Inhibition of amylases from different origins by albumins from the wheat kernel.

The amylase activity of water extracts from 18 insect species, from 23 marine species and from 17 different species of birds and mammals was determined quantitatively. The inhibition of amylase in these extracts by three albumin fractions from the mature wheat kernel, which had been separated according to their molecular weights (60 000, 24 000 and 12 500 D), was determined as well. The inhibition activity of the three albumin fractions toward amylases extracted from a number of cereal species or from immature and germinating wheat kernel was also tested. The extracts from insects that are destructive of wheat grain and stored wheat products showed much higher amylase activities as compared to the other insect species that do not attack wheat and wheat products. On the basis of the effectiveness with which the three albumin fractions inhibit their activities, the amylase preparations tested were divided into susceptible, partially susceptible and resistent. Susceptible amylases, inhibited by any of the three albumin fractions, were found mainly in insects that attack wheat and in marine species. Partially susceptible amylases, inhibited by only one or two of the three albumin fractions, were present in a few avain and mammalian species including man. Resistent amylases were largely distributed in cereal, avian and mammalian species as well as in insect species that do not usually attack wheat grain or wheat flour products. At no stage of development, wheat alpha-amylase was inhibited by the albumin fractions from the mature kernel. The 12 500 dalton albumin fraction was the most effective in inhibiting insect amylases, but it was inactive toward avian and mammalian amylases. The 24 000 dalton albumin fraction was the most effective in inhibiting amylases from marine avian and mammalian species and inhibited as much as 33 amylases over 66 different amylases tested. It is suggested that protein inhibitors of amylase contributed to natural selection of polyploid wheats by giving some insect resistence to such wheats, even though some insect species were able to overcome this biochemical defense toa large degree by producing higher amylase activities.

High-performance liquid chromatographic determination of polyamines in milk as their 9-fluorenylmethoxycarbonyl derivatives using a column-switching technique.

A high-performance liquid chromatographic method for the determination of polyamines in milk is milk is described. Polyamines were extracted in perchloric acid and derivatized with 9-fluorenylmethoxycarbonyl chloride (FMOC-Cl). The excess of reagent was reacted with aspartic acid before the analysis on a column-switching system. Linearity of derivatization was calculated for each amine and the coefficient of regression ranged from 0.994 to 0.999. Chromatographic separation of FMOC-polyamines was achieved with a gradient elution programme of water-acetonitrile. The correlation coefficients of the standard curves in the concentration range from 0.5 to 5 nmol ml-1 were higher than 0.991. The repeatability of the method, expressed as R.S.D. for each polyamines ranged from 3.0 to 8.6%. The percent mean recoveries at 1 nmol ml-1 spiking level were 49 +/- 3, 58 +/- 5, 61 +/- 5 and 48 +/- 4 for putrescine, cadaverine, spermidine and spermine, respectively. The limit of detection, calculated on the basis of three times signal-to-noise ratio, was 50 pmol ml-1 for each polyamine.

Identification of species in animal feedstuffs by polymerase chain reaction--restriction fragment length polymorphism analysis of mitochondrial DNA.

Restriction site analysis of Polymerase Chain Reaction (PCR) products of cytochrome b mitochondrial DNA was applied to identify species in meat meal and animal feedstuffs. PCR was used to amplify a variable region of cytochrome b mitochondrial DNA gene. Species differentiation was determined by digestion of the obtained 359 bp amplicon with restriction enzymes, which generated species-specific electrophoresis patterns; the sequencing of PCR products was used as confirming analysis. PCR-RFLP analysis revealed the presence of meat meal in animal feedstuffs and distinguished species of interest. The results supported the application of the method in control measures which should be adopted for meat-meal-based animal feed, as suggested by EU law. As a technical improvement, to simplify the analysis, the number of enzymes presented in this study for the detection of different species was smaller than others described in the literature; discrimination between ruminant and nonruminant species and between mammalian and poultry species was possible with few digestions.

Racemization kinetics of aspartic acid in fish material under different conditions of moisture, pH, and oxygen pressure.

The racemization kinetics of aspartic acid in heat-treated whole herring have been studied under conditions of treatment comparable to those that may occur in processing of fish meal. D-Aspartic acid content in the samples has been measured by RP-HPLC with precolumn automatic derivatization. The major parameters affecting the rate of racemization of aspartic acid k(Asp) have been demonstrated to be temperature (elevation of temperature from 95 to 120 degrees C resulted in an increase of k(Asp) from 0.46 to 3.39x10(-3) min(-1)), moisture of the raw material (reduction of the moisture content of the raw material from 80 to 15% lowered k(Asp) measured at 95 degrees C from 0.46 to 0.06x10(-3) min(-1)), and to a lesser extent, pH (k(Asp) at 95 degrees C was lowered from 0.46 to 0.37x10(-3) min(-1) following a decrease of pH from 7.0 to 4.0). No significant effects on the racemization rate of aspartic acid was observed for reducing the oxygen pressure to 0.8%. The results from the present study show that the content of D-aspartic acid in fish material is a function of heat exposure and may be used to predict the thermal history of fish meal.

Adaptation of the domestic chicken, Gallus domesticus, to continuous feeding of albumin amylase inhibitors from wheat flour as gastro-resistant microgranules.

Albumin amylase inhibitors were extracted from wheat flour, precipitated by salting out the extract with ammonium sulphate, and enclosed in cellulose-coated microgranules resistant to the peptic action in the chicken gizzard. Continuous intake of gastro-resistant wheat albumins significantly (P less than 0.01) depressed chicken growth rate, whereas native wheat albumins did not show such an effect. After 4 weeks of treatment, treated chickens showed a growth rate identical to that of control chickens thus showing that an adaptation to the presence of wheat albumins in the diet had occurred. Treated chickens also showed pancreas hypertrophy and a number of histological changes in the pancreas indicating degenerative processes in progress. Moreover, in treated chickens the production of pancreatic amylase was markedly increased (P less than 0.02), whereas pancreatic protease activity was less affected. The data obtained suggest that the synthesis of pancreatic amylase in chicken is under some homeostatic control of alpha-amylase in the intestine.

The control of risk in animal husbandry primary production.

This paper describes the nutritional aspects, including drinking water, that produce a negative influence on safety, health, nutritional and technological properties of food of animal origin, such as dairy and meat products, eggs, fish, and including processed and manufactured products. The purpose was to define an overview concerning the main aspects of food safety and to produce guidelines that best fit the different breeding systems, aiming to prevent health risk to consumers from fraudulent practices that should be avoided with the application of current rules in animal husbandry.

Determination of astaxanthin stereoisomers and colour attributes in flesh of rainbow trout (Oncorhynchus mykiss) as a tool to distinguish the dietary pigmentation source.

The presence of carotenoids in animal tissue reflects their sources along the food chain. Astaxanthin, the main carotenoid used for salmonid pigmentation, is usually included in the feed as a synthetic product. However, other dietary sources of astaxanthin such as shrimp or krill wastes, algae meal or yeasts are also available on the market. Astaxanthin possesses two identical asymmetric atoms at C-3 and C-3' making possible three optical isomers with all-trans configuration of the chain: 3S,3'S, 3R,3'S, and 3R,3'R. The distribution of the isomers in natural astaxanthin differs from that of the synthetic product. This latter is a racemic mixture, with a typical ratio of 1:2:1 (3S,3'S:3R,3'S:3R,3'R), while astaxanthin from natural sources has a variable distribution of the isomers deriving from the different biological organism that synthesized it. The high-performance liquid chromatographic (HPLC) analysis of all-trans isomers of astaxanthin was performed in different pigment sources, such as red yeast Phaffia rhodozyma, alga meal Haematococcus pluvialis, krill meal and oil, and shrimp meal. With the aim to investigate astaxanthin isomer ratios in flesh of fish fed different carotenoid sources, three groups of rainbow trout were fed for 60 days diets containing astaxanthin from synthetic source, H. pluvialis algae meal and P. rhodozyma red yeast. Moreover, the distribution of optical isomers of astaxanthin in trout purchased on the Italian market was investigated. A characteristic distribution of astaxanthin stereoisomers was detected for each pigment sources and such distribution was reproduced in the flesh of trout fed with that source. Colour values measured in different sites of fillet of rainbow trout fed with different pigment sources showed no significant differences. Similarly, different sources of pigment (natural or synthetic) produced colour values of fresh fillet with no relevant or significant differences. The coefficient of distance computed amongst the feed ingredient and the trout fillet astaxanthin stereoisomers was a useful tool to identify the origin of the pigment used on farm.

Effect of temperature and diet composition on residue depletion of oxytetracycline in cultured channel catfish.

Oxytetracycline is an antibacterial agent widely used in fish farming. The normal method of administration of oxytetracycline to the fish is to mix the drug into the feed. As a consequence, the concentration of the drug in feed, together with the preparation and the composition of feed, can influence the disposition of the drug itself. An experimental study was carried out to evaluate the residue depletion of oxytetracycline from muscle tissue of channel catfish (Ictalurus punctatus) fed different medicated diets. Three hundred channel catfish were randomly divided into six tanks (50 fish per tank), maintained at water temperatures of 18 degrees C (three tanks) and 23 degrees C (three tanks). The animals were fed with three diets, differing in their energy content and composition, for the duration of the experiment oxytetracycline was added to the diets at a level of 7500 mg kg-1 for 7 d. After cessation of the treatment, five fish from each tank were killed on days 1, 3, 8, 13, 18, 24, 30, 35 and 40. Oxytetracycline residues in muscle tissue were determined by high-performance liquid chromatography. The results indicate that the energy level and chemical composition of the medicated diets administered to channel catfish influence oxytetracycline disposition in fish, and that temperature is an important factor in conditioning the reported dietary effects. Therefore, formulation of specific diets to administer drugs to farmed fish could assure better bioavailability of the chemotherapeutant and shorter withdrawal times.

High-performance liquid chromatographic determination of oxytetracycline in channel catfish (Ictalurus punctatus) muscle tissue.

A high-performance liquid chromatographic method for the determination of oxytetracycline in channel catfish muscle tissue is presented. Oxytetracycline is extracted three times from muscle tissue with an ethylenediaminetetraacetic acid disodium salt-McIlvaine buffer (pH 4.0) by using an Ultra Turrax. Analysis is carried out by using high-performance liquid chromatography and an acetonitrile-oxalic acid (0.05 mol 1(-1), pH 2.2) mixture (14 + 86, v/v) is used as mobile phase. Oxytetracycline is separated on a Lichrosorb RP-8 125 x 4.0 mm i.d. column and ultraviolet detection at 355 nm is used. The limit of quantification is 10 ng g-1 and the linearity, tested in the spiking range 20-500 ng g-1, is 0.9997. Recovery from muscle spiked at 20, 50, 100, 200 and 500 ng g-1 levels is in the range 70-80%. Precision, expressed as percentage relative standard deviation, is below 7%. The method is applied to muscle tissue from channel catfish fed on a medicated diet.