Proximate Constituents Research Articles

Chemical and Physio-Chemical Characterization of the Flours and Oils from Whole and Rejected Cashew Nuts Cultivated in Southwest Nigeria

2 Abstract: Processed whole as well as rejected cashew nuts were obtained from a processing plant located at Owo, Ondo State, Nigeria. Each of them were divided into two, a part defatted while the other part were undefatted. They were thereafter characterized with respect to their proximate constituents, energy, mineral contents, anti-nutritional factors and functional properties. The physio-chemical properties of the oil from the cashew nuts were also determined. On the average, crude protein (CP) of the whole nut ranged from 200.0 ± 0.3 g kg DM in undefatted to 304.0 ± 0.1 g kg DM in the defatted sample while it ranged from 176.0 ± 0.6 -1 -1 g kg DM to 323.0 ± 0.5 g kg DM with 30% and 45.5% increments, respectively. The crude fat of th e -1 -1 kg . The water absorption capacity (WAC) was enhanced by 35% and 50%, respectively in the whole and -1 rejected samples by defatting while oil absorption capacity (OAC) were enhanced by 18% and 42%, respectively by defatting. There were wide variations among the foaming capacity (FC) and foaming stability (FS) as evident in the high values of coefficient of variation (CV) of 127.7% and 115.5% respectively. The protein solubility curves generally had multiple maxima and minima peaks. The oil from the CNM had iodine value of 115.0 wijs in rejects and 136.9 wijs in whole nut and the peroxide value varied widely with a CV of 41.85% when compared with other properties of the oil analyzed. The analytical data indicate that the good and rejected cashew nut could be important alternative protein and energy contributors to compound non ruminant animal feed in this region.

Glyphosate-Tolerant Alfalfa Is Compositionally Equivalent to Conventional Alfalfa (Medicago sativa L.)

Glyphosate-tolerant alfalfa (GTA) was developed to withstand over-the-top applications of glyphosate, the active ingredient in Roundup agricultural herbicides. As a part of the safety assessment, GTA (designated J101 x J163) was grown under controlled field conditions at geographically diverse locations within the United States during the 2001 and 2003 field seasons along with control and other conventional alfalfa varieties for compositional assessment. Field trials were conducted using a randomized complete block design with four replication blocks at each site. Alfalfa forage was harvested at the late bud to early bloom stage from each plot at five field sites in 2001 (establishment year) and from four field sites in 2003 (third year of stand). The concentration of proximate constituents, fibers, amino acids, coumestrol, and minerals in the forage was measured. The results showed that the forage from GTA J101 x J163 is compositionally equivalent to forage from the control and conventional alfalfa varieties.

Autotomy reduces feeding, energy storage and growth of the sea star Stichaster striatus

We evaluated the effect of autotomy on feeding, energy storage and growth of juvenile Stichaster striatus kept in the laboratory for five months with a limited supply of the mussel Semimytilus algosus. Autotomy strongly decreased feeding, energy storage and growth. Intact juveniles showed a ∼ 3 fold higher feeding rate than autotomized individuals throughout the experiment. Intact juveniles also had a higher (∼ 5 fold) energy content per pyloric caeca in each arm. This was mainly due to higher lipid content, the main proximate constituent of pyloric caeca. Intact juveniles showed a greater growth rate and reached a greater size than autotomized individuals, more evident for underwater mass than radius length. The reduced capacity to feed reduced energy intake in autotomized individuals. However, low energy reserves along with low growth in autotomized sea stars, support the hypothesis that juveniles of this species allocate energy to regeneration to the detriment of growth. This was also supported by the ∼ 25% of arm length regeneration after 5 mo. Remaining small could increase risk of lethal predation, however, S. striatus may reduce predation risk by using crevices and kelp holdfasts as refuges from predators. Given the strong impact of autotomy on feeding, regeneration of arms to recover full capacity to forage and grow seems a better strategy for juvenile S. striatus, than merely growing.

Exploration drilling optimisation using geostatistics: a case in Jharia Coalfield, India

Drill hole optimisation using geostatistics has been used in many parts of the world in respect of various mineral deposits including coal. The present paper is an attempt to provide a means to maximise the information to cost ratio for optimisation of exploration drilling in the Jharia coalfield, which is the only source of prime coking coal in India. Exploration in Jharia Coalfield that has led to the generation of a huge exploration database has been carried out by various governmental and non-governmental agencies. But because of lack of importance and an obvious application towards exploration modelling, the database had not been utilised to its fullest extent. In the present study, an attempt has been made to derive geostatistical models of various proximate coal constituents of selected seams in respect of nine opencast mine blocks of the coalfield. This experimental study, employing geostatistical volume–variance relationships, revealed that an exploration grid density of 16 drill holes per square kilometre with a grid spacing of 300 × 300 m within an exploration area of 1 × 1 km is adequate for maximisation of information in respect of the Jharia Coalfield. Until now, the density of exploration drilling in the coalfield has been considered as per Indian Standard Procedure (ISP) norms solely based on past experience and proposed mining methods. The present study of exploration drill hole optimisation using the concept of kriging variance is associated with an objectively defined mathematical basis and therefore may be applied for exploration optimisation in other coalfields in India and other countries, which occur in similar geological settings.

Nutritional traits of dorsal and ventral fillets from three farmed fish species

Dorsal and ventral fillet portions (DP and VP, respectively) of European sea bass ( Dicentrarchus labrax, ESB), gilthead sea bream ( Sparus aurata, GSB), and rainbow trout ( Oncorhynchus mykiss, RBT) were analysed for proximate constituents, and fatty acid composition and content. Moisture and lipid content differentiated DP from VP in all species. Significant differences emerged between DP and VP from ESB for MUFA, PUFA, and DHA contents. The n6/ n3 ratio ranged from 0.22 (DP in RBT) to 0.38 (VP in GSB). The highest hypocholesterolaemic/hypercholesterolaemic fatty acid ratio pertained to DP and VP from RBT, and the lowest peroxidisability index to VP from GSB and ESB. The index of nutritional quality for EPA + DHA was always higher in VP than in DP, that of RBT being especially interesting because it is associated with a lower energy value than that from ESB and GSB.

Indicators of Energetic Status in Juvenile Coho Salmon and Chinook Salmon

AbstractBioenergetic models frequently rely on published values or models for estimating the energy density of fish, principally because of the cost and effort of obtaining direct measurements. In this study, we developed empirical models of energy density for free‐ranging juvenile coho salmon Oncorhynchus kisutch and Chinook salmon O. tshawytscha sampled at sea from the west coast of Oregon to Kodiak Island, Alaska, and we evaluated the accuracy of published energy density models commonly used for these species. Our analyses showed that the energy density of juvenile coho and Chinook salmon was strongly correlated to percent dry weight and proximate constituents (especially lipid and, to a lesser extent, protein concentrations) but poorly correlated to body size and condition factor. Percent dry weight of whole fish was the single best predictor of energy density for both species, accounting for more than 90% of the variance in energy density. We also found that percent dry weight in the muscle tissue accounted for 65% of the variance in energy density. Changes in energy density mainly reflected changes in lipid composition. These results indicate that accurate estimates of energy density could be obtained at low effort and cost for juvenile coho and Chinook salmon simply by determining the water contents in whole‐fish or muscle samples. Published models overestimate the energy density of juvenile coho and Chinook salmon collected from the Pacific Ocean. This may result from the extrapolation of the models to different size‐classes, life stages, or habitats. More caution is needed when models are extrapolated to conditions beyond those that were used for their development.

Effects of Stage of Maturity and Cooking on the Chemical Composition of Select Mushroom Varieties

Select mushrooms were analyzed for proximate constituents and carbohydrate profiles either raw or cooked and at different stages of maturity. White button mushrooms (Agaricus biporus) contained high concentrations of ash (12.5 and 11.9% for immature and mature mushrooms, respectively). Starch and total dietary fiber (TDF) concentrations were higher in maitake (Grifola frondosa) and shiitake (Lentinus edodes) mushrooms. Crude protein (CP) and acid-hydrolyzed fat (AHF) were highest in crimini (Agaricus bisporus) and white button and maitake mushrooms, respectively. Chitin concentrations were highest in portabella (Agaricus bisporus) and enoki (Flammulina velutipes) mushrooms (8.0 and 7.7%, respectively). Oligosaccharides were found in low concentrations in some mushrooms. CP and TDF accounted for 86.4 and 49.3% of mushroom organic matter. Cooking increased starch, TDF, and AHF but decreased CP and chitin concentrations. The chitin concentration increased with mushroom maturity. These results detail the complete carbohydrate profile of several important mushroom varieties.

Utilization of lipids during early development of the sea urchin Evechinus chloroticus

Detailed studies of lipid utilization during bivalve development have shown that lipids are important at 2 critical periods: embryogenesis and metamorphosis. Using the latroscan TLC/FID system I examined lipid class utilization during development of Evechinus chloroticus to determine whether lipids were also important for an echinoderm at these times. Eggs of E. chloroticus contained 34.41 ng of lipid, primarily polar lipids (52.0 %) and triglyceride (29.4 %). To determine whether there was a different pattern of lipid utilization between larvae reared in the presence or absence of particulate food, larvae were either fed 6000 cells ml -1 Dunaliella every 2 d or starved. While there was no change in the amount of total, structural or energy storage lipids over time, there was a significant difference in the amount of structural lipids between Fed and Starved treatments. This was related to the continuing development of Fed larvae and cessation of development of Starved larvae at the 4-arm pluteus stage. In both treatments, triglycerides were rapidly utilized from the early 4-arm echinopluteus to the late 4-arm larva with fully developed arms and gut. Another neutral lipid, free fatty acid, accumulated in the 8-arm echinopluteus stage of the Fed larvae. This suggests that lipid stored during the planktonic phase, in combination with the proximate constituents of the larval body, provides the energy for the metamorphic and perimetamorphic periods in echinoderm development. Thus sea urchins, as in bivalves, appear to have 2 critical periods for lipid use during development.

Classification and prediction of maize hardness-associated properties using multivariate statistical analyses

Maize kernel hardness and proximate constituents were evaluated by several methods. Proximate constituents and hardness-associated properties were significantly correlated, and their correlation coefficients were improved when moisture, protein, and oil contents were kept constant. Multivariate techniques were applied to create new sets of variables to characterize maize hardness. The principal component scores created by principal component analysis were subjected to cluster analysis and discriminant analysis. Three factors interpreted as representing physical and chemical properties were identified using factor analysis. A total of 248 maize samples were grouped into 7 and 10 subgroups by cluster analysis. Beale's pseudo F statistic and the plot of principal component scores showed that 10 clusters would be a better solution than 7 clusters. The groups resulting from cluster analysis seem to have unique physical and chemical properties showing the different order in values of hardness measurements. Thus, the grouping by cluster analysis according to hardness-associated physical and chemical properties improved the explanation of maize kernel hardness. The application of discriminant analysis to create a classification rule for hardness clusters revealed that new observations had an 87% correct classification into hardness clusters.

Water Consumption of Yankasa Rams Fed a Basal Diet of Maize Stover-lablab Mixture

Sixteen Yankasa rams aged 18-24 months, weighing 25.4-31.0 kg were randomly divided into 4 treatment groups labelled A, B, C and D, each treatment comprising of 4 rams. Maize stover-lablab hay mixture in a 2:1 maize stover; lablab ratio was offered at 2, 4, 6 and 8% as body weight to treatments A, B, C and D respectively, in addition to 300g of concentrate per head per day in an individual feeding trial. Water was offered at 4kg per ram daily. Water refusal and left-over feed were recorded separately daily, before feeding in the morning. Samples of feed and faeces were analyzed separately for proximate constituents. Live weight changes were measured and used to adjust the levels of the basal diet throughout the 90 days duration of the trial. Results showed that the rams offered the higher level of the feed consumed more water than the rams on the other treatments. However, there was no significant difference (P>0.05) in wate r consumption between treatments D and C, and between treatments B and A. But treatment D wa s significantly different (P<0.05) from treatments B and A. Daily water consumption was 2.65, 2.63, 1.87 and 1.75 kg per ram for treatments D, C, B and A respectively. Yankasa rams offered high levels of maize stover- lablab hay mixture required about 3 litres of water per head day for optimum utilization of the feed.

Carbohydrate composition of selected plum/prune preparations.

Eighteen plum/prune preparations and byproducts were analyzed for proximate constituents and carbohydrate profiles. Plum puree and prune juice contained the highest concentrations of ash (13.0 and 13.8%, respectively). Crude protein (CP), acid-hydrolyzed fat (AHF), and total dietary fiber (TDF) concentrations were higher in byproducts (waste cake and dried plum pits) compared with the other fractions. Several classes of oligosaccharides were found in low concentrations in many of the substrates and were associated with the fruit rather than the pit. Maltooligosaccharides were found in very high concentrations in three of the preparations as a result of the addition of maltodextrin during processing. Monosaccharides, sugar alcohols, and oligosaccharides were found in higher concentrations in the fruit than in the pit and accounted for 2.9-84.7% of substrate organic matter. These results indicate that carbohydrates of various types constitute a significant proportion of plum/prune preparations and byproducts.

Proximate, vitamins A and E, and mineral composition of free‐ranging cotton mice (Peromyscus gossypinus) from St. Catherines Island, Georgia

AbstractAlthough rodents are an integral part of numerous carnivore diets, there is little published information regarding nutrient composition in free‐ranging mice for comparison with laboratory‐reared prey. Cotton mice (Peromyscus gossypinus, n=6) were captured on St. Catherines Island, Georgia, and analyzed for water, ash, protein, and fat content (proximate constituents), as well as minerals and the fat‐soluble vitamins A and E. The overall body composition (mean±SD: 65.8%±1.9% water, 10.9%±2.2% ash, 56.4%±4.1% protein, and 27.2%±3.9% fat) was similar to published values for adult laboratory mice (Mus domesticus). The macro‐ (Ca, K, Mg, Na, and P) and trace (Cu, Fe, Mn, and Zn) mineral levels were also similar to previously reported values for laboratory mice, and in general met or exceeded the established nutrient requirements for domestic carnivores. Vitamin E ranged from 77 to 170 IU/kg dry matter (DM) in these mice–again, similar to values previously quantified in laboratory mice. However, the vitamin A concentrations (21,947±6,893 IU/kg DM) in the free‐ranging mice were consistently and substantially lower than values reported for whole laboratory mice. To our knowledge, this is the first quantification of vitamin A in free‐ranging mice used as prey by carnivores. While other nutrients measured were similar between captive‐reared and free‐ranging mice, the current data suggest the need for further investigation of vitamin A nutrition in the development of optimal diets for carnivores in captivity. Zoo Biol 23:253–261, 2004. © 2004 Wiley‐Liss, Inc.

Nutritional Status of the Protein of Corn-Soy Based Extruded Products Evaluated by Rat Bioassay

A rat bioassay was conducted to preclinically evaluate nutritional quality of two supplementary foods (SFs) developed based on corn and soy blends for feeding preschool children. The SFs prepared by extrusion cooking and subsequently modified to taste either sweet or salty provide 395 +/- 2 kcal of energy and 20 +/- 2 g protein per 100 g of food. The proximate constituents and energy contents of SFs were within the ranges prescribed for processed weaning foods and could satisfactorily meet the requirements of preschool children. Groups of male weanling rats were fed SFs for 4 weeks to evaluate the protein quality. The body weight gain of rats fed with SFs were significantly higher than those fed with skimmed milk powder (SMP) diet as control. The protein efficiency ratio and net protein utilization results of SFs were not significantly different (p > 0.05) from values of control group. It is inferred that these SFs were nutritionally comparable to SMP.

Exploiting dietary fibre in equid diets

AbstractFour in vivo apparent digestibility trials were carried out over a period of two years using 4 to 6 Welsh cross pony geldings of approximately 300kg LW. In all experiments ponies were individually housed in 10x12 ft loose–boxes with rubber mat flooring. In Experiment 1 three diets were offeredad libitumto 6 ponies in a replicate 3 x 3 Latin squares design. In Experiments 2, 3 &amp; 4, four ponies were offered 1.5 – 1.75 % of LW per day in dry matter (DM) in 4 x 4 Latin square changeover designs. In all experiments periods lasted for 21 days, a 16-day adaptation period followed by a 5-day data collection phase. Diets were fed in two equal meals per day and water was availablead libitum. Dried feed and faecal samples were analysed for DM, ADF, NDF and CP using the methods of the AOAC (1990), GE was determined via bomb calorimetry and non–starch polysaccharides (NSP) were quantified by the method of Englyst and Cummings (1984). Differences between diets were determined by analysis of variance (Genstat 5, 2000). NSP analysis gave detailed information on the individual fibre constituents and allowed the subtle differences between feeds to be identified. The DMI of grass and red clover silages were 0.6 and 1.7 kg/d greater than hay indicating that the low DM content of these two forages does not negatively affect feed intake. The apparent digestibilities of these two feeds were also significantly greater than hay for all parameters measured. In Experiment 2 the combination diets of alfa-beet and hay (AB:H) and sugar beet and hay (SB:H) fed in 60:40 and 70:30 ratios were all significantly more digested at &gt; 50% than the hay only diet at &lt;39% for DM, CP, NDF, ADF, NSP, Ca, P and Mg. The addition of AB to H in the 70:30 diet produced a DE value of 2.89 MJ/kg DM higher than the hay alone only diet and also caused an associative affect which improved the digestibility of CP, arabinose, galactose and uronic acids from the hay portion of the diet. No significant differences were noted between the digestibilities of the proximate constituents for the chopped hay replacer (CHR), CHR and highly digestible fibre (HDF) cubes and CHR and course mix (CM) fed in Experiment 3. When looking at the digestibilities of the individual NSP constituents the CHR : CM diet was generally less digestible than the CHR: HDF diets indicating that HDF cubes are equally well or more digestible than a diet containing conventional course mix. With DE values similar to the CM the HDF feed can be offered as a valuable alternative to concentrates in equid diets. The high fibre forage (HiFi) fed in Experiment 4 was also highly digestible for all constituents measured and showed that a combination of chopped straw and alfalfa makes an excellent fibre feed for horses. The results from the four experiments reported here show that detailed chemical analysis is extremely valuable when choosing complimentary fibre feeds for horses. Moreover all the fibre feeds examined here were more digestible than hay. Feeds like the HDF cubes can be used to replace some or all of the cereal portion of the diet. The associative effects between the alfa-beet and hay and the mineral digestibility from these alternative feeds also suggests that not only is gut health enhanced, but that the supply of nutrients from these feeds makes them ideal choices for stabled horses.

Quality of farmed salmonids with emphasis on proximate composition, yield and sensory characteristics

Salmonid aquaculture has focused for many years on enhancing the quantity of fish produced. However, optimization of the quality of salmonids may lead to improved consumer acceptance and higher prices for the farmed product. The present review evaluates how the quality of salmonids is affected by parameters such as feed type, level of dietary intake (ration) and growth. Feed composition has a major influence on the proximate composition of salmonids. In particular, whole body lipid as well as the lipid content in the edible fillet is directly related to dietary fat content, while the fatty acid composition of the fish flesh is also strongly influenced by the dietary fatty acid profile. While fish body composition appears to be largely influenced by feed composition, an increase in other parameters such as feed ration and fish size also results in enhanced adipose deposition and decreased water content in the fish body. The protein content, however, remains more or less stable. An increase in fish body lipid content is not necessarily a negative factor, depending upon the processing procedure that follows. However, an increase in body fat content is generally accompanied by a reduction in slaughter yield, owing to an increase in the weight of viscera in relation to body weight. Although salmonid flesh quality with respect to levels of proximate constituents and yield appears to be under the strong influence of feed composition and feed amount, the sensory characteristics, such as odour, flavour, texture, etc., are only governed by these factors to a small degree. Tactics for the rearing of salmonids for specific purposes should therefore take into consideration the fact that the level of proximate constituents in the whole body as well as the fillet are readily manipulated by feed composition and feeding strategies, whereas the sensory parameters are less affected by these variables.

In vivo apparent digestibility of proximate constituents and non-starch polysaccharides in ponies offered botanically diverse fibre based diets

Little information is available on digestion of the non-starch polysaccharide (NSP) fraction of fibrous feeds in equines. Two studies were conducted which examined the in vivo apparent digestibilities of proximate constituents and NSP in ponies offered diets based on botanically diverse fibrous foodstuffs.In study 1 (S1), three mature caecally-fistulated Welsh-cross pony geldings (266 kg LW) were used in a 3 x 3 latin square changeover design experiment consisting of three 21 day periods. Ponies were offered 4 kg dry matter (DM) per day of either unmolassed sugar beet pulp (USBP), hay cubes (HC) or a 2:1 mix of oat hulls:naked oats (OHNO) plus minerals in 2 equal meals per day. After completion of S1 the same 3 ponies were used in study 2 (S2) where they were offered 4 kg DM/day of a 50:50 mix of USBP:HC (USHC) plus minerals fed as in S1 for a 21 day period.

Studies on deep-fat fried snacks from some cereals and legumes

Deep-fat fried products, prepared from cereals, legumes and their blends, are universally popular due to their desirable organoleptic profile. Grain types differ in their chemical make-up and physicochemical properties, which may account for the differences in the oil content of the snacks. In addition, the frying medium also plays an important role in the oil pick-up during deep-fat frying. This work attempts to correlate these parameters with the oil content in a traditional Indian extruded and deep-fat fried product, sev, using flours from rice, wheat, amaranth, chickpea, cowpea, black gram and green gram. No correlation between the proximate constituents (water-holding capacity and oil-holding capacity) and the oil content of the fried sev was seen. Oil content in sev was minimum when fried in cottonseed oil. © 1998 SCI.

Nutrient Composition of Whole Green Frogs, Rana clamitans and Southern Toads, Bufo terrestris

ABSTRACT Whole prey samples of green frogs, Rana clamitans, and southern toads, Bufo terrestris, were analyzed for proximate composition (moisture, lipid, protein, ash), fat-soluble vitamins A and E, and selected minerals. The green frogs (n=7) were obtained from a pond in Cold Springs Harbor, NY in August 1996. The southern toads (n=5) were collected in July 1996 on St. Catherine's Island, GA. Water (77.5 ± 1.3 vs. 71.8 ± 2.9, %) and crude protein (71.2 ± 3.3 vs. 61.0 ± 1.2, % DM) percentages were significantly higher in frogs than in toads; all other proximate constituents were similar. Vitamin A was extremely variable, but averaged approximately 29,000 IU/kg DM (both species), while vitamin E in frogs was significantly lower (82.2 ± 1 5 .6 IU/kg DM) than in toads (369.3 ± 169.0 IU/kg DM). Frogs contained extremely high magnesium levels (2.5 ± 0.1 %DM) that may be considered excessive and affect mineral nutrition. Frogs also contained significantly higher values of calcium and sodium than toads. Copper,...

The Use of the InfraAlyzer 260 Whole Grain for Water and Fat Determination in Pork Meat

This study describes an investigation into the utility of the filter near infarared (NIR) instrument – InfraAlyzer 260 Whole Grain (Bran+Luebbe GmbH) for the routine analysis of pork meat composition in a rapid and cost-effective manner. Water and fat were the proximate constituents selected for investigation on the grounds of their importance in pork meat processing and marketing. Fresh slices of Longissimus dorsi (LD) muscle weighing approximately 300 g were taken from carcasses. The LD samples were then cut and homogenized with a rotating knife homogenizer prior to NIR and chemical analysis according to the ISO-procedures. The selected samples ( n = 79) were scanned in an open sample cup using all 19 filters of an InfraAlyzer 260 Whole Grain with wavelenghts between 1110 and 2310 nm. The process of calibration (MLR method) was performed by using SESAME ver. 2.10 software (Bran+Luebbe GmbH) on an interfaced PC computer. Standard errors of calibration were 0.16 % for water and 0.17 % for fat. Calibrations were validated with an independent set of 21 samples of the same types. Standard errors of validation were 0.17 % and 0.20 % for water and fat, respectively. The obtained results show that InfraAlyzer 260 Whole Grain can be used for the rapid and accurate analysis of components of the pork meat.

Use of cane condensed molasses solubles in feeding broilers

The mean composition of condensed molasses solubles (vinasse or stillage) obtained from the three alcohol fermentation plants of the sugar cane industry including proximate constituents, inorganic components and amino acid composition was obtained. Mean solids content was 620 g kg −1. Vinasse found to be low in fat and fiber but high in ash and different carbohydrate contents. Potassium content of ash was almost 500 g kg −1 which may be more a disadvantage than an advantage in poultry feeding. Ash can contribute substantially to the supply of calcium, magnesium, sodium, iron and chloride in poultry feed. Vinasse was low in protein and the total content of essential amino acids including lysine and methionine was average. Performance results of broilers fed up to 60 g kg −1 of neutralized stillage have shown that low levels of up to 40 g kg −1 can prove profitable for the broiler feed industry, chiefly in sugar cane producing countries where its price is low and it is considered a waste by-product that may pollute.