Simultaneous and sensitive detection of three foodborne pathogens by multiplex PCR, capillary gel electrophoresis, and laser-induced fluorescence

The simultaneous detection of Staphylococcus aureus, Listeria monocytogenes, and Salmonella spp. has been approached by a new multiplex PCR-based procedure followed by capillary gel electrophoresis with laser-induced fluorescence detection (multiplex-PCR-CGE-LIF). As compared to slab gel electrophoresis, the use of CGE-LIF improved from 10- to 1000-fold the sensitivity of the multiplex PCR analysis, allowing the detection of 2.6 x 10(3) cfu mL(-1) of S. aureus, 570 cfu mL(-1) of L. monocytogenes, and 790 cfu mL(-1) of Salmonella in artificially inoculated food, without enrichment. Following 6 h of enrichment, as low as 260, 79, and 57 cfu mL(-1) of S. aureus, L. monocytogenes, and Salmonella, respectively, were detected. The CGE-LIF method is shown to be reproducible, providing relative standard deviation (RSD) values lower than 0.8% for analysis time and lower than 5.8% for peak areas. The multiplex-PCR-CGE-LIF proved a powerful analytical tool to detect various food pathogens simultaneously in a fast, reproducible, and sensitive way.     

Micropropagation of <Emphasis Type="Italic">Salvadora</Emphasis><Emphasis Type="Italic">persica</Emphasis>-a tree of arid horticulture and forestry

A micropropagation method for Jaal (Salvadora persica)—a tree of arid horticulture and forestry has been developed. Nodal segments of fresh shoot sprouts originated from axillary buds obtained from a plant around 35–40 years old lopped plant were used as explants for establishment of in vitro cultures. Surface-sterilized explants produced optimum number of shoots through activation of axillary buds on Murashige and Skoog’s (MS) medium containing 8.88 μM BA (6-benzyladenine) + additives (25 mgl⁻¹ each of adenine sulphate, arginine, citric acid, 50 mgl⁻¹ ascorbic acid). The shoot multiplication was influenced by the successive transfer of the mother explants for 4–5 passages. The maximum number (23.1 ± 0.73 shoots per explant) of shoots were regenerated on MS supplemented with 1.11 μM BA + 1.16 μM Kn (Kinetin) + 0.54 μM NAA (α-naphthalene acetic acid). About 90% shoots pulse-treated with a combination of 2460.27 μM Indole-3-butyric acid (IBA) + 494.56 μM NOA (2-naphthoxy acetic acid) were rooted ex vitro on soilrite within 15–18 days. Over 80% cloned plantlets were hardened successfully in a green house and transferred to polybag/pots.     

Soil biological properties under different tree based traditional agroforestry systems in a semi-arid region of Rajasthan,India

An investigation was carried out in an Entisol at farmers’ field in Jaipur district, Rajasthan, India during 2002–2004 to evaluate the effect of traditionally grown trees on soil biological characteristics. Traditionally grown trees in farm lands for study consisted of Prosopis cineraria (L.), Dalbergia sissoo (Roxb.) ex DC, Acacia leucophloea (Roxb.) and Acacia nilotica (L.) Del. having a canopy diameter of 8 m. Results revealed significant and substantial improvement in soil biological activity in terms of microbial biomass C, N and P, dehydrogenase and alkaline phosphatase activity under different tree based agroforestry systems as compared to a no tree control (cropping alone). Soil microbial biomass C, N and P under agroforestry varied between 262–320, 32.1–42.4 and 11.6–15.6 μg g⁻¹ soil, respectively, with corresponding microbial biomass C, N and P of 186, 23.2 and 8.4 μg g⁻¹ soil under a no tree control. Fluxes of C, N and P through microbial biomass were also significantly higher in P. cineraria based land use system followed by D. sissoo, A. leucophloea and Acacia nilotica in comparison to a no tree control. Thus, it is concluded that agroforestry system at farmers’ field enhance soil biological activity and amongst trees, P. cineraria based system brought maximum and significant improvement in soil biological activity.     

Effects of rainwater harvesting on herbage diversity and productivity in degraded Aravalli hills in western India

Over-exploitation and rural growth have severely damaged native vegetations of Aravalli hills in Rajasthan, India. This study was conducted to evaluate the effects of different restoration practices (i.e., rainwater harvesting (RWH) and planting of tree seedlings) on improve- ment in soil water and nutrients and growth and biomass of herbaceous vegetation. Contour trench (CT), Gradonie (G), Box trench (BT), V-ditch (VD) and a control were imposed on 75 plots (each of 700 m 2 ) in natural slope gradient defined as <10%, 10% 20% and >20% slopes in 2005. Each plot had three micro-sites of 1-m 2 at up (USP), middle (MSP) and lower (LSP) part of the plot for observation in 2008. The existed gradient (due to soil texture and topographic features) of soil pH, EC, SOC, NH 4 - N, NO 3 -N and PO 4 -P in June 2005 between >20% to <10% slopes were decreased in 2008 after applying RWH techniques. Such improvement in soil status promoted vegetation growth and biomass in higher slope gra- dients. Soil water, species diversity and herbage biomass increased from USP to LSP, and RWH techniques had positive role in improving SOC, nutrients, vegetation population, evenness and growth at MSP. Despite of lowest SWC, regular rain and greater soil water usage enhanced green and dry herbage biomasses in 10% 20% and >20% slopes, compared with <10% slope. The highest diversity in CT treatment was related to herbage biomass, which was enhanced further by highest concentrations of SOC and PO 4 -P. Further, CT treatment was found to be the best treat- ment in minimizing biomass variance in different slopes. Conclusively, soil texture and topographic features controlled soil water and nutrients availability. Rainwater harvesting techniques increased soil water storage and nutrient retention and also enhanced vegetation status and biomass by minimizing the effects of hillslopes. Thus depending upon the site conditions, suitable RWH technique could be adopted to increase herb- age biomass while rehabilitating the degraded hills.     

Characterization of the interaction between two food aroma components, alpha-pinene and ethyl butyrate, and ethylene-vinyl alcohol copolymer (EVOH) packaging films as a function of environmental humidity

The ethylene-vinyl alcohol copolymers (EVOHs) are well-known high oxygen barrier materials that are being used successfully in the design of packaging structures for oxygen-sensitive food or pharmaceutical products. Recently, there has been increasing interest in using EVOH materials to provide a high barrier to organic compounds as a means to reduce food aroma scalping. However, the barrier function of this family of materials diminishes significantly in humid environments, and it is supposed that so does the organic vapor barrier. In this work, a new sorption-based method to characterize the interaction between food aroma and polymer films for packaging as a function of relative humidity is presented and is used to determine the barrier to ethyl butyrate and alpha-pinene of EVOH at 23 degrees C. The results show that although EVOH is an excellent barrier to food aroma when dry, a property that even improves at low relative humidity (RH), the solubility and diffusivity of the compounds tested increase dramatically with humidity at medium to high water activities. However, even in the worst case (100% RH), EVOH outperforms low-density polyethylene (LDPE) as a barrier to organic vapors at least 500,000-fold.     

Effect of accelerated autolysis of yeast on the composition and foaming properties of sparkling wines elaborated by a champenoise method

Five mutants (obtained by UV mutagenesis) and the parent strain were selected to produce sparkling wines following the traditional or champenoise method. The wines were aged with the yeast for 9 months, with samples being taken each month for analytical and sensory determinations. The wines elaborated with mutant strain IFI473I demonstrated an accelerated release of protein, amino acids, and polysaccharides. An analysis of the secreted polysaccharides revealed that mannose was the major sugar present. The effects of the products released by yeasts on the foaming properties of the wines were determined by both sensory and instrumental analysis. In all cases, the wines elaborated with mutant strain IFI473I showed improved foaming properties as compared to wines fermented without this strain. Similar results were obtained at a decreased aging time of 6 months, thereby confirming the capacity of IFI473I strain to carry out an accelerated autolysis. These results demonstrate that mutant strain IFI473I can significantly reduce production times of high-quality sparkling wines.     

Chemometric studies of vinegars from different raw materials and processes of production

The phenolic composition, aroma compounds, and organic acid content of 83 vinegars have been determined. Multivariate analysis techniques have been used to classify these vinegar samples according to raw material (white wine, red wine, apple, honey, alcohol, balsamic, and malt) and production process (with and without aging in wood). Cluster analysis grouped the samples according to production process. Only apple and balsamic vinegars were separated from wine vinegars. Alcohol, honey, and malt vinegars were grouped with no aged wine vinegars. Linear discriminate analysis allowed a 88% differentiation according to raw material and 100% according to aging in wood. Besides, from the results obtained, a major role of the volatile compounds in the differentiation of the vinegar samples according to their aging period in wood can be seen.     

Varietal tolerance to Zinc deficiency in wheat and barley grown in chelatorbuffered nutrient solution and its effect on uptake of Cu,Fe, and Mn

Tolerance to zinc (Zn) deficiency was examined for three wheat (Triticum aestivum L.) and three barley (Hordeum vulgare L.) varieties grown in chelator‐buffered nutrient solution. Four indices were chosen to characterize tolerance to Zn deficiency: (1) relative shoot weight at low compared to high Zn supply (“Zn efficiency index”), (2) relative shoot to root ratio at low compared to high Zn supply, (3) total shoot uptake of Zn under deficient conditions, and (4) shoot dry weight under deficient conditions. Barley and wheat exhibited different tolerance to Zn deficiency, with barley being consistently more tolerant than wheat as assessed by all four indices. The tolerance to Zn deficiency in the barley varieties was in the order Thule=Tyra>Kinnan, and that of wheat in the order Bastian=Avle>Vinjett. The less tolerant varieties of both species accumulated more P in the shoots than the more tolerant varieties. For all varieties, the concentrations of Mn, Fe, Cu, and P in shoot tissue were negatively correlated with Zn supply. This antagonism was more pronounced for Mn and P than for Cu and Fe. Accumulation of Cu in barley roots was extremely high under Zn‐deficient conditions, an effect not so clearly indicated in wheat.     

Effect of different forms and sources of arsenic on crop yield and arsenic concentration

Two greenhouse experiments were conducted to investigate the effects of different sources of As application on the yield of ryegrass (Lolium perenne L.) and barley (Hordeum vulgare L.) and the arsenic concentration in crop tissue. The soils used were a sand and a loam soil. In the first experiment (3 yr), As was applied either as sodium arsenite or as disodium hydrogen arsenate at the rate of 0, 2, 10, 50, and 250 mg As kg^?1 soil in year 1. In years 2 and 3, only the residual effect of As applied in year 1 was investigated. In the second experiment (5 yr), a NPK(16-7-12) fertilizer containing 10, 100 and 3000 mg As kg^?1 as sodium arsenite was applied at rates of 750 and 600 mg kg^?1 to ryegrass and barley, respectively, with lime and without lime. Arsenite had more inhibitory effect on crop yield than arsenate. Although the residual effect persisted up to year 3, it was found to be dependent on As application rate, soil type, and crop species. Only minor differences in As concentration in crops between arsenite and arsenate sources were observed. The yield reduction and the increase of As concentration in crops was highly related to As application rates. The As rates of 50 and 250 mg As kg^?1 resulted in significant yield reduction and marked increase of As concentration in both crops and these rates could be considered as critical for both crops. The application of As containing fertilizer had little or no effect on crop yield. The As concentration in both crops was little affected by the fertilizer containing up to 100 mg As kg^?1 but the concentration increased several folds when the fertilizer containing 3000 mg As kg^?1 was applied. The application of additional P to the treatment containing 3000 mg As kg^?1 fertilizer only slightly increased crop yields but it increased As concentration in ryegrass markedly. The repeated application of As containing fertilizer up to a period of 5 yr, even at 3000 mg As kg^?1 level, generally did not affect either the yields or the As concentration in both crops although a slight increase of As concentration in barley grain over the years was observed. In both experiments, the yield reduction and the increase in As concentration in crop tissue were lower in the loam soil than in the sand and barley straw cointained much higher As than barley grains, indicating that greater proportion of the As taken up by this crop was retained in the vegetative organs.     

High frequency plantlet regeneration from nodal segment culture of female <Emphasis Type="Italic">Momordica dioica</Emphasis> (Roxb.)

An in vitro propagation method for female plants of Momordica dioica (Roxb.) has been established. The nodal segments were harvested and the cut ends of the explants were sealed with wax and then surface sterilized and cultured. Bud breaking occurred on Murashige and Skoog’s (MS) agar-gelled medium + 2.0 mg L⁻¹ 6-Benzylaminopurine (BAP) + 0.1 mg L⁻¹ Indole-3 acetic acid (IAA). The cultures were amplified by passages on MS medium supplemented with 1.0 mg L⁻¹ BAP + 0.1 mg L⁻¹ IAA. Further, shoot amplification (29.2 shoots per vessel) was achieved by subculturing of in vitro regenerated shoot clump on MS medium + 0.5 mg L⁻¹ BAP + 0.1 mg L⁻¹ IAA. The micropropagated shoots were subsequently transferred for root formation on half-strength MS medium + 2.0 mg L⁻¹ Indole-3 butyric acid (IBA) with 89% success rate. The in vitro-regenerated shoots were also rooted ex vitro with 34% success. These plantlets were hardened in the greenhouse and transferred to the field. The established protocol is suitable for true to type cloning of mature female plant of M. dioica.