Evidence for a radial strain gradient in apple fruit cuticles

Main conclusion

The morphological outer side of the apple fruit cuticle is markedly more strained than the inner side. This strain is released upon wax extraction. This paper investigates the effect of ablating outer and inner surfaces of isolated cuticular membranes (CM) of mature apple (Malus × domestica) fruit using cold atmospheric pressure plasma (CAPP) on the release of strain after extraction of waxes. Strain release was quantified as the decrease in area of CM discs following CAPP treatment and subsequent solvent extraction of wax. Increasing duration of CAPP treatment proportionally decreased CM mass per unit area. There was no difference in mass loss rate between CAPP treatments of outer or inner surfaces. Also, there was no difference in surface area of CMs before and after CAPP treatment. However, upon subsequent wax extraction, surface area of CMs decreased indicating the release of strain. Increasing the duration of CAPP treatment resulted in increasing strain release up to 47.7 ± 8.0 % at 20 min when CAPP was applied to the inner surface. In contrast, strain release was independent of CAPP duration averaging about 12.1 ± 0.6 % when applied to the outer surface of the CM. Our results provide evidence for a marked gradient of strain between the outer side (strained) and the inner side of the CM (not strained) of mature apple fruit.     

Formulation development of the biocontrol agent Bacillus subtilis strain CPA-8 by spray-drying

Aims: To prepare commercially acceptable formulations of Bacillus subtilis CPA‐8 by spray‐drying with long storage life and retained efficacy to control peach and nectarine brown rot caused by Monilinia spp. Methods and Results: CPA‐8 24‐h‐ and 72‐h‐old cultures were spray dried using 10% skimmed milk, 10% skimmed milk plus 10% MgSO₄, 10% MgSO₄ and 20% MgSO₄ as carriers/protectants. All carriers/protectants gave good percentages of powder recovery (28–38%) and moisture content (7–13%). CPA‐8 survival varied considerably among spray‐dried 24‐h‐ and 72‐h‐old cultures. Seventy‐two hours culture spray dried formulations showed the highest survival (28–32%) with final concentration products of 1·6–3·3 × 10⁹ CFU g^?1, while viability of 24‐h‐old formulations was lower than 1%. Spray‐dried 72‐h‐old formulations were selected to subsequent evaluation. Rehydration of cells with water provided a good recovery of CPA‐8 dried cells, similar to other complex rehydration media tested. Spray‐dried formulations stored at 4 ± 1 and 20 ± 1°C showed good shelf life during 6 months, and viability was maintained or slightly decreased by 0·2–0·3‐log. CPA‐8 formulations after 4‐ and 6 months storage were effective in controlling brown rot caused by Monilinia spp. on nectarines and peaches resulting in a 90–100% reduction in disease incidence. Conclusions: Stable and effective formulations of biocontrol agent B. subtilis CPA‐8 could be obtained by spray‐drying. Significance and Impact of the Study: New shelf‐stable and effective formulations of a biocontrol agent have been obtained by spray‐drying to control brown rot on peach.     

Effect of partial drying and desiccation on somatic seedling quality in Norway and Serbian spruce

Somatic embryo quality is still a problem for many researchers. To improve the efficiency of germination, special procedures are used, such as partial drying of somatic embryos at high relative humidity or desiccation in the presence of supersaturated solutions of salt. In this work, cotyledonary somatic embryos of Norway spruce (Picea abies) and Serbian spruce (P. omorika) were placed on culture media (ME or BM-5) to germinate. We found that after 4 weeks of incubation on these media, hypocotyl and radicle growth of control (non-dried) somatic embryos of both species was not adequate to yield seedlings able to acclimatize to greenhouse conditions. Therefore, somatic embryos were partly dried at relative humidity of 97 % or desiccated at relative humidity of 79 %, for 2 or 3 weeks, and then placed on the Margara (ME) medium. Partial drying of somatic embryos at the higher relative humidity (97 %) enabled an improvement of radicle growth of germinating somatic embryos in both species. The highest conversion rate (45 %) was obtained for embryos of Norway spruce maintained for 2 weeks at relative humidity of 97 %. This treatment contributed to the improvement of germination and conversion efficiency of somatic embryos of Norway spruce, regardless of the drying period. Improved radicle growth facilitated development of better quality seedlings of this spruce species. In Serbian spruce, we did not obtain seedlings of sufficient quality, due to poor hypocotyl growth. Desiccation at humidity of 79 % for 3 weeks proved to be lethal to somatic embryos of both species.     

Cuticular penetration of potassium salts: Effects of humidity,anions, and temperature

Cuticular penetration of potassium salts across astomatous cuticular membranes isolated from pear and Citrus leaves has been studied. Penetration was a first order process and was greatly affected by humidity and hygroscopicity of salts. Temperature did not significantly influence rates of penetration in the range of 10 – 25 °C. Penetration required dissolution of the salt and this is determined by the point of deliquescence (POD) and humidity. When humidity is above the POD the salt residue on the cuticle dissolves, while below a solid residue is formed and penetration ceases. The POD for K₂CO₃ is 44% at 20 °C and above this humidity salt penetration was possible. Rate constants of penetration increased from 0.022 to 0.045 h^–1 when humidity increased from 50 to 90%. With KCl (POD of 86%) rates were highest at 90% and lower at 100% while with KNO₃ (POD of 95%) and KH₂PO₄ (POD of 97%) highest rates were measured at 100% humidity and at 90% penetration was extremely slow. With KCl, KNO₃ and K₂CO₃ maximum rate constants of penetration across pear CM were around 0.04 – 0.05 h^–1 which correspond to half times of penetration of 14 – 17 h. Rate constants measured with Citrus CM were up to two times higher but response to humidity was similar. The potassium salts studied did not only differ in POD and sensitivity to humidity but also in potassium content, which is highest with K₂CO₃ (57%) and KCl (52%) and much lower with KNO₃ (38%) and KH₂PO₄ (29%). Thus, K₂CO₃ is best suited for foliar applications as rates of penetration were large at 50% humidity and higher, while with the other salts humidity must be 90 – 100% for maximum rates of penetration.     

Foliar application of aminoethoxyvinylglycine (AVG) delays fruit ripening and reduces pre-harvest fruit drop and ethylene production of bagged “Kogetsu” apples

Covering apple fruits with double layer waterproof bags to enhance fruit quality and evenness of blush colour is typical on many cultivars in Korea and Japan. Aminoethoxyvinylglycine (AVG) applied to unbagged apple fruits at 3–4 weeks before commercial harvest reduces ethylene production in the fruit, delays fruit ripening and reduces pre-harvest fruit drop. Spray application of AVG to trees of bagged apples should have no effect on apple ripening as there is␣no direct contact with the fruit and the translocation of AVG in apple trees is regarded as negligible. However, preliminary experiments suggested that AVG applied to trees of bagged apples reduced pre-harvest fruit drop in “Kotgetsu” apples. This study investigated the effect of spray treatments of 125 ppm of AVG on fruit drop, fruit ripening (firmness, starch conversion and soluble solids) and ethylene production to whole trees with bagged or unbagged “Kogetsu” fruit, as well as sprays of only the bagged or unbagged fruit on trees on two orchards. AVG applied to whole trees with unbagged apples reduced fruit drop from an average of 58.9% to 10.4%, delayed starch conversion and decreased ethylene production. AVG applied to whole trees with bagged fruit was equally effective in reducing pre-harvest drop, delaying fruit ripening and reducing ethylene production. Application of AVG to unbagged fruit only was nearly as effective as application to whole trees with unbagged fruit but application to bagged fruit only had no effect on fruit ripening or ethylene production. Application of AVG to bagged fruit only did reduce fruit drop to an average of 42.5% but this was not as effective as spraying unbagged fruit only or whole trees with bagged fruit. Possible mechanisms for this effect are discussed.     

Method of testing insecticide susceptibility of Stenotus rubrovittatus (Hemiptera: Miridae) by topical application

Appropriate droplet size and the preferred sex and age of insects for insecticides used in topical application were investigated to standardize the method of testing the susceptibility of Stenotus rubrovittatus (Matsumura). The overflow of acetone droplets applied on the notum of the thorax of S. rubrovittatus adults occurred when the droplet size exceeded 0.20 μl per insect, and the overflow occurred more frequently in males than in females. No effect of acetone solution on mortality was observed at less than 0.20 μl droplet size per insect within 72 h after treatment. The LD₅₀ values and relative potencies of fenitrothion 24 and 48 h after treatment were almost the same in each sex. The fenitrothion susceptibility of males was twice higher than that of females. The fenitrothion susceptibility increased with age after emergence. However, there were small differences in LD₅₀ values and relative potencies between 4–7 and 8–11 days after emergence because the females 4–11 days after emergence had a small difference in body weight. These results led us to conclude that the recommended size of acetone droplets per insect is less than 0.20 μl and that it is suitable to use S. rubrovittatus females 4–11 days after emergence for topical application.     

Effect of air-blast drying and the presence of protectants on the viability of yeast entrapped in calcium alginate beads with an aim to improve the survival rate

Five yeast strains, Saccharomyces cerevisiae D8, M12, and S13; Hanseniaspora uvarum S6; and Issatchenkia orientalis KMBL5774, isolated from Korean grapes, were entrapped in Ca-alginate beads, which are non-toxic, simple to use, and economical. Ca-alginate beads containing yeast cells were soaked in protective solutions, such as skim milk, saccharides, polyols, and nitrogen compounds, before air-blast drying to improve the yeast survival rate and storage ability. The results showed that both entrapment in Ca-alginate beads and soaking in protective agents favorably affected the survival of all strains. The microenvironment formed by the beads and protective agents can protect the yeast cells from harsh environmental conditions, such as low water (below 10 %). All the yeast strains entrapped in Ca-alginate beads showed greater than 80 % survival and less than 11 % water content after air-blast drying at 37 °C for 5 h. In addition, air-blast dried cells of S. cerevisiae D8, M12, S13; H. uvarum S6; and I. orientalis KMBL5774 entrapped in 2 % Ca-alginate beads and soaked in protective agents (10 % skim milk containing 10 % sucrose, 10 % raffinose, 10 % trehalose, 10 % trehalose, and 10 % glucose, respectively) after air-blast drying at 37 °C for 5 h showed 90, 87, 92, 90, and 87 % viability, respectively. All dried entrapped yeast cells showed survival rates of at least 51 % after storage at 4 °C for 3 months.     

Synergistic roles of leaf boron and calcium during the growing season in affecting sugar and starch accumulation in ripening apple fruit

Fruit sugar content is one of the most important flavor quality traits in the fresh market. Minerals, such as boron (B) and calcium (Ca), are associated with fruit sugar and starch accumulation in many plant species. To better understand the roles of B and Ca in affecting sugar and starch accumulation in apples, 2 g L^?1 Na₂B₄O₇·10H₂O or 10 g L^?1 CaCl₂ was supplied by foliar spray to 20-year-old ‘Fuji’ (Malus domestica Borkh. cv. Fuji) trees at four developmental stages (fruit set, onset of rapid fruit growth, rapid fruit growth and the end of rapid fruit growth), in 2010–2011. The most effective treatment significantly increasing soluble sugar and starch levels in ripening fruit was the foliar application of 2 g L^?1 Na₂B₄O₇·10H₂O during rapid fruit growth, and the robustness of the effects was confirmed for two cultivars, ‘Fuji’ and ‘Orin’, at three orchards in 2011. Foliar applications of B during the onset of rapid fruit growth and rapid fruit growth, as well as the foliar application of Ca at fruit set, significantly increased the soluble sugar content in ripening fruit. In addition, the B application was effective in increasing the fruit starch content, but Ca was not. Both B and Ca treatments significantly increased the leaf concentrations of the other element at least transiently. However, B and Ca effects on fruit sugar/starch did not seem to depend on higher leaf B or Ca levels. In conclusion, B and Ca interact in enhancing fruit sugar and starch contents at the fruit ripening stage.     

Yeast Cell Microcapsules as a Novel Carrier for Cholecalciferol Encapsulation: Development,Characterization and Release Properties

In this study Saccharomyces cerevisiae yeast cells was used as a novel vehicle for encapsulation of vitamin D₃. The effects of initial cholecalciferol concentration (100,000 and 500,000 IU/g yeast), yeast cell pretreatment (plasmolysis with NaCl) and drying method (spray or freeze drying) on microcapsules properties were investigated. It was found that the vitamin concentration and drying method had significant influence on encapsulation efficiency (EE) and size of yeast microcapsules. Furthermore, EE values were more increased by the plasmolysis treatment. The highest EE was obtained for plasmolysed and spray dried yeast cells prepared using initial cholecalciferol concentration of 2.5 mg per gram of yeast cells (76.10?±?6.92%). The values of mean particle size were 3.43–7.91 μm. The presence of cholecalciferol in yeast microcapsules was confirmed by X-ray diffraction (XRD) and Fourier transform-infrared (FT-IR) analyses. The in vitro cholecalciferol release from yeast microcapsules in phosphate buffer saline solution (PBS) followed a controlled release manner consistent with a Fickian diffusion mechanism. In addition, the release studies in simulated gastrointestinal tract showed sustained release of cholecalciferol in the stomach condition and significant release in intestinal medium.     

Combined effects of spray‐drying conditions and postdrying storage time and temperature on Salmonella choleraesuis and Salmonella typhimurium survival when inoculated in liquid porcine plasma

The objective of this study was to determine the effectiveness of the spray‐drying process on the inactivation of Salmonella choleraesuis and Salmonella typhimurium spiked in liquid porcine plasma and to test the additive effect of immediate postdrying storage. Commercial spray‐dried porcine plasma was sterilized by irradiation and then reconstituted (1:9) with sterile water. Aliquots of reconstituted plasma were inoculated with either S. choleraesuis or S. typhimurium, subjected to spray‐drying at an inlet temperature of 200°C and an outlet temperature of either 71 or 80°C, and each spray‐drying temperature combinations were subjected to either 0, 30 or 60 s of residence time (RT) as a simulation of residence time typical of commercial dryers. Spray‐dried samples were stored at either 4·0 ± 3·0°C or 23·0 ± 0·3°C for 15 days. Bacterial counts of each Salmonella spp., were completed for all samples. For both Salmonella spp., spray‐drying at both outlet temperatures reduced bacterial counts about 3 logs at RT 0 s, while there was about a 5·5 log reduction at RT 60 s. Storage of all dried samples at either 4·0 ± 3·0°C or 23·0 ± 0·3°C for 15 days eliminate all detectable bacterial counts of both Salmonella spp.

Significance and Impact of the Study

Safety of raw materials from animal origin like spray‐dried porcine plasma (SDPP) may be a concern for the swine industry. Spray‐drying process and postdrying storage are good inactivation steps to reduce the bacterial load of Salmonella choleraesuis and Salmonella typhimurium. For both Salmonella spp., spray‐drying at 71°C or 80°C outlet temperatures reduced bacterial counts about 3 log at residence time (RT) 0 s, while there was about a 5.5 log reduction at RT 60 s. Storage of all dried samples at either 4.0 ± 3.0°C or 23.0 ± 0.3°C for 15 days was effective for eliminating detectable bacterial counts of both Salmonella spp.     

Viability of rape (Brassica napus L.) seeds following selection on the basis of newly-emerged radicles then subsequent drying and storage

Germinating rape seeds selected on the basis of newly-emerged radicles (1 ± 0.5 mm) were dried to an equilibrium moisture content (c. 11%) in air at 20°C and 80% relative humidity without loss of viability. Storage life of these low-moisture-content germinating (LMCG) seeds at 15°C was limited to 7 days before viability was significantly reduced. However, viability of LMCG seeds was maintained for 84 days in storage at -20°C. Longer periods in store reduced viability, but 96% of seeds still remained viable after 336 days at - 20°C. Increasing periods of storage at -20°C reduced the subsequent seed longevity at 15°C, indicating a reduction in vigour during storage. Storage under reduced pressure or in a nitrogen atmosphere had little significant effect on seed longevity. Reduction of moisture content below 11% using vacuum drying at a range of temperatures reduced seed vigour.     

EFFECTS OF YEAST,FERMENTATION TIME,AND PRESERVATION METHODS ON TARHANA

The physicochemical properties of tarhana soup produced with different dough treatments, fermentation times, and preservation methods were examined. Tarhana doughs were prepared with yogurt (control) or baker's yeast (Saccharomyces cerevisiae) and fermented for 3 days. Samples were taken at 24, 48, and 72 hr. Samples were then preserved via one of four methods: sun dried, dried in the shade, vacumn dried, and frozen. Frozen samples produced lower organic acid levels after 72 hr of fermentation in both control (0.68 g/100 g) and yeast (0.61 g/100 g) applications than samples that were dried (0.94 g/100 g control samples; 0.81 g/100 g samples with yeast). Increasing fermentation time resulted in a significant effect on the formation of organic acid in the tarhana (p < .01). At 72 hr of fermentation, total acidity increased 11%, 17%, and 23% for tarhana samples vacumn-dried, sun-dried, and dried in the shade, respectively. Preservation methods also affected the moisture, ash, crude protein, total acidity, pH, salt, fat, reducing sugar levels, and the sensory assestment of tarhana soup (p < .01). Sensory characteristics were not significantly affected by baker's yeast in any of the preservation methods used (p > .01). However, sensory scores for tarhana prepared from the samples dried in a sheltered area showed a reduction in color desireablilty as the fermentation time increased. The soup prepared from frozen tarhana (72 hr fermentation, with yeast) had the highest scores with respect to color, mouth feel, flavor, and overall acceptability. Vacuum-dried samples' scores in these areas were also high in comparison to the two other drying methods.     

Batch anaerobic co-digestion of cow manure and waste milk in two-stage process for hydrogen and methane productions

Anaerobic co-digestion of cow manure (CM) and waste milk (WM), produced by sick cows during treatment with antibiotics, was evaluated in two-stage process under thermophilic condition (55 °C) to determine the effect of WM addition on hydrogen (H₂) and methane (CH₄) production potentials, volatile solids (VS) removal, and energy recovery. Six CM to WM VS ratios of 100:0, 90:10, 70:30, 50:50, 30:70, and 10:90 were examined using 1-L batch digesters. The WM VS ratio of 30 % was found to be the minimum limit for significant increases in specific H₂ and CH₄ yields, and VS removal as compared to digestion of manure alone (P < 0.05). The highest specific H₂ and CH₄ yields, VS removal and energy yield were 38.2 mL/g VS, 627.6 mL/g VS, 78.4 % and 25,459.8 kJ/kg VS, respectively, in CM:WM 30:70. Lag phases to H₂ and CH₄ productions were observed in CM–WM mixtures, increased with increasing the amount of WM in the feedstock and were greater than 72 h in CM:WM 50:50 and 30:70. The digestion system failed in CM:WM 10:90. The results suggest that CM:WM 30:70 was optimum, however, due to limited amount of WM usually generated and long lag phase at this ratio which may make the process uneconomical, CM:WM 70:30 is recommended in practice.     

Formulation of the biocontrol agent <Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> CPA-8 using different approaches: liquid,freeze-drying and fluid-bed spray-drying

The present work focuses on the assessment and comparison of three different formulation technologies and the effect of protectants on cell viability, storage stability and antagonistic activity of the biocontrol agent Bacillus amyloliquefaciens CPA-8. Cultures were concentrated with different protective substances such as MgSO₄, sucrose and skimmed milk (SM) and subjected to liquid formulation, freeze-drying and fluid-bed spray-drying. Results showed that CPA-8 freeze-dried cells without protectants or amended with SM suffered the highest losses in cell viability (0.41?0.48 log). Moreover, the cell viability of the tested freeze-dried products decreased after four months of storage at both tested temperatures (4 and 22 °C). Otherwise, liquid and fluid-bed spray-dried products were stable for four months at 4 °C and for 12 months at 22, 4 and ?20 °C, respectively, and no effect of the protectants was observed. The most suitable CPA-8 products were then tested against Monilinia laxa and M. fructicola in artificially wounded nectarines and in all cases the antagonistic activity was maintained similar to fresh cells. The efficacy results revealed that the formulation process did not affect the biocontrol potential of CPA-8. This work led us to conclude that effective formulations with final concentrations ranging from 1.93 × 10⁹–2.98 × 10⁹ CFU ml^?1 and from 4.76 × 10⁹–1.03 × 10¹⁰ CFU g^?1 were obtained for liquid and dried products, respectively. Additionally, the suitability of the fluid-bed spray drying technology should be taken into account to develop a stable and effective CPA-8 product for practical applications to control brown rot in stone fruit.     

Investigating Gabapentin Polymorphism Using Solid-State NMR Spectroscopy

Solid-state NMR spectroscopy (SSNMR), coupled with powder X-ray diffraction (PXRD), was used to identify the physical forms of gabapentin in samples prepared by recrystallization, spray drying, dehydration, and milling. Four different crystalline forms of gabapentin were observed: form I, a monohydrate, form II, the most stable at ambient conditions, form III, produced by either recrystallization or milling, and an isomorphous desolvate produced from desolvating the monohydrate. As-received gabapentin (form II) was ball-milled for 45 min in both the presence and absence of hydroxypropylcellulose (HPC). The samples were then stored for 2 days at 50°C under 0% relative humidity and analyzed by ¹³C SSNMR and PXRD. High-performance liquid chromatography was run on the samples to determine the amount of degradation product formed before and after storage. The ¹H T ₁ values measured for the sample varied from 130 s for the as-received unstressed material without HPC to 11 s for the material that had been ball-milled in the presence of HPC. Samples with longer ¹H T ₁ values were substantially more stable than samples that had shorter T ₁ values. Samples milled with HPC had detectable form III crystals as well. These results suggest that SSNMR can be used to predict gabapentin stability in formulated products.     

Foliar application of plant nutrients and kinetin modifies growth and essential oil profile in <Emphasis Type="Italic">Rosa damascena</Emphasis> under acidic conditions

Rosa damascena Mill. is cultivated for its high-value essential oil in different parts of the world. The flower yield and the composition of essential oil of R. damascena are strongly affected by a number of factors. Nevertheless, the interactive effects of foliar application of plant nutrients and kinetin and its time of application on yield and secondary metabolites profile of R. damascena under acidic conditions are still unclear. Thus, a field experiment comprising two different times of spray and five foliar spray treatments was conducted to test the hypothesis that flowering behavior and secondary metabolites profile can be modified through proper nutrient supply at right time. The foliar spray at flower bud appearance stage (S₂) significantly (P ≤ 0.05) increased flower yield by about 10.0 % compared with the foliar application at axillary bud development stage (S₁) during both years, regardless of plant nutrients. Among the foliar spray treatments, kinetin at 0.20 g L^?1 registered about 23–39 % higher flower yield compared with the water spray control; however, remained statistically at par (P ≤ 0.05) with Ca(NO₃)₂ at 4.06 g L^?1. Moreover, the percentage of major fragrance-bearing compounds of essential oil (β-citronellol + nerol, linalool, E-geraniol, and Z-citral) was marginally increased with Ca(NO₃)₂ compared with kinetin treatment. However, the percentages of major hydrocarbons, nonadecane and heneicosane, were noticeably increased when kinetin was applied at S₁. Foliar application of kinetin and Ca(NO₃)₂ might be done to improve flower yield and essential oil content in R. damascena flowers.     

Gibberellins Increase Cuticle Deposition in Developing Tomato Fruit

Effects of the gibberellins A₄₊₇(GA₄₊₇) and A₃(GA₃), benzyladenine (BA) and forchlorfenuron (CPPU) on deposition of the cuticular membrane (CM) in developing tomato (Lycopersicon esculentum L.) fruit were investigated. Growth regulators were applied when fruit development within trusses ranged from the flower to the mature stage. Developmental stage of fruit at the time of application was indexed by fruit diameter. Fruit were harvested at maturity, the CM isolated enzymatically on an individual fruit basis and mass of CM per unit fruit surface area calculated. In mature fruit, mass of CM per fruit increased with fruit size, but mass of CM per unit surface area was independent of fruit size, position within a truss and position of the truss on the plant. GA₄₊₇ and GA₃ increased CM mass per unit fruit surface area at concentrations up to 300 mg l⁻¹. Young fruit (5–10 mm diam. at time of application) was most responsive. Responsiveness decreased as fruit development at application progressed towards maturity. There was no consistent effect of GA₄₊₇ or GA₃ on fruit mass. BA (up to 100 mg l⁻¹) or CPPU (up to 3 mg l⁻¹) had no significant effect on CM mass per unit surface area regardless of developmental stage. Higher concentrations of BA or CPPU decreased CM mass per unit surface area. There was no effect of BA or CPPU on fruit mass. Potential mechanisms and benefits of a gibberellin induced increase in CM deposition are discussed.     

Reduced late-season leaf potassium and phosphorus levels influence decreases in sugar contents of bagged apple fruit

Fruit bagging has been widely used in the fruit production industry; however, in apples, it is known to cause a significant decrease in fruit sugar contents. To address this issue and identify the changes in leaf mineral contents associated with fruit sugar levels in bagged apples, aqueous solutions of 10 g L^?1 CaCl₂, 5 g L^?1 KH₂PO₄, or 2 g L^?1 Na₂B₄O₇·10H₂O were foliar sprayed during four fruit developmental stages. The late-season leaf phosphorus (P) and potassium (K) contents after the rapid fruit growth period and the soluble sugar contents in ripening fruit were significantly lower in bagged fruit than in non-bagged fruit (11.1–15.09 %). The decreases in leaf P and K contents caused by bagging were almost completely compensated for by foliage applications of CaCl₂, KH₂PO₄, or Na₂B₄O₇ during the fruit set period. Therefore, the fruit soluble sugar contents were significantly higher in bagged ripening fruit with foliar spray than in bagged fruit without foliar sprays, reaching the levels of non-bagged apples. The decrease in the sugar contents of bagged apples was closely associated with the decrease in late-season leaf P and K levels caused by fruit bagging.     

Investigation of the antibacterial activity of a short cationic peptide against multidrug-resistant Klebsiella pneumoniae and Salmonella typhimurium strains and its cytotoxicity on eukaryotic cells

With the growing microbial resistance to conventional antimicrobial agents, the development of novel and alternative therapeutic strategies are vital. During recent years novel peptide antibiotics with broad spectrum activity against many Gram-positive and Gram-negative bacteria have been developed. In this study, antibacterial activity of CM11 peptide (WKLFKKILKVL-NH2), a short cecropin–melittin hybrid peptide, is evaluated against antibiotic-resistant strains of Klebsiella pneumoniae and Salmonella typhimurium as two important pathogenic bacteria. To appraise the antibacterial activity, minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC) and bactericidal killing assay were utilized with different concentrations (2–128 mg/L) of peptide. To evaluate cytotoxic effect of peptide, viability of RAJI, Hela, SP2/0, CHO, LNCAP cell lines and primary murine macrophage cells were also investigated with MTT assay in different concentrations (3–24 and 0.5–16 mg/L, respectively). MICs of K. pneumoniae and S. typhimurium isolates were in range of 8–16 and 4–16 mg/L, respectively. In bactericidal killing assay no colonies were observed at 2X MIC for K. pneumoniae and S. typhimurium isolates after 80–90 min, respectively. Despite the fact that CM11 reveals no significant cytotoxicity on RAJI, Hela, SP2/0, and CHO cell lines beneath 6 mg/L at first 24 and 48 h, the viability of LNCAP cells are about 50 % at 3 mg/L, which indicates strong cytotoxicity of the peptide. In addition, macrophage toxicity by MTT assay showed that LD₅₀ of CM11 peptide is 12 μM (16 mg/L) after 48 h while in this concentration after 24 h macrophage viability was about 70 %.     

Mannitol-enhanced survival of Lactococcus lactis subjected to drying

Survival of Lactococcus lactis subjected to different drying conditions was investigated. Mannitol most remarkably enhanced the survival of dried cells to a level almost equalling that of viable cells [log₁₀ (cfu ml⁻¹) = 9.42] as was found prior to the drying process (log₁₀ = 9.6). In the absence of mannitol, a survival was reduced by a factor of 10⁴. Drying of cells at 20 °C led to higher survival rates than drying at 30 °C. Mannitol enhanced the survival rate at both temperatures, and at both 20 °C and 30 °C the highest reduction in survival occurred when cells were dried at a water activity of 0.76. In the presence of mannitol, differences in survival after drying at different water activities were less pronounced. Rehydration of cells dried in the presence of mannitol resulted in an extended lag phase of 4 h compared to fresh cells. No growth or acidification of the culture medium was observed for 12 h in the case of rehydrated cells dried in the absence of mannitol. It was hypothesized that a radical scavenging activity of mannitol could partly explain these observations. Received: 28 August 1998 / Accepted: 2 October 1998