, and wheat), and extracts from both fungal cultures and naturally contaminated grain samples.     

Genetic diversity of two Philippine native freshwater goby species (Perciformes: Gobiidae): implications for conservation

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Effect of pistachio (Pistacia vera) hull extract on growth performance,body composition,total phenolic compound and fillets peroxide value of common carp,Cyprinus carpio

This study was carried out to investigate the effect of pistachio green hull extract (PGHE) on growth, survival rates, body composition and oxidative spoilage of common carp fillet. Three hundred fish (11.65 ± 1.65 g) were divided into five dietary treatments with three replications, including 0, 0.5, 1.5, 4.5 and 9 g PGHE kg^?1 of diet. No significant difference were observed in specific growth rate, food conversion rate, weight gain percentage, survival rate and body composition (i.e. moisture, protein, lipid and ash) between different treatments. By increasing the level of PGHE, amount of phenolic compounds in fish meat was elevated whilst peroxide value was reduced. The amount of phenolic compounds in fish fillet was significantly lower (P < 0.05) in control treatment in comparison with other treatments. Peroxide content for 9 g PGHE kg^?1 diet treatment (6.15 ± 0.79 meq peroxide kg^?1 oil) was significantly (P < 0.05) lower than control group (18.82 ± 1.93 meq peroxide kg^?1 oil). Regression analysis of peroxide value indicated that minimum peroxide value obtains in 1187 mg gallic acid kg^?1 diet. This study indicated that ethanolic extract of Pistachia vera hull can be considered as a supplement in fish diet to delay oxidative spoilage of common carp fillet.     

Head blight and biosynthesis of Fusarium toxins in barley kernels field inoculated withFusarium culmorum

Heads of 12 spring barley genotypes (eight cultivars, four lines) were inoculated with conidial suspensions of twoFusarium culmorum isolates (I₁ and I₂). Both isolates caused the following significant reductions (%) when compared with the control: number of kernels head^–1 12 and 16, weight of 1000 kernels 47 and 24, yield 49 and 37, for I₁ and I₂ respectively. Both isolates were able to produce average levels (mg kg^–1) of deoxynivalenol of 67.1 and 13.9, 3-acetyldeoxynivalenol of 9.4 and 1.4 and zearalenone of 0.3 and 0.4 for I₁ and I₂ respectively. In kernels of three genotypes inoculated with the same isolates, 15-acetyldeoxynivalenol at an average level (mg kg^–1) of 1.3 and 1.5 was detected, while in kernels of six genotypes inoculated with I₂, 0.2 of nivalenol was found. A significant correlation for yield and toxin level was found for less pathogenicF. culmorum isolate I₂.     

The effect of multiplex-PCR-assessed major pathogens causing subclinical mastitis on somatic cell profiles

The major pathogens causing mastitis were evaluated by multiplex-polymerase chain reaction (M-PCR) with self-designed primers in four quarters of the first, third, and fifth parities in industrial, semi-industrial, and traditional dairy cattle farms in Iran. With the incidence of infection in the quarters by Staphylococcus aureus and Streptococcus agalactiae, the mean log somatic cell count (log SCC) increased from 5.06 to 5.77. The smallest changes occurred with Escherichia coli. Contagious pathogens, when compared with environmental pathogens, were more prevalent and common and created more profound quantitative and qualitative changes in SCC profiles. The second part of the study surveyed the diversity of contaminating pathogens and their effect on quantitative and qualitative profiles of somatic cells. M-PCR was used to determine the absence (M-PCR(-)) and presence of one (M-PCR(+1)), two (M-PCR(+2)), and three (M-PCR(+3)) major pathogens in raw milk samples. Quarter log SCC increased from 5.06 (for M-PCR(-1)) to 5.5 (for M-PCR(+1)), 5.7 (for M-PCR(+2)), and 6 (for M-PCR(+3)). Percent changes in polymorphonuclears (PMNs) were not significant between different quarters and parities but were significant between different farms in terms of pathogen diversity (P?相似文献   

Essential oil variation among and within six Achillea species transferred from different ecological regions in Iran to the field conditions

The compositions of essential oils of 19 accessions belonging to six different Achillea species, transferred from the natural habitats in 10 provinces of Iran to the field conditions, were assessed. The relationship between the leaf areas of selected accessions with their essential oil content was also investigated. Essential oil yield of dried plants obtained by hydro-distillation ranged from 0.1 to 2.7% in leaves. Results indicated a significant variation in oil composition among and within species. Total of 94 compounds were identified in 19 accessions belonging to the six species of A. millefolium, A. filipendulina, A. tenuifolia, A. santolina, A. biebersteinii and A. eriophora. The major constituents of the leaves in the tested genotypes were determined as germacrene-D, bicyclogermacrene, camphor, borneol, 1,8-cineole, spathulenol and bornyl acetate. According to the major compounds, four chemotypes were defined as: (I) spathulenol (1.64–34.31%) + camphor (0.2–15.61%) (7 accessions); (II₁) germacrene-D (18.78–23.93%) + borneol (7.93–8.26%) + bornyl acetate (11.56–14.66%) (5 accessions); (II₂) germacrene-D (13.28–36.28%) + bicyclogermacrene (5.93–8.4%) + 1,8-cineole (15.26–19.41%) + camphor (14.95–23.32%) (2 accessions); (III) borneol + camphor (52.04–63.27) (2 accessions); (IV) germacrene-D (45.86–69.64%) (3 accessions). The relationships of chemotypes with soil type and climatic conditions of collected regions were assessed, as probable reasons of high variations in essential oil components, and discussed.