International Journal of Biological, Life and Agricultural Sciences

Predicting Long-Term Meat Productivity for the Kingdom of Saudi Arabia

Year: 2015 Volume: 9 Issue: 3 304 - 308 Pages

Abstract: Livestock is one of the fastest-growing sectors in agriculture. If carefully managed, have potential opportunities for economic growth, food sovereignty and food security. In this study we mainly analyse and compare long-term i.e. for year 2030 climate variability impact on predicted productivity of meat i.e. beef, mutton and poultry for the Kingdom of Saudi Arabia w.r.t three factors i.e. i) climatic-change vulnerability ii) CO2 fertilization and iii) water scarcity and compare the results with two countries of the region i.e. Iraq and Yemen. We do the analysis using data from diverse sources, which was extracted, transformed and integrated before usage. The collective impact of the three factors had an overall negative effect on the production of meat for all the three countries, with adverse impact on Iraq. High similarity was found between CO2 fertilization (effecting animal fodder) and water scarcity i.e. higher than that between production of beef and mutton for the three countries considered. Overall, the three factors do not seem to be favorable for the three Middle-East countries considered. This points to possibility of a vegetarian year 2030 based on dependency on indigenous livestock population.

Effect of Three Sand Types on Potato Vegetative Growth and Yield

Year: 2015 Volume: 9 Issue: 3 300 - 303 Pages

Abstract: Potato (Solanum tuberosum L.) is one of the major vegetable crops that are grown world –wide because of its economic importance. This experiment investigated the effect of local sands (River Base, Al-Ekader and Karbala) on number and total weight of minitubers. Statistical analysis revealed that there were no significant differences among sand cultures in number of stem/plant, chlorophyll index and tubers dry weight. River Base sand had the highest plant height (74.9 cm), leaf number/plant number (39.3), leaf area (84.4 dcm2⁄plant), dry weight/plant (26.31), tubers number/plant (8.5), tubers weight/plant (635.53 gm) and potato tuber yields/trove (28.60 kg), whereas the Karbala sand had lower performance. All the characters had positive and significant correlation with yields except the traits number of stem and tuber dry weight.

Influence of Cyperus rotundus Active Principles Inhibit Viral Multiplication and Stimulate Immune System in Indian White Shrimp Fenneropenaeus indicus against White Spot Syndrome Virus Infection

Year: 2015 Volume: 9 Issue: 4 360 - 364 Pages

Abstract: The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, FIII to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological, and immunological parameters. Surprisingly, the pooled fractions (F-IV to FVI) incorporated diets helped to significantly (P

Removal of Polycyclic Aromatic Hydrocarbons Present in Tyre Pyrolytic Oil Using Low Cost Natural Adsorbents

Year: 2015 Volume: 9 Issue: 2 186 - 190 Pages

Authors:
Neha Budhwani

Abstract: Polycyclic aromatic hydrocarbons (PAHs) are formed during the pyrolysis of scrap tyres to produce tyre pyrolytic oil (TPO). Due to carcinogenic, mutagenic, and toxic properties PAHs are priority pollutants. Hence it is essential to remove PAHs from TPO before utilising TPO as a petroleum fuel alternative (to run the engine). Agricultural wastes have promising future to be utilized as biosorbent due to their cost effectiveness, abundant availability, high biosorption capacity and renewability. Various low cost adsorbents were prepared from natural sources. Uptake of PAHs present in tyre pyrolytic oil was investigated using various low-cost adsorbents of natural origin including sawdust (shisham), coconut fiber, neem bark, chitin, activated charcoal. Adsorption experiments of different PAHs viz. naphthalene, acenaphthalene, biphenyl and anthracene have been carried out at ambient temperature (25°C) and at pH 7. It was observed that for any given PAH, the adsorption capacity increases with the lignin content. Freundlich constant Kf and 1/n have been evaluated and it was found that the adsorption isotherms of PAHs were in agreement with a Freundlich model, while the uptake capacity of PAHs followed the order: activated charcoal> saw dust (shisham) > coconut fiber > chitin. The partition coefficients in acetone-water, and the adsorption constants at equilibrium, could be linearly correlated with octanol–water partition coefficients. It is observed that natural adsorbents are good alternative for PAHs removal. Sawdust of Dalbergia sissoo, a by-product of sawmills was found to be a promising adsorbent for the removal of PAHs present in TPO. It is observed that adsorbents studied were comparable to those of some conventional adsorbents.

Evaluation of Bioactive Phenols in Blueberries from Different Cultivars

Year: 2015 Volume: 9 Issue: 4 356 - 359 Pages

Abstract: Blueberries are widely valued for their high content in phenolic compounds with antioxidant activity, and hence beneficial for the human health. In this way, a study was done to determine the phenolic composition (total phenols, anthocyanins and tannins) and antioxidant activity of blueberries from three cultivars (Duke, Bluecrop, and Ozarkblue) grown in two different Portuguese farms. Initially two successive extractions were done with methanol followed by two extractions with aqueous acetone solutions. These extracts obtained were then used to evaluate the amount of phenolic compounds and the antioxidant activity. The total phenols were observed to vary from 4.9 to 8.2 mg GAE/g fresh weight, with anthocyanin’s contents in the range 1.5-2.8 mg EMv3G/g and tannins contents in the range 1.5- 3.8 mg/g. The results for antioxidant activity ranged from 9.3 to 23.2 molTE/g and from 24.7 to 53.4molTE/g, when measured, respectively, by DPPH and ABTS methods. In conclusion it was observed that, in general, the cultivar had a visible effect on the phenols present, and furthermore, the geographical origin showed relevance either in the phenols contents or the antioxidant activity.

Land Suitability Analysis for Maize Production in Egbeda Local Government Area of Oyo State Using GIS Techniques

Year: 2015 Volume: 9 Issue: 3 294 - 299 Pages

Abstract: Maize constitutes a major agrarian production for use by the vast population but despite its economic importance; it has not been produced to meet the economic needs of the country. Achieving optimum yield in maize can meaningfully be supported by land suitability analysis in order to guarantee self-sufficiency for future production optimization. This study examines land suitability for maize production through the analysis of the physicochemical variations in soil properties and other land attributes over space using a Geographic Information System (GIS) framework. Physicochemical parameters of importance selected include slope, landuse, physical and chemical properties of the soil, and climatic variables. Landsat imagery was used to categorize the landuse, Shuttle Radar Topographic Mapping (SRTM) generated the slope and soil samples were analyzed for its physical and chemical components. Suitability was categorized into highly, moderately and marginally suitable based on Food and Agricultural Organisation (FAO) classification, using the Analytical Hierarchy Process (AHP) technique of GIS. This result can be used by small scale farmers for efficient decision making in the allocation of land for maize production.

Effects of Process Parameters on the Yield of Oil from Coconut Fruit

Year: 2015 Volume: 9 Issue: 3 289 - 293 Pages

Abstract: Analysis of the properties of coconut (Cocos nucifera) and its oil was evaluated in this work using standard analytical techniques. The analyses carried out include proximate composition of the fruit, extraction of oil from the fruit using different process parameters and physicochemical analysis of the extracted oil. The results showed the percentage (%) moisture, crude lipid, crude protein, ash and carbohydrate content of the coconut as 7.59, 55.15, 5.65, 7.35 and 19.51 respectively. The oil from the coconut fruit was odourless and yellowish liquid at room temperature (30oC). The treatment combinations used (leaching time, leaching temperature and solute: solvent ratio) showed significant differences (P

The Relations of Volatile Compounds, Some Parameters and Consumer Preference of Commercial Fermented Milks in Thailand

Year: 2014 Volume: 8 Issue: 9 1069 - 1072 Pages

Abstract: The aim of research was to define the relations between volatile compounds, some parameters (pH, titratable acidity (TA), total soluble solid (TSS), lactic acid bacteria count) and consumer preference of commercial fermented milks. These relations tend to be used for controlling and developing new fermented milk product. Three leading commercial brands of fermented milks in Thailand were evaluated by consumers (n=71) using hedonic scale for four attributes (sweetness, sourness, flavour, and overall liking), volatile compounds using headspace-solid phase microextraction (HS-SPME) GC-MS, pH, TA, TSS and LAB count. Then the relations were analyzed by principal component analysis (PCA). The PCA data showed that all of four attributes liking scores were related to each other. They were also related to TA, TSS and volatile compounds. The related volatile compounds were mainly on fermented produced compounds including acetic acid, furanmethanol, furfural, octanoic acid and the volatiles known as artificial fruit flavour (beta pinene, limonene, vanillin, and ethyl vanillin). These compounds were provided the information about flavour addition in commercial fermented milk in Thailand.

Effect of Germination on Proximate, Available Phenol and Flavonoid Content, and Antioxidant Activities of African Yam Bean (Sphenostylis stenocarpa)

Year: 2015 Volume: 9 Issue: 1 106 - 109 Pages

Abstract: The work studied the effect of germination on proximate, phenol and flavonoid content and antioxidant activities (AOA) of African Yam been (AYB). Germination was done in controlled dark chamber (100% RH, 28oC). The proximate, phenol and flavonoid content and antioxidant activities before and after germination were investigated. The crude protein, moisture, and crude fiber content of germinated AYB were significantly higher (P

Effectiveness of Biopesticide against Insects Pest and Its Quality of Pomelo (Citrus maxima Merr.)

Year: 2015 Volume: 9 Issue: 3 285 - 288 Pages

Abstract: Effect of biopesticide from wood vinegar and extracted substances from 3 medicinal plants such as: non taai yak (Stemona tuberosa Lour), boraphet (Tinospora crispa Mier) and derris (Derris elliptica Roxb) were tested on the age five years of pomelo. The selected pomelo was carried out for insects’ pest control and its quality. The experimental site was located at farmer’s orchard in Phichit Province, Thailand. This study was undertaken during the drought season (December to March). The extracted from plants and wood vinegar were evaluated in 6 treatments: 1) water as control; 2) wood vinegar; 3) S. tuberosa Lour; 4) T. crispa Mier; 5) D. elliptica Roxb; 6) mixed (wood vinegar + S. tuberosa Lour + T. crispa Mier + D. elliptica Roxb). The experiment was RCB with 6 treatments and 3 replications per treatment. The results showed that T. crispa Mier was the highest effectiveness for reduction population of thrips (Scirtothrips dorsalis Hood) and citrus leaf miner (Phyllocnistis citrella Stainton) at 14.10 and 15.37 respectively, followed by treatment of mixed, D. elliptica Roxb, S. tuberosa Lour and wood vinegar with significance different. Additionally, T. crispa Mier promoted the high quality of harvested pomelo in term of thickness of skin at 12.45 mm and S. tuberosa Lour gave the high quality of the pomelo in term of firmness (276.5 kg/cm2) and brix (11.0%).

Impact of Two Herbal Seeds Supplementation on Growth Performance and Some Biochemical Blood and Tissue Parameters of Broiler Chickens

Year: 2015 Volume: 9 Issue: 3 279 - 284 Pages

Abstract: The effects of basil and/or chamomile seed supplementation on the growth of Hubbard broiler chicks were evaluated. The antioxidant effects of these supplements were also assessed. 120 1-day-old broiler chicks were randomly divided into four equal groups. The control group (group 1) was fed a basal diet (BD) without supplementation. Groups 2, 3, and 4 were fed the BD supplemented with 10g basil, 10g chamomile, and 5g basil plus 5g chamomile per kg of food, respectively. Basil supplementation alone or in combination with chamomile non-significantly (P≥0.05) increased final body weight (3.2% and 0.3%, respectively) and weight gain (3.5% and 3.6%, respectively) over the experimental period. Chamomile supplementation alone non-significantly (P≥0.05) reduced final body weight and weight gain over the experimental period by 1.7% and 1.7%, respectively. In comparison to the control group, herbal seed supplementation reduced feed intake and improved the feed conversion and protein efficiency ratios. In general, basil seed supplementation stimulated chicken growth and improved the feed efficiency more effectively than chamomile seed supplementation. The antioxidant activities of basil and/or chamomile supplementation were examined in the thymus, bursa, and spleen. In chickens that received supplements, the level of malondialdehyde was significantly decreased, whereas the activities of glutathione, superoxide dismutase, and catalase were significantly increased (P

Sound Exposure Effects towards Ross Broilers Growth Rate

Year: 2015 Volume: 9 Issue: 1 101 - 105 Pages

Abstract: Sound exposure effects have been investigated by broadcasting a group of broilers with sound of Quran verses (Group B) whereas the other group is the control broilers (Group C). The growth rate comparisons in terms of weight and raw meat texture measured by shear force have been investigated. Twenty-seven broilers were randomly selected from each group on Day 24 and weight measurement was carried out every week till the harvest day (Day 39).Group B showed a higher mean weight on Day 24 (1.441 ± 0.013 kg) than Group C. Significant difference in the weight on Day 39 existed for Group B compared to Group C (p < 0.05). However, there was no significant (p >0.05) difference of shear force in the same muscles (breast and drumstick raw meat) of both groups but the shear force of the breast meat for Group B and C broilers was lower (p < 0.05) than that of their drumstick meat. Thus, broadcasting the sound of Quran verses in the coop can be applied to improve the growth rate of broilers for producing better quality poultry.

Antimicrobial Properties of Copper in Gram-Negative and Gram-Positive Bacteria

Year: 2015 Volume: 9 Issue: 3 274 - 278 Pages

Abstract: For centuries humans have used the antimicrobial properties of copper to their advantage. Yet, after all these years the underlying mechanisms of copper mediated cell death in various microbes remain unclear. We had explored the hypothesis that copper mediated increased levels of lipid peroxidation in the membrane fatty acids is responsible for increased killing in Escherichia coli. In this study we show that in both gram positive (Staphylococcus aureus) and gram negative (Pseudomonas aeruginosa) bacteria there is a strong correlation between copper mediated cell death and increased levels of lipid peroxidation. Interestingly, the non-spore forming gram positive bacteria as well as gram negative bacteria show similar patterns of cell death, increased levels of lipid peroxidation, as well as genomic DNA degradation, however there is some difference in loss in membrane integrity upon exposure to copper alloy surface.

Bifidobacterium lactis Fermented Milk Was Not Effective for Helicobacter pylori Eradication: A Prospective, Randomized, Double-Blind, Controlled Study

Year: 2015 Volume: 9 Issue: 3 270 - 273 Pages

Abstract: The management of Helicobacter pylori (H. pylori) eradication is still a matter of discussion, full effectiveness is rarely achieved, and it has many adverse effects. The use of probiotics may be associated with better eradication rates and possibly prevention of adverse events due to antibiotic therapy. The present clinical study was undertaken to evaluate the efficacy of a specially designed fermented milk product, containing Bifidobacterium lactis B420, on the eradication of H. pylori infection in a prospective, randomized, double-blind, controlled study in humans. Four test fermented milks (FM) were specially designed in which counts of viable cells in all products were 10^10 Log CFU. 100 mL-1 for Bifidobacterium lactis - Bifidobacterium species 420. 190 subjects infected with H. pylori, with previous diagnosis of functional dyspepsia according to Rome III criteria entered the study. Bifidobacterium lactis B420, administered twice a day for 90 days was not able to eradicate H. pylori in Brazilian patients with functional dyspepsia.

'Pink' Waxapple Response to Salinity: Growth and Nutrient Uptake

Year: 2015 Volume: 9 Issue: 3 266 - 269 Pages

Abstract: Waxapple (Syzygium samarngense Merr.et Perry) is an important tropical fruit in Taiwan. The famous producing area is located on the coast in Pingtung County. Land subsidence and climate change will tend to soil alkalization more seriously. This study was to evaluate the effects of NaCl in waxapple seedlings. NaCl salinity reduced waxapple shoot growth; it may due to reducing relative water content in leaf and new shoot. Leaf Cl and Na concentration were increased but K, Ca, and Mg content had no significant difference after irrigated with NaCl for six weeks. In roots, Na and Cl content increase significantly with 90 mM NaCl treatment, but K, Ca, and Mg content was reduced. 30-90mM Nacl treatment do not effect K/Na, Ca/Na and Mg/Na ratio, but decrease significantly in 90mM treatment in roots. The leaf and root electrolyte leakage were significantly affected by 90 mM NaCl treatment. Suggesting 90mM was optimum concentration for sieve out other tolerance waxapples verities.

Selection of Saccharomyces cerevisiae Strains Tolerant to Lead and Cadmium Toxicity

Year: 2015 Volume: 9 Issue: 1 93 - 100 Pages

Abstract: The aim of this study was to select the best strains of Saccharomyces cerevisiae able to resist lead and cadmium. Ten strains were screened on the basis of their resistance at different concentrations of 0, 2, 4, 8, 12, 16, 20 and 24 ppm for Pb and 0, 0.5, 1, 2, 4, 6, 8 and 10 ppm for Cd. The properties of baker's yeast quality were decreased by the increase of Pb or Cd in growth medium. The slope values of yield, total viable cells and gassing power of produced baker's yeast were investigated as an indicator of metal resistant. In addition, concentrations of Pb and Cd in produced baker's yeast were determined. The strain of S. cerevisiae FH-620 had the highest resistance against Pb and Cd and had the minimum levels of both two investigated metals in produced baker's yeast.

Detection of Transgenes in Cotton (Gossypium hirsutum L.) by Using Biotechnology/Molecular Biological Techniques

Year: 2015 Volume: 9 Issue: 1 87 - 92 Pages

Abstract: Agriculture is the backbone of economy of Pakistan and cotton is the major agricultural export and supreme source of raw fiber for our textile industry. To combat severe problems of insect and weed, combination of three genes namely Cry1Ac, Cry2A and EPSPS genes was transferred in locally cultivated cotton variety MNH-786 with the use of Agrobacterium mediated genetic transformation. The present study focused on the molecular screening of transgenic cotton plants at T3 generation in order to confirm integration and expression of all three genes (Cry1Ac, Cry2A and EPSP synthase) into the cotton genome. Initially, glyphosate spray assay was used for screening of transgenic cotton plants containing EPSP synthase gene at T3 generation. Transgenic cotton plants which were healthy and showed no damage on leaves were selected after 07 days of spray. For molecular analysis of transgenic cotton plants in the laboratory, the genomic DNA of these transgenic cotton plants were isolated and subjected to amplification of the three genes. Thus, seventeen out of twenty (Cry1Ac gene), ten out of twenty (Cry2A gene) and all twenty (EPSP synthase gene) were produced positive amplification. On the base of PCR amplification, ten transgenic plant samples were subjected to protein expression analysis through ELISA. The results showed that eight out of ten plants were actively expressing the three transgenes. Real-time PCR was also done to quantify the mRNA expression levels of Cry1Ac and EPSP synthase gene. Finally, eight plants were confirmed for the presence and active expression of all three genes at T3 generation.

Application of Statistical Approach for Optimizing CMCase Production by Bacillus tequilensis S28 Strain via Submerged Fermentation Using Wheat Bran as Carbon Source

Year: 2015 Volume: 9 Issue: 1 76 - 86 Pages

Abstract: Biofuels production has come forth as a future technology to combat the problem of depleting fossil fuels. Bio-based ethanol production from enzymatic lignocellulosic biomass degradation serves an efficient method and catching the eye of scientific community. High cost of the enzyme is the major obstacle in preventing the commercialization of this process. Thus main objective of the present study was to optimize composition of medium components for enhancing cellulase production by newly isolated strain of Bacillus tequilensis. Nineteen factors were taken into account using statistical Plackett-Burman Design. The significant variables influencing the cellulose production were further employed in statistical Response Surface Methodology using Central Composite Design for maximizing cellulase production. The optimum medium composition for cellulase production was: peptone (4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L), Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride (0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at 37oC. Analysis of variance (ANOVA) revealed high coefficient of determination (R2) of 0.99. Maximum cellulase productivity of 11.5 IU/ml was observed against the model predicted value of 13 IU/ml. This was found to be optimally active at 60oC and pH 5.5.

Management of Meskit (Prosopis juliflora) Tree in Oman: The Case of Using Meskit (Prosopis juliflora) Pods for Feeding Omani Sheep

Year: 2015 Volume: 9 Issue: 1 70 - 75 Pages

Abstract: This study evaluated the use of raw or processed Prosopis juliflora (Meskit) pods as a major ingredient in a formulated ration to provide an alternative non-conventional concentrate for livestock feeding in Oman. Dry Meskit pods were reduced to lengths of 0.5- 1.0 cm to ensure thorough mixing into three diets. Meskit pods were subjected to two types of treatments; roasting and soaking. They were roasted at 150оC for 30 minutes using a locally-made roasting device (40 kg barrel container rotated by electric motor and heated by flame gas cooker). Chopped pods were soaked in tap water for 24 hours and dried for 2 days under the sun with frequent turning. The Meskit-pod-based diets (MPBD) were formulated and pelleted from 500 g/kg ground Meskit pods, 240 g/kg wheat bran, 200 g/kg barley grain, 50 g/kg local dried sardines and 10 g/kg of salt. Twenty four 10 months-old intact Omani male lambs with average body weight of 27.3 kg (± 0.5 kg) were used in a feeding trial for 84 days. They were divided (on body weight basis) and allocated to four diet combination groups. These were: Rhodes grass hay (RGH) plus a general ruminant concentrate (GRC); RGH plus raw Meskit pods (RMP) based concentrate; RGH plus roasted Meskit pods (ROMP) based concentrate; RGH plus soaked Meskit pods (SMP) based concentrate Daily feed intakes and bi-weekly body weights were recorded. MPBD had higher contents of crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) than the GRC. Animals fed various types of MPBD did not show signs of ill health. There was a significant effect of feeding ROMP on the performance of Omani sheep compared to RMP and SMP. The ROMP fed animals had similar performance to those fed the GRC in terms of feed intake, body weight gain and feed conversion ratio (FCR).This study indicated that roasted Meskit pods based diet may be used instead of the commercial concentrate for feeding Omani sheep without adverse effects on performance. It offers a cheap alternative source of protein and energy for feeding Omani sheep. Also, it might help in solving the spread impact of Meskit trees, maintain the ecosystem and helping in preserving the local tree species.

Gammarus:Asellus Ratio as an Index of Organic Pollution – (A Case Study in Markeaton, Kedleston Hall, and Allestree Park Lakes Derby) UK

Year: 2015 Volume: 9 Issue: 3 256 - 265 Pages

Authors:
U. Bawa

Abstract: Macro invertebrates have been used to monitor organic pollution in rivers and streams. Several biotic indices based on macro invertebrates have been developed over the years including the Biological Monitoring Working Party (BMWP). A new biotic index, the Gammarus:Asellus ratio has been recently proposed as an index of organic pollution. This study tested the validity of the Gammarus:Asellus ratio as an index of organic pollution, by examining the relationship between the Gammarus:Asellus ratio and physical chemical parameters, and other biotic indices such as BMWP and, Average Score Per Taxon (ASPT) from lakes and streams at Markeaton Park, Allestree Park and Kedleston Hall, Derbyshire. Macro invertebrates were sampled using the standard five minute kick sampling techniques physical and chemical environmental variables were obtained based on standard sampling techniques. Eighteen sites were sampled, six sites from Markeaton Park (three sites across the stream and three sites across the lake). Six sites each were also sampled from Allestree Park and Kedleston Hall lakes. The Gammarus:Asellus ratio showed an opposite significant positive correlations with parameters indicative of organic pollution such as the level of nitrates, phosphates, and calcium and also revealed a negatively significant correlations with other biotic indices (BMWP/ASPT). The BMWP score correlated positively significantly with some water quality parameters such as dissolved oxygen and flow rate, but revealed no correlations with other chemical environmental variables. The BMWP score was significantly higher in the stream than the lake in Markeaton Park, also The ASPT scores appear to be significantly higher in the upper Lakes than the middle and lower lakes. This study has further strengthened the use of BMWP/ASPT score as an index of organic pollution. But additional application is required to validate the use of Gammarus:Asellus as a rapid bio monitoring tool.