Analysis of volatiles in porcine liver pâtés with added sage and rosemary essential oils by using SPME-GC-MS

The effect of the addition of two natural antioxidant extracts (sage and rosemary essential oils) and one synthetic (BHT) on the generation of volatile compounds in liver patés from Iberian and white pigs was analyzed using SPME-GC-MS. Lipid-derived volatiles such as aldehydes [hexanal, octanal, nonanal, hept-(Z)-4-enal, oct-(E)-2-enal, non-(Z)-2-enal, dec-(E)-2-enal, deca-(E,Z)-2,4-dienal] and alcohols (pentan-1-ol, hexan-1-ol, oct-1-en-3-ol) were the most abundant compounds in the headspace of porcine liver patés. Patés from different pig breeds presented different volatiles profiles due to their different oxidation susceptibilities as a probable result of their fatty acid profiles and vitamin E content. Regardless of the origin of the patés, the addition of BHT successfully reduced the amount of volatiles derived from PUFA oxidation. Added essential oils showed a different effect on the generation of volatiles whether they were added in patés from Iberian or white pigs because they inhibited lipid oxidation in the former and enhanced oxidative instability in the latter. SPME successfully allowed the isolation and analysis of 41 volatile terpenes from patés with added sage and rosemary essential oils including alpha-pinene, beta-myrcene, 1-limonene, (E)-caryophyllene, linalool, camphor, and 1,8-cineole, which might contribute to the aroma characteristics of liver patés.     

Ultrasensitive detection of genetically modified maize DNA by capillary gel electrophoresis with laser-induced fluorescence using different fluorescent intercalating dyes

In this work, four different fluorescent intercalating dyes are compared for the ultrasensitive CGE-LIF detection of DNA from transgenic maize in flours. The fluorescent intercalating dyes compared are YOPRO-1, SYBR-Green-I, Ethidium bromide (EthBr), and EnhanCE. For all the four dyes optimum concentrations are established, and efficient separations of DNA fragments ranging in size from 80 to 1000 bp are obtained. The comparative study demonstrates that SYBR-Green-I and YOPRO-1 provide better limits of detection (LODs) than EnhanCE or EthBr (i.e., LODs are, respectively, 700, 1000, 11300, and 97400 zmol, calculated for a 200-bp DNA fragment). Separations using YOPRO-1 are faster than those using SYBR-Green-I (30 min vs 47 min for the analysis of the 80-1000 bp DNA fragments). Also, separations using YOPRO-1 are more efficient than those using SYBR-Green-I (e.g., 2.4 x 10(6) plates/m vs 1.6 x 10(6) plates/m, respectively, calculated for the 200-bp fragment). Also, buffer depletion and cost per analysis are worse with SYBR-Green-I than with YOPRO-1. Therefore, YOPRO-1 was selected as the preferred intercalating dye. Using this fluorescent compound, analysis time reproducibility for the CGE-LIF separation of the DNA fragments is determined to be better than 1.7% (% RSD, n = 10) within the same day, and better than 1.9% (% RSD, n = 30) for three different days. Moreover, the fluorescence signal obtained using this dye is shown to vary linearly with the DNA concentration in the range studied, i.e., 1-500 ng/microL. It is demonstrated that by using this method 0.01% of transgenic maize can be detected in flour by direct injection of the PCR-amplified sample.     

Quantitative Measurement of Organic Carbon in Mine Wastes: Methods Comparison for Inorganic Carbon Removal and Organic Carbon Recovery

The present study aims to evaluate carbon recovery and analytical precision of two methods based on dry combustion for organic carbon (OC) quantification in base metal mine tailings: (1) IC subtraction method (inorganic C(IC) measured and subtracted from total C) and (2) direct OC quantification after acid pretreatment. Results showed IC subtraction method effectively hydrolyzed a range of IC minerals [calcium carbonate (CaCO₃), magnesium carbonate (MgCO₃), dolomite, magnesite, calcite] with satisfactory IC (as CO₂ released) recovery (87–103 percent). In the direct OC quantification method, 5 M hydrochloric acid (HCl) pretreatment resulted in a satisfactory recovery (76–92 percent) of most organic compounds [ethylenediaminetetraacetic acid (EDTA), cellulose, plant litter, and charcoal], except for water-soluble OC (sucrose, 40 percent recovery). The precision of both methods declined when C levels were <5 g kg^?1 with RSD (Relative Standard Deviation) > 10 percent. The OC values in test samples of base metal mine tailings were comparable between the two methods. However, IC subtraction method is not applicable for tailings with low OC levels (<5 g kg^?1) until the precision is substantially improved. Moreover, compared to the IC subtraction method, OC values are significantly lower in direct OC quantification method for tailings with high OC levels.     

Effect of dietary supplementation of Bacillus licheniformis on gut microbiota, growth and immune response in giant freshwater prawn, Macrobrachium rosenbergii (de Man, 1879)

The present study was conducted to evaluate the effect of dietary supplementation of probiotic bacterium Bacillus licheniformis on the gut microbiota, growth and immune response in Macrobrachium rosenbergii juveniles (4.0 ± 0.02 g). Five isocaloric experimental diets supplemented with different concentrations of B. licheniformis, viz. Control (without probiotic supplementation), T₁ (1.0 × 10⁶ cfu g^?1feed), T₂ (1.0 × 10⁷ cfu g^?1feed), T₃ (1.0 × 10⁸ cfu g^?1 feed) and T₄ (1.0 × 10⁹ cfu g^?1 feed) were used for the experiment. After 60 days of feeding trial, the growth performance increases significantly in all the experimental groups compared with control, and highest weight gain, specific growth rate and lowest feed conversion ratio were recorded from T₄ (P > 0.05). However, juvenile survival was not found to be significantly affected by probiotic supplementation (P > 0.05). The total bacterial count and Bacillus spp. count in the gut were significantly increased with concurrent decrease in pathogenic Aeromonas spp. and Pseudomonas spp. in all the experimental groups compared with control (P < 0.05). Compared with control, immune parameters were increased significantly in all the experimental groups (P < 0.05). Highest haemocyte count, phenol oxidase, superoxide dismutase and antibacterial activity in haemolymph were recorded from T₄ (P < 0.05). When prawns were challenged with Vibrio alginolyticus, the cumulative mortality was significantly lower in T₄ (P < 0.05). These results indicate that dietary supplementation of B. licheniformis can manipulate the gut microbiota and increase the growth and immune response of M. rosenbergii. In the present study, concentration response on the probiotic effect of B. licheniformis was observed which showed higher effects at a concentration of 1.0 × 10⁹ cfu g^?1 feed in the laboratory condition.     

Remarks on genetic diversity and relationship of <Emphasis Type="Italic">Punica protopunica</Emphasis> and <Emphasis Type="Italic">P. granatum</Emphasis> assessed by molecular analyses

Here, two Punica species, viz., P. protopunica Balf. fil., reported as native to Socotra, and P. granatum L., were compared for the first time. Analysis of one P. protopunica and eleven P. granatum accessions was performed using three molecular markers, i.e., sequence related amplified polymorphism (SRAP), target region amplification polymorphism (TRAP), and intron targeted amplified polymorphism (ITAP), along with analysis of pgWD40 sequences, a gene involved in anthocyanin biosynthesis. All markers revealed the relationship between the two species and placed them at 33% similarity. SRAP, TRAP, and ITAP generated a total of 299, 260, and 160 bands, respectively. Of these, 78, 74, and 41 bands were specific for P. protopunica, and 92, 85, and 57 bands, respectively, were shared between both species. Sequence analysis of pgWD40~870 bp amplicons exhibited 100% identity among P. granatum accessions and 98% identity to that of P. protopunica. Phylogenetic analysis of WD40 sequences from monocot and dicot species, including both Punica species confirmed the relation between P. protopunica and P. granatum, supporting earlier reports that P. protopunica could be an ancestral species of P. granatum. Furthermore, the genetic diversity among and within P. granatum accessions from Egypt (3), Mexico (5), and Yemen (3) was assessed. Molecular marker-based relationships among region-bulked accessions was approximately the same (~90% similarity), whereas the degree of genetic variation was altered within each region. Specific bands (alleles) for accessions of each region along with those shared among them were identified. Thus, these bands could be used for pomegranate genotyping and breeding programs.     

Isolation and characterization of immortalized porcine aortic endothelial cell lines

Primary porcine endothelial cells have a limited life span in culture. After four to five passages, they tend to de-differentiate and eventually reach senescence. The aim of this work was to establish immortalized porcine aortic endothelial cell lines (AOCs) to facilitate in vitro studies of different pathological process involving the endothelium. Primary porcine aortic endothelial cells (PAECs) were transfected with a plasmid containing the SV40 genome and selected on the basis of morphological and phenotypical features. Flow cytometry analysis demonstrated uptake of acetylated low density lipoproteins (Ac-LDL) and constitutive expression of SLA class I, CD29, CD31, CD41/61, CD80/86, CD46, SWC3, and LAMP-1 antigens by all analyzed lines and showed little differences to primary cells. The functional similarity between primary and immortalized endothelial cells was demonstrated in a cytotoxicity assay using a human natural killer cell line (NKL) as effector. The AOCs cell lines should be valuable tools for in vitro study of the human immune response against pig endothelial cells. In addition, they would be very useful to gain insight in the pathogenesis of some viral haemorrhagic diseases of pig such as African swine fever (ASF) or classical swine fever (CSF).     

Role of Some Agricultural and Industrial By-products in Modifying Soil Physical Properties

In a pot culture study, saw dust, vermiculite, lignite, magnesium oxide, wheat straw, paddy shell, ground nut shell and farm yard manure (FYM) were incorporated in to wet sandy loam and clay soils at the rate of 30 g/kg. Relative changes in soil structure and moisture retention were evaluated at the end of 160 day incubation period. All the amendments except vermiculite, ground nut shell and FYM in sandy loam soil and except vermiculite, lignite, magnesium oxide in clay soil significantly increased MWD and aggregates > 1 mm. Structural index was significantly improved by all the amendments except vermiculite and FYM in sandy loam and vermiculite and magnesium oxide in clay soil. Moisture retention and release in clay soil in the wet range (0 to 0.33 bar) were considerably increased by soil amendments except magnesium oxide, vermiculite and lignite. Almost all the amendments significantly increased moisture retention in the wet range in sandy loam soil. However, moisture release at 50 mbar was reduced only by wheat straw in sandy loam soil.     

Busulphan and thiourea as chemosterilants for the snail lymnaea acuminata

Treatment of the snail Lymnaea acuminata with an alkylating agent (busulphan) or a thiocarbamide (thiourea), for 3 days, significantly reduced the number of eggs and prevented embryonic development. Both the drugs caused a dose dependent reduction in the levels of protein, DNA, RNA and the activity of the alkaline phosphatase of the ovotestis, and a dose-dependent enhancement in the levels of amino-acids and the activity of the acid phosphatase. Discontinuation of the drug treatment for 7 days resulted in a near complete recovery in DNA and RNA levels, while there was no change in the levels of protein, amino-acids and the activities of the acid and alkaline phosphatases.     

Solution of the kinematic-wave equations for border irrigation

A method is developed to obtain solutions to the kinematic-wave equations for border irrigation. The accuracy of this method is assessed by comparing the results with the kinematic-wave train (KWT) method previously reported to be accurate. Experimental data on freely draining borders were utilized for comparison and evaluation of these methods. The prediction error for advance time resulting from the two methods was comparable: it was below 25% for three sets of data but as high as 50% for one set of data. The prediction error for recession time was also comparable: below 6% for the same three sets of data but 14.0% for one set.     

Growth strategy of trees related to successional status II. Leaf dynamics

The leaf dynamics of two early (Alnus nepalensis and Schima khasiana) and three late (Machilus kingii, Quercus dealbata and Q. griffithii) successional tree species of a subtropical humid, evergreen, montane forest at Upper Shillong (1900 m) in north-eastern India has been studied using demographic techniques. The open-grown early successional species show greater leaf production and higher turnover rate than the open-grown late successional species. The latter, under forest situations, have still slower turnover rates but with leaves of several age groups. It is suggested that the early successional species are attuned to environments of temporary resource availability while the late successional species are adapted to the relatively predictable but highly competitive environments of closed forests.