ALBERT

Notes: Abstract This study investigated the biotransformation pathways of 1,1,2,2-tetrachloroethane (1,1,2,2-TeCA) in the presence of chloroethenes (i.e. tetrachloroethene, PCE; trichloroethene, TCE) in anaerobic microcosms constructed with subsurface soil and groundwater from a contaminated site. When amended with yeast extract, lactate, butyrate, or H2 and acetate, 1,1,2,2-TeCA was initially dechlorinated via both hydrogenolysis to 1,1,2-trichloroethane (1,1,2-TCA) (major pathway) and dichloroelimination to dichloroethenes (DCEs) (minor pathway), with both reactions occurring under sulfidogenic conditions. In the presence of only H2, the hydrogenolysis of 1,1,2,2-TeCA to 1,1,2-TCA apparently required the presence of acetate to occur. Once formed, 1,1,2-TCA was degraded predominantly via dichloroelimination to vinyl chloride (VC). Ultimately, chloroethanes were converted to chloroethenes (mainly VC and DCEs) which persisted in the microcosms for very long periods along with PCE and TCE originally present in the groundwater. Hydrogenolysis of chloroethenes occurred only after highly reducing methanogenic conditions were established. However, substantial conversion to ethene (ETH) was observed only in microcosms amended with yeast extract (200 mg/l), suggesting that groundwater lacked some nutritional factors which were likely provided to dechlorinating microorganisms by this complex organic substrate. Bioaugmentation with an H2-utilizing PCE-dechlorinating Dehalococcoides spp. -containing culture resulted in the conversion of 1,1,2,2-TeCA, PCE and TCE to ETH and VC. No chloroethanes accumulated during degradation suggesting that 1,1,2,2-TeCA was degraded through initial dichloroelimination into DCEs and then typical hydrogenolysis into ETH and VC.

Notes: Abstract Microcosm experiments were conduced in which the surface of marine sediment was contaminated with naphthalene and subjected to either of three different bioremediation schemes, i.e., biostimulation (BS) by supplementing with slow-release nitrogen and phosphorus fertilizers, bioaugmentation (BA) by inoculating with Cycloclasticus sp. E2, an aromatics-degrading bacterium identified to play an important role for aromatic-hydrocarbon degradation in marine environments and combination (CB) of BS and BA. These three schemes were found to be similarly effective for removing naphthalene, while naphthalene disappearance in sediment without any treatment (WT) was slower than those in the treated sediments. Shifts in bacterial populations during and after bioremediation were analyzed by denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments. It was found that the Cycloclasticus rRNA type occurred as the strongest bands in the course of naphthalene degradation. Clustering analysis of DGGE profiles showed that bacterial populations in the WT, BS and CB sediments differed consistently from those in the uncontaminated control, while the profile for the BA sediment was finally included in the cluster for uncontaminated control sediments after a 150-day treatment. The results suggest that bioaugmentation with ecologically competent pollutant-degrading bacteria is an ecologically promising bioremediation scheme.

Notes: Abstract A short rod shaped, gram-negative bacterium strain Burkholderia sp. FDS-1 was isolated from the sludge of the wastewater treating system of an organophosphorus pesticides manufacturer. The isolate was capable of using fenitrothion as the sole carbon source for its growth. FDS-1 first hydrolyzed fenitrothion to 3-methyl-4-nitrophenol, which was further metabolized to nitrite and methylhydroquinone. The addition of other carbon source and omitting phosphorus source had little effect on the hydrolysis of fenitrothion. The gene encoding the organophosphorus hydrolytic enzyme was cloned and sequenced. The sequence was similar to mpd, a gene previously shown to encode a parathion-methyl-hydrolyzing enzyme in Plesiomonas sp. M6. The inoculation of strain FDS-1 (106 cells g−1) to soil treated with 100 mg fenitrothion emulsion kg−1 resulted in a higher degradation rate than in noninoculated soils regardless of the soil sterilized or nonsterilized. These results highlight the potential of this bacterium to be used in the cleanup of contaminated pesticide waste in the environment.

Notes: Abstract Growth of Delftia acidovorans MC1 on 2,4-dichlorophenoxyacetic acid (2,4-D) and on racemic 2-(2,4-dichlorophenoxy)propanoic acid ((RS)-2,4-DP) was studied in the perspective of an extension of the strain’s degradation capacity at alkaline pH. At pH 6.8 the strain grew on 2,4-D at a maximum rate (μmax) of 0.158 h−1. The half-maximum rate-associated substrate concentration (Ks) was 45 μM. At pH 8.5 μmax was only 0.05 h−1 and the substrate affinity was mucher lower than at pH 6.8. The initial attack of 2,4-D was not the limiting step at pH 8.5 as was seen from high dioxygenase activity in cells grown at this pH. High stationary 2,4-D concentrations and the fact that μmax with dichlorprop was around 0.2 h−1 at both pHs rather pointed at limited 2,4-D uptake at pH 8.5. Introduction of tfdK from D. acidovorans P4a by conjugation, coding for a 2,4-D-specific transporter resulted in improved growth on 2,4-D at pH 8.5 with μmax of 0.147 h−1 and Ks of 267 μM. Experiments with labeled substrates showed significantly enhanced 2,4-D uptake by the transconjugant TK62. This is taken as an indication of expression of the tfdK gene and proper function of the transporter. The uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) reduced the influx of 2,4-D. At a concentration of 195 μM 2,4-D, the effect amounted to 90% and 50%, respectively, with TK62 and MC1. Cloning of tfdK also improved the utilization of 2,4-D in the presence of (RS)−2,4-DP. Simultaneous and almost complete degradation of both compounds occurred in TK62 up to D = 0.23 h−1 at pH 6.8 and up to D = 0.2 h−1 at pH 8.5. In contrast, MC1 left 2,4-D largely unutilized even at low dilution rates when growing on herbicide mixtures at pH 8.5.

Notes: Abstract The aim of this paper was to evaluate the use of different bulking agents in different ratios as a means to control, optimise and eventually reduce the duration of the thermophilic period in two-phase olive oil mill sludge (OOMS) composting. The bulking agents used were: (i) olive tree leaves (OTL), (ii) olive tree shredded branches (OTB) and (iii) woodchips (WDC). The selection of these materials was based on their abundance and availability on the island of Crete, the southernmost point of Greece. The ratios studied were: Pile 1, OOMS:OTL in 1:1 v/v; Pile 2, OOMS:WDC in 1:1.5 v/v; Pile 3, OOMS:OTL in 1:2 v/v; Pile 4, OOMS:OTL:OTB in 1:1:1 v/v; and Pile 5, OOMS:OTL:OTB in 1:1:2 v/v. The composting system used was that of windrows with the volume of each pile approximately 20–25 m3. The experiments took place over two consecutive years. A composting turner was used and turnings were performed at one and two week intervals. In each pile a variety of physiochemical parameters were monitored. Temperature remained high in all five trials. Piles 1, 2, 3, 4 and 5 temperatures recorded values of above 50 °C for 106, 158, 160, 175 and 183 days, respectively. Volumes were reduced by approximately 67%, 62%, 63%, 80% and 84%, respectively. Temperature remained high, mainly due to the presence in large amounts of oily substances which during their complete oxidation release important amounts of energy and aid the cometabolism of more stable molecules such as lignin. This process is better described as the slow “burning” of a “fuel” mixture in an “engine” than composting. This approach is based on the extensive similarities of this process to that of crude oil sludge or similar waste composting.

Notes: Abstract Metal cyanides are significant contaminants of many soils found at the site of former industrial activity. In this study we isolated bacteria capable of degrading ferric ferrocyanide and K2Ni(CN)4. One of these bacteria a Rhodococcus spp. was subsequently used to bioaugment a minimal medium broth, spiked with K2Ni(CN)4, containing 1 g of either an uncontaminated topsoil or a former coke works site soil. Degradation of the K2Ni(CN)4 was observed in both soils, however, bioaugmentation did not significantly impact the rate or degree of K2Ni(CN)4 removal. Statistical analysis of denaturing gradient gel electrophoresis profiles showed that the topsoil bacterial community had a higher biodiversity, and its structure was not significantly affected by either K2Ni(CN)4 or bioaugmentation. In contrast, profiles from the coke works site indicated significant changes in the bacterial community in response to these additions. Moreover, in both soils although bioaugmentation did not affect rates of biodegradation the Rhodococcus spp. did become established in the communities in broths containing both top and coke works soil. We conclude that bacterial communities from contaminated soils with low biodiversity are much more readily perturbed through interventions such as contamination events or bioaugmentation treatments and discuss the implications of these findings for bioremediation studies.

Notes: Abstract To date, no pure bacterial cultures that could degrade acetochlor have been described. In this study, one strain of microorganism capable of degrading acetochlor, designated as LCa2, was isolated from acetochlor-contaminated soil. The strain LCa2 is Pseudomonas oleovorans according to the criteria of Bergey’s manual of determinative bacteriology and sequence analysis of the partial 16S rRNA gene. Optimum growth temperature and pH were 35 °C and 8.0, respectively. The strain could degrade 98.03% of acetochlor treated at a concentration of 7.6 mg l−1 after 7 days of incubation and could tolerate 200 mg l−1 of acetochlor. When the acetochlor concentration became higher, the degradation cycle became longer. The acetochlor biodegradation products were identified by GC–MS based on mass spectral data and fragmentation patterns. The main plausible degradative pathways involved dechlorination, hydroxylation, N-dealkylation, C-dealkylation and dehydrogenation.

Notes: Abstract The present work attempts to ascertain the efficacy of low cost technology (in our case, composting) as a bioremediation technique for reducing the hydrocarbon content of oil refinery sludge with a large total hydrocarbon content (250–300 g kg−1), in semiarid conditions. The oil sludge was produced in a refinery sited in SE Spain The composting system designed, which involved open air piles turned periodically over a period of 3 months, proved to be inexpensive and reliable. The influence on hydrocarbon biodegradation of adding a bulking agent (wood shavings) and inoculation of the composting piles with pig slurry (a liquid organic fertiliser which adds nutrients and microbial biomass to the pile) was also studied. The most difficult part during the composting process was maintaining a suitable level of humidity in the piles. The most effective treatment was the one in which the bulking agent was added, where the initial hydrocarbon content was reduced by 60% in 3 months, compared with the 32% reduction achieved without the bulking agent. The introduction of the organic fertiliser did not significantly improve the degree of hydrocarbon degradation (56% hydrocarbon degraded). The composting process undoubtedly led to the biodegradation of toxic compounds, as was demonstrated by ecotoxicity tests using luminescent bacteria and tests on plants in Petri dishes.

Notes: Abstract The two-tank accelerator/aerator modification of activated sludge significantly increases the biodegradation of hydrocarbons requiring initial monooxygenation reactions, such as phenol and 2,4-dichlorophenol (DCP). The small accelerator tank has a controlled low dissolved oxygen (DO) concentration that can enrich the biomass in NADH + H+. It also has a very high specific growth rate (μacc) that up-regulates the biomass’s content of the monooxygenase enzyme. Here, we develop and test the ACCEL model, which quantifies all key phenomena taking place when the accelerator/aerator system is used to enhance biodegradation of hydrocarbons requiring initial monooxygenations. Monooxygenation kinetics follow a multiplicative relationship in which the organic substrates (phenol or DCP) and DO have separate Monod terms, while the biomass’s content of NADH + H+ has a first-order term. The monooxygenase enzyme has different affinities (K values) for phenol and DCP. The biomass’s NADH + H+ content is based on a proportioning of NAD(H) according to the relative rates of NADH + H+ sources and sinks. Biomass synthesis occurs simultaneously through utilization of acetate, phenol, and DCP, but each has its own true yield. The ACCEL model accurately simulates all trends for one-tank and two-tank experiments in which acetate, phenol, and DCP are biodegraded together. In particular, DCP removal is affected most by DOacc and the retention-time ratio, Θacc/Θtotal. Adding an accelerator tank dramatically increases DCP removal, and the best DCP removal occurs for 0.2 〈 DOacc  〈 0.5 mg/l and 0.08 〈 Θacc/Θtotal 〈 0.2. The rates of phenol and DCP utilization follow the multiplicative relationship with a maximum specific rate coefficient proportional to μacc. Finally, μacc increases rapidly for Θacc/Θtotal 〈 0.25, acetate removal in the accelerator fuels the high μacc, and the biomass’s NADH + H+ content increases very dramatically for DOacc 〈 0.25 mg/l.

Notes: Abstract An outdoor experiment was conducted to study the competitionbetween Sulphate Reducing Bacteria (SRB) and Methanogenic Archaea (MA) in anaerobictreatment phase of tannery wastewater treatment in pilot-scale Advanced FacultativePond (AFP). The relative electron flow towards sulphate reduction was higher (59–83%) than towardsmethanogenesis (17–41%), although the COD recovery within thereactor varied between 15 and 90%. The results also demonstrated that the flow of electrons towards SRBincreased with increase of the sulphate concentration and decrease of the COD : SO4 = ratio.

Notes: Abstract A field experiment, lasting 14 months, was carried out in order to assess the effect of organic amendment and lime addition on the bioavailability of heavy metals in contaminated soils. The experiment took place in a soil affected by acid, highly toxic pyritic waste from the Aznalcóllar mine (Seville, Spain) in April 1998. The following treatments were applied (3 plots per treatment): cow manure, a mature compost, lime (to plots having pH 〈 4), and control without amendment. During the study two crops of Brassica juncea were grown, with two additions of each organic amendment. Throughout the study, the evolution of soil pH, total and available (DTPA-extractable) heavy metals content (Zn, Cu, Mn, Fe, Pb and Cd), electrical conductivity (EC), soluble sulphates and plant growth and heavy metal uptake were followed. The study indicates that: (1) soil acidification, due to the oxidation of metallic sulphides in the soil, increased heavy metal bioavailability; (2) liming succeeded in controlling the soil acidification; and (3) the organic materials generally promoted fixation of heavy metals in non-available soil fractions, with Cu bioavailability being particularly affected by the organic treatments.

Notes: Abstract Anaerobic enrichment cultures obtained from oil fields degraded various thiophenic compounds i.e. thiophene, benzothiophene and dibenzothiophene, with the concomitant formation of sulphide using hydrogen, lactate and ethanol as possible electron donors. It was demonstrated that dibenzothiophene was converted to biphenyl. However, hydrocarbon products from benzothiophene and thiophene desulphurisation could not be detected. After further enrichment on thiophenic compounds as the sole electron acceptor, the conversion activity disappeared while homo-acetogenic bacteria became abundantly present. In order to gain stable conversions of thiophenic compounds, attempts were made to isolate the sulphide-producing bacteria. Two highly enriched cultures were obtained, which degraded thiophenic compounds, but the activity remained low and homo-acetogenesis remained dominant.

Notes: Abstract A mixed bacterial culture capable of growing in potassium-thiocyanatecontaining medium (200 mg KSCN) has been isolated from bacterial suspensions of soilsamples collected near gold mines in Kumjung (Korea). The isolates were initially characterized by metabolic profile analysis and were identified as Bacillus thermoglucosidasius,Bacillus cereus, Bacillus licheniformis, Bacillus mycoides, Brevibacteriumepidermidis, Brevibacterium otitidis, and Corynebacterium nitrilophilus.One of the seven isolates was initially characterized as Brevibacterium epidermidis,which is not known to degrade thiocyanate. However, using 16S rDNA sequencing, thisstrain was identified as a member of Klebsiella. The strain shows high similarityvalues (95.8 to 96.4%) with Klebsiella species, and the closest known relative was foundto be K. ornithinolytica ATCC 31898. The result indicates that species of the genusKlebsiella were the closest phylogenetic relatives of the investigated strain. This isthe first known report of a member of Klebsiella that is capable of utilizing thiocyanate assole source of carbon and nitrogen.

Notes: Abstract The treatment of simulated acidic wastewater (pH 2.5–5)containing sulfate (1.0–2.2 g l-1), zinc (15–340 mg l -1) and iron (57 mg l -1) was studied in a sulfate-reducing fluidized-bed reactor (FBR) at 35 °C.The original lactate feed for enrichment and maintenance of the FBRculture was replaced stepwise with ethanol over 50 days. The robustnessof the process was studied by increasing stepwise the Zn, sulfate andethanol feed concentrations and decreasing the feed pH. The following precipitation rates were obtained: 360 mg l -1 d -1 for Zn and 86 mg l -1 d -1 for Fe, with over 99.8% Zn and Fe removal, with a hydraulic retention time of 16 h. Under these conditions, 77–95% of the electrons were accepted by sulfate reduction. The alkalinity produced from ethanol oxidation increased the wastewater pH from 2.5 to 7.5–8.5. Michaelis–Menten constants (Km) determined in batch FBR experiments, were 4.3–7.1 mg l -1 and 2.7–3.5 mg l -1 for ethanol and acetateoxidation, respectively. The maximum oxidation velocities (Vmax)were 0.19–0.22 mg gVS -1 min -1 and0.033–0.035 mg gVS -1 min -1, for ethanol and acetate, respectively. In summary, the FBR process produced a good quality effluent as indicated by its low organic content and Zn and Fe concentrations below0.1 mg l -1.

Notes: Abstract This paper describes a novel bioscrubber concept for biological flue gas desulfurization, based on the recycling of a cell suspension of sulfite/sulfate reducing bacteria between a scrubber and a sulfite/sulfate reducing hydrogen fed bioreactor. Hydrogen metabolism in sulfite/sulfate reducing cell suspensions was investigated using batch activity tests and by operating a completely stirred tank reactor (CSTR). The maximum specific hydrogenotrophic sulfite/sulfate reduction rate increased with 10% and 300%, respectively, by crushing granular inoculum sludge and by cultivation of this sludge as cell suspension in a CSTR. Operation of a sulfite fed CSTR (hydraulic retention time 4 days; pH 7.0; sulfite loading rate 0.5–1.5 g SO 3 2- l-1 d-1) with hydrogen as electron donor showed that high (up to 1.6 g l-1) H2S concentrations can be obtained within 10 days of operation. H2S inhibition, however, limited the sulfite reducing capacity of the CSTR. Methane production by the cell suspension disappeared within 20 days reactor operation. The outcompetition of methanogens in excess of H2 can be attributed to CO2 limitation and/or to sulfite or sulfide toxicity. The use of cell suspensions opens perspectives for monolith or packed bed reactor configurations, which have a much lower pressure drop compared to air lift reactors, to supply H2 to sulfite/sulfate reducing bioreactors.

Notes: Abstract The enzymatic pathways of elemental sulfur and thiosulfate disproportionation were investigated using cell-free extract of Desulfocapsa sulfoexigens. Sulfite was observed to be an intermediate in the metabolism of both compounds. Two distinct pathways for the oxidation of sulfite have been identified. One pathway involves APS reductase and ATP sulfurylase and can be described as the reversion of the initial steps of the dissimilatory sulfate reduction pathway. The second pathway is the direct oxidation of sulfite to sulfate by sulfite oxidoreductase. This enzyme has not been reported from sulfate reducers before. Thiosulfate reductase, which cleaves thiosulfate into sulfite and sulfide, was only present in cell-free extract from thiosulfate disproportionating cultures. We propose that this enzyme catalyzes the first step in thiosulfate disproportionation. The initial step in sulfur disproportionation was not identified. Dissimilatory sulfite reductase was present in sulfur and thiosulfate disproportionating cultures. The metabolic function of this enzyme in relation to elemental sulfur or thiosulfate disproportionation was not identified. The presence of the uncouplers HQNO and CCCP in growing cultures had negative effects on both thiosulfate and sulfur disproportionation. CCCP totally inhibited sulfur disproportionation and reduced thiosulfate disproportionation by 80% compared to an unamended control. HQNO reduced thiosulfate disproportionation by 80% and sulfur disproportionation by 90%.

Notes: Summary Background: The knowledge of magnesium requirements of premature infants is still very limited, although it is essential for the optimal composition of suitable formulas. Aim of the study: The study concept was 1) to assess physiological magnesium balance data of healthy term infants and longitudinal results from formula-fed premature infants and 2) to deduce conclusions on the magnesium content of the formulas. Methods: Premature infants (n = 14, birth weight ≤ 1500 g, gestational age ≤ 32 weeks) were studied in conventional balance trials with 1) a semi-elemental diet (A), 2) preterm infant formula (B), and 3) infant formula (C). In addition, healthy term formula-fed (n = 11, D) and breast-fed (n = 14, E) infants were investigated. Analysis was performed by flame atomic absorption spectroscopy. Results: The median magnesium intake ranged between 4.84 mg/kg × d−1 (breast-fed infants) and 16.33 mg/kg × d−1 (premature infants). The term breast-fed infants retained nearly as much magnesium as term formula-fed infants (3.37 vs. 3.97 mg/kg × d−1), due to a low percental fecal and urinary excretion. A higher magnesium retention was observed in the premature group: A: 7.97 mg/kg × d−1, B. 5.3 mg/kg × d−1, 3.) 5.54 mg/kg × d−1. Conclusion: In view of the high percental magnesium retention in formula-fed premature infants, excessive supply should be avoided. The long-term effects of lower intakes have to be monitored.

Notes: Summary We reviewed 15 studies reporting on the fatty acid composition of colostrum lipids from 16 geographic regions: 11 European studies and one study each from Central America, the Caribbean, Australia and Asia. The contents of essential fatty acids, saturates and polyunsaturates were similar in the southern European countries Spain, Slovenia and France. Colostrum of St. Lucian women was high in saturates and low in oleic acid, reflecting a high-carbohydrate, low-fat diet. Abundant fish intake was reflected in high contents of docosahexaenoic acid and total n-3 long-chain polyunsaturated fatty acids in St. Lucia. Two French studies published with an interval of two years showed a very similar colostrum fatty acid composition, whereas two German studies obtained with an interval of 14 years showed higher docosahexaenoic acid and arachidonic acid contents in the later study, with an unchanged n-6/n-3 long-chain polyunsaturated fatty acid ratio. Studies from Spain reported a decline of α-linolenic acid in colostrum over a time period of 13 years. Colostrum of Australian women contained the lowest polyunsaturated/saturated and n-6/n-3 long-chain polyunsaturated fatty acids ratios (0.28 and 1.58) and the lowest contents of linoleic and α-linolenic acids (7.8 and 0.4 wt. %). In contrast, the contents of docosahexaenoic acid, eicosapentaenoic acid and total n-3 long-chain polyunsaturated fatty acids (0.6, 0.4 and 1.4 wt. %) were higher in Australian than in European samples. Fatty acid composition of human colostrum appears to be markedly influenced by geographic differences in maternal dietary composition.

Notes: Summary The metabolic strengths, weakness, opportunities and threats of the metabolic ability to split water brought about a proliferation of biological systems, produced a toxic oxygenic environment, and were responsible for the development of antioxidant defence mechanisms. Evolution is driven by heritable adaptions which improve environmental ‘fit’. Hence aerobic respiration, using oxygen as a nutrient, came to predominate in biological systems, and antioxidant defence mechanisms which prevent and neutralise toxic oxygen intermediates have become widespread, varied, coordinated and effective. Antioxidant defences are nor infallible however. In humans, reactive oxygen species-induced damage is associated with the ageing process, and with chronic diseases including cancer and coronary heart disease. Interestingly, some important antioxidants, including ascorbic acid and the tocopherols, cannot be synthesised by humans and must be taen in the diet. Another antioxidant, uric acid, is found in much higher concentrations in humans than in other mammals, and levels are also affected by diet. In humans, therefore, antioxidant defence against toxic oxygen intermediates is species specific and heavily influenced by nutrition. In this article, the atmospheric and metabolic changes which produced both the threat and opportunity offered by an oxygenic environment are outlined. An overview of oxygen toxicity, and adaptations to oxidative stress in terms of evolution of antioxidant defences, is presented. Finally, suggested benefits underlying our curious inability to manufacture ascorbic acid, and the possible role of uric acid in human antioxidant defence, are briefly discussed with particular reference to nutrition and toxicology.

Notes: Summary Background: Many epidemiological studies have identified a protection against cancer associated with consumption of fruit and vegetables. One factor is this protection may be the enhancement of cellular DNA repair activity by micronutrients, such as carotenoids, found in these foods. Aims of the study: To measure the capacity of lymphocytes isolated from volunteers supplemented with β-carotene, lutein or lycopene to recover from DNA damage induced in vivo by treatment with H2O2. Methods: Healthy volunteers were given supplements of lutein (15 mg/day), lycopene (15 mg/day) and βcarotene (15 mg/day), each for 1 week, the supplementation periods being separated by 3-week wash-out periods. Blood samples were taken at the beginning and end of each supplementation, and at 1 week and 3 weeks during the wash-out period. Carotenoid levels were measured in plasma. Lymphocytes were isolated and frozen. Subsequently, they were treated with 100 μM H2O2 and incubated for up to 24 h; DNA damage was measured with the comet assay (single cell gel electrophoresis) after 0, 2, 4, 8, and 24 h. Results: Increases of 2- to 3-fold in mean plasma lutein and β-carotene concentrations were seen at the end of the respective supplementation periods; they returned virtually to basal levels after wash-out. Lycopene concentrations were less affected by supplementation, and were more variable. H2O2-induced DNA strand breaks were apparently only slowly rejoined by the lymphocytes. The rejoining of breaks in the first few hours appeared substantially faster in lymphocytes following supplementation with β-carotene, but no such effect was seen with lutein. In those individuals who showed increases in lycopene concentrations, the recovery was significantly faster. Lymphocytes that were not treated with H2O2 showed a transient increase in DNA breakage to about double the background level in 2 h, presumably as a result of exposure to atmospheric oxygen; this effect, too, was relieved by supplementation with lycopene or β-carotene. Conclusions: While certain carotenoids appear to enhance recovery from oxidative damage, this is probably in fact an antioxidant protective effect against additional damage induced by atmospheric oxygen, rather than a stimulation of DNA repair.

Notes: Summary Obesity could well become the most common health problem of the 21st century. There are more opportunities to consume large quantities of food: big portions of tasty, varied food, at reasonable prices, are available everywhere. Moreover, our bodies are better adapted to combat weight loss than to combat weight gain, since for thousands of years our species evolved in circumstances where nutrients were in short supply. The response of each individual to diet and other environmental factors varies considerably, depending on the characteristics of his/her body weight control mechanism. The differentiating element in the future, especially as regards the dietary and pharmacological control of obesity, will be knowledge of an individual's possible response depending on his/her genetic background. Obesity can occur as a result of genetic or acquired changes in three main types of biochemical processes, which are the main focus of this review: a) feeding control, which determines the sensations of satiety and hunger through processes that depend on a interplay between internal signals (notably leptin) and environmental factors; b) energy efficiency, in particular the activation of thermogenesis mediated by uncoupling proteins (UCPs) that makes it possible to dissipate part of the energy contained in food as heat instead of accumulating it as fat, and c) adipogenesis, the process by which cells specialised in fat storage (adipocytes) are formed, which is controlled by an interplay of transcription factors, including memebers of the C/EBP, PPARγ and ADD families. The knowledge of a growing numbers of genes and molecules implicated in these three types of processes and of their metabolic relationships is leading toward a molecular understanding of the body weight regulatory system and is paving the way for new methods of obesity control, especially pharmacological but also nutritional and possibly involving genetic intervention.

Notes: Summary Background: Quantitative information on conversion of β-carotene to vitamin A in humans is limited. Aim of the study: Our laboratory has developed a stable isotope method for studying the conversion of β-carotene (β-C) to vitamin A. Methods: Two dosage levels (a pharmacological dose, 126.0 mg β-C-d 8, and a physiological dose, 6.0 mg β-D-d 8) were used 2.5 y apart in an adult female volunteer to study dose effects on the conversation of β-C to vitamin A. Blood samples were collected over 21 d. β-C and retinol were extracted from serum and isolated by high performance liquid chromatography. The retinol fraction was derivatized to a trimethylsilyl ether which was analyzed by gas chromatograph/mass spectrometry with electron capture negative chemical ionization. Results: The retinol-d 4 response in the circulation peaked at 24 hours after the β-C-d 8 dose, with a higher percent enrichment after the pharmacological dose than after the physiological dose. By using retinyl acetate-d 8 as the vitamin A reference, the retinol-d 4 formed from 6 mg of β-C-d 8 (11.2 μmol) was calculated to be equivalent to 1.6 mg of retinol (i. e., 3.8 mg of β-C was equivalent to 1 mg of retinol). However, the retinol-d 4 formed from 126 mg of β-C-d 8 (235 μmol) was equivalent to 2.3 mg of retinol (i. e., 55 mg β-C was equivalent to 1 mg retinol). Conclusion: These results provide evidence that it is feasibile to use stable isotope reference method to study retinol equivalence of β-C and that there may be a dose-dependence on bioconversion of β-carotene to retinol.

Notes: Summary The potsitive effect of glutamine on lymphocyte proliferation has previously been described. Its dipeptide glycyl-glutamine (GlyGln) is more stable than pure glutamine in aqueous solutions. The aim of our study was to investigate the relationship between lymphocyte proliferation and varying concentrations of glycyl-glutamine in vitro. Isolated human lymphocytes were stimulated with the mitogens phytohaemagglutinin (PHA), Concanavalin A (ConA), pokeweed mitogen (PWM), and Staphylococcus aureus (SAC). Glycyl-glutamine was added to yield final cincentrations of 0–2 mmol/l. Overall minimal concentrations of 0.01 mmol/l glycyl-glutamine were sufficient to enhance lymphocyte proliferation over baseline (glutamine-free) levels. No difference was found between concentrations in the “physiological” range of 0.4 mmol/l and very low concentrations (0.04–0.1 mmol/l) with SAC, ConA and PWM. Increasing the concentration beyond 0.4 mmol/l (up to 2.0 mmol/l) offered further gain with PHA-stimulation only. Lymphocyte proliferation under in vitro polyclonal stimulation is maintained even at very low concentrations of glycyl-glutamine. Raising the concentration above the equivalent of physiological levels does not seem to provide further benefit.

Notes: Summary Aim of the study. An allergen-reduced dietary intervention programme with strict dietary requirements was implemented over the first four months of life in an unselected population-based infant cohort and compared to a non-intervention cohort (the ZUFF study). Recommendations for the dietary programme in the intervention cohort were extended, but not strictly implemented, until the end of month six. The intervention was based on breastfeeding, a moderate whey hydrolysate formula (pHF), and delayed introduction of weaning foods with a high allergenicity. This study was a prospective, controlled, and unblinded study, the first to assess the effects of an allergen-reduced, pHF-based early nutritional programme in a broad unselected infant population. Because overall healthy development of the infant is a major objective of any nutritional programme, the study evaluated the effects of the dietary intervention on infant growth and general health status rather than specific allergic manifestations. Part I of this paper gave results for nutritional behaviour only, and Part II gives results for growth and general health status during the intervention period through the sixth month of life. Methods. Assignment of study infants was to demographically comparable intervention (Z) or control (FF) cohorts according to place of birth. In the intervention cohort (Z=564), the recommended dietary regimen was breastfeeding and – if exclusive breastfeeding was not possible – supplementation with a moderately hydrolysed, allergen-reduced infant formula (pHF). Weaning foods were delayed until four months of age or later in case of weaning foods with high allergenicity. In the control cohort (FF=566), there was no specific intervention. Imbalances between cohorts in confounding (adjuvant) factors that could influence health related outcomes were integrated as covariates into the logistic regression of the main analyses. Growth parameters included weight, length, head circumference, BMI, and Z scores (SDS). General health status was assessed by clinically significant findings in gastrointestinal, respiratory, or skin symptoms. Results. Growth at 6 weeks and at 3 and 6 months was similar for Z and FF. Significantly fewer Z than FF infants had clinically noteworthy health findings at 3 months (Z=27% versus FF=37%, odds ratio=0.63, CI=0.48–0.82) and 6 months (Z=33% versus FF=49%, odds ratio=0.51, CI=0.40–0.66). This corresponds to a 30% reduction in overall health concerns at 6 months for the intervention cohort. At 3 and 6 months, differences between cohorts in most measures of general health status were strongly influenced by a lower incidence of skin symptoms in the Z cohort. Within FF, there were fewer exclusively breastfed (eBF) infants with health problems at 3 months compared with those who were partially (pBF) or non-breastfed (nBF) (eBF=31%, pBF=40%, nBF=39%, p〈0.05). In contrast, in the Z intervention cohort, the number of infants with health concerns was similar for exclusively breastfed infants and for those in whom mother's milk was supplemented or replaced by pHF (eBF=29%, pBF=25%, nBF=26%, ns). In a subanalysis of overall health findings in infants without a family risk of allergies, there were again significantly fewer Z than FF infants with any health or any skin problem. Conclusion.An allergen-reduced dietary recommendation that includes a moderate whey hydrolysate infant formula (pHF) has no negative effects on growth parameters up to 6 months of life in an infant population unselected for atopic risk. The dietary intervention produced improvements in general health status when compared with a control cohort that received infant formula with unhydrolysed proteins (IF), and high allergenic weaning foods at an earlier age. The difference between cohorts was principally due to fewer adverse skin findings. In infants following our allergen-reduced feeding recommendation, 3-month general health status was comparable between those who were exclusively breastfed and those in whom breastfeeding was supplemented or replaced by pHF. Our results demonstrate that a pHF feeding recommendation during the first 4 to 6 months of life – when exclusive breestfeeding is not possible – is a safe and feasible regimen not only in high-allergic risk populations but in general unselected infant population. The general use of pHF formula in non- or partly breastfed infants could therefore be considered an important contribution to optimised infant nutrition.

Notes: Summary Background: The dietary supplementation with EPA (eicosapentaenoic adic; 20:5n3) and DHA (docosahexaenoic acid; 22:6n3) has been recommended because of their favourable effects on the cardiovascular system (including complications of NIDDM). Oleic acid (18:1n9) from olive oil has some analogous and complementary effects. Potential competitive relations between long-chain n-3 fatty acids (FAs) and the oleic acid would therefore mean a problem. Aim of the study: We focused primarily on the oleic acid changes in serum phospholipids (SPL) after a supplementation with EPA and DHA. Methods: Thirty-five patients with type 2 diabetes mellitus (NIDDM) were supplemented for 28 days with 1.7 g of EPA plus 1.15 g of DHA/day (as Maxepa® capsules, Seven Seas®, U. K.). After that, a 3-month wash-out control period with 21 patients followed. A fatty acid composition of serum phospholipids (SPL) was determined by capillary gas-chromatography. Values were calculated as relative percentages of all FAs. Results: After the supplementation with the Maxepa® capsules, there was a very strong increase in EPA, docosapentaenoic acid (22:5n3) and DHA content in SPL. It was followed by a stron decrease after the wash-out (all p〈0.0001). The oleic acid SPL content after the intervention significantly decreased from 10.105±0.307% (mean ±S.E.M.) to 9.082±0.276% (p〈0.0003). During the wash-out, the change was in the opposite direction (p〈0.0001). When the intervention and the wash-out periods were taken together, changes in the oleic acid were inversely correlated with changes in EPA, docosapentaenoic acid and DHA (r = −0.729; r = −0.552; r = −0.629, respectively; p〈0.0001; n = 56). On the background of the overall n-6 FA reduction, the decline in the arachidonic acid after the supplementation (p〈0.0001) and its rise after the wash-out (p〈0.0003) were similar. There were no significant changes in the saturared FA spectrum. Conclusions: Supplementation with long-chain n-3 FAs in NIDDM patients leads to the lowering of oleic acid SPL content. Whereas the reduction of the arachidonic acid may have some desirable aspects (e. g. suppression of thromboxane TxA2 or 4 series leukotriene production), the decline of the former is to be regarded as a potential problem. Therefore, the search for optimally balanced blends of n-3 polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) seems to be more promising than a supplementation with only one type of FA.

Notes: Summary Background Present animal models used to emulate type 2 diabetes may not accurately reflect the metabolic changes that occur in humans. Aims of the study The purpose of this research was to evaluate diets reported to induce insulin resistance and impaired glucose metabolism in rats as a potentially useful model for studying type 2 diabets. Methods Three groups of male Sprague Dawley rats (n=7) were fed either a control diet, based on AIN recommendations (53% cornstarch, 10% sucrose and 7% soybean oil), a high fat diet (25% soybean oil, 35% cornstarch) or a high fructose diet (53% fructose, 10% sucrose) for a 3 month period. Glucose tolerance tests were carried out in week 3 and week 9 of the experiment. At the termination of the experiment, serum insulin, glucose, cholesterol and triacylglycerols were measured. Glucose incorporation into glycogen and glycogen synthase activity were measured in soleus muscles. Results Similar weight gain was observed for all three groups of rats. Glucose tolerance curves and fasting glucose levels were not significantly different at any time point in the experiment. Insulin levels were unchanged for the controls (171±21 pM), high fructose (164±16 pM) and high fat (181±30 pM) diets. Fasting serum triacylglycerols and cholesterol levels were not significantly elevated by dietary treatment. In soleus muscles, rats on all three diets had a significant increase in glycogen synthesis in response to insulin, but synthesis was similar in all three groups. Glycogen synthase activity was also not significantly affected by long-term dietary intervention. Conclusions In this study, healthy Sprague Dawley rats fed high fat or high fructose diets for 3 months adapted to the nutritional intervention without developing classical signs of insulin resistance and impaired glucose tolerance.

Notes: Summary Background Bone mineral density (BMD) is largely genetically determined and this influence is most powerful in the period of rapid skeletal development in childhood and late adolescence but environmental factors such as exercise and dietary calcium intake may influence up to 20%. Aims of the study The aims of the study were to examine healthy late adolescent females for the effects and benefits of a high calcium intake from dairy product foods on bone mineral density, body composition, lipids and biochemistry. The secondary aim is determine whether a high intake of dairy product foods in the diet is acceptable for this age group long term. Methods Ninety-one teenage girls who participated in a two-year randomised controlled study on the effect of dairy food supplementation on dietary patterns, body composition and bone density in post-pubertal teenage girls were approached one year after the cessation of the study to determine the effects of the cessation of dairy supplements on bone mineral density, dietary habits, biochemical markers, body composition and blood lipids. Bone mineral density and bone mineral content were assessed at the hip, spine and total body. Anthropometric data were collected, and exercise, Tanner, dietary assessment, preference and compliance questionnaires were administered. Lipid profiles, hydroxyproline excretion and urinary calcium and sodium excretion measurements were performed. Results There were no significant differences between the 2 groups for height, weight, lean and fat mass. The supplemented group had significantly higher calcium, phosphorus and protein intake during the supplementation period (p〈0.001). No differences were seen between the groups 12 months after supplementation finished. There were no significant differences in exercise level, preference or acceptability of dairy products or in the lipids and bone markers between baseline the end of supplementation and 1 year follow-up. There was a significant increase in trochanter (4.6%), lumbar spine (1.5%) and femoral neck (4.8%) BMD (p〈0.05) in the high calcium group at the end of supplementation. There was an increase in bone mineral content at the trochanter (p〈0.05) and lumbar spine; however the latter was not statistically significant, in the high calcium group at the end of supplementation. There was no difference in vertebral height or width at any stage of the study, indicating no influence on bone size. Conclusions In this 3 year study (2 years of supplementation, 1 year follow-up), teenage girls, aged 15–18 years, were able to significantly increase their BMD at the trochanter, femoral neck and lumbar spine when supplemented with dairy product foods to a mean calcium intake of 1160 mg/d. There was also an effect seen on the BMC particularly at the trochanter and to a lesser extent at the lumbar spine. The dietary calcium intake achieved did not adversely affect body weight, fat and lean mass or blood lipid profiles. Twelve months after the supplementation finished the girls had returned to their baseline diet, indicating self-selection of a high dairy product diet may be hard to achieve.

Notes: Summary Alpha-tocopherol occurs in nature as a single stereoisomer (RRR) while synthetic vitamin E is a mixture of eight stereoisomers (all-racemic, all-rac). The presently accepted ratio of biopotency (RRR: all-rac) is 1.36, based on the fetal resorption test in rats. This ratio has been disputed for humans. Clinical endpoint studies in humans are lacking, but plasma responses to RRR-and all-rac were measured in bioavailability studies. In nine studies comparing unlabeled forms, the ratio of plasma parameters (AUC, Cmax or steady-state concentration) concurred with the accepted ratio of biopotency within accepted bounds of equivalence. Four recent studies with simultaneous application of trideutero-RRR and hexadeutero-all-rac resulted in ratios of up to 2 for plasma, and of ≈ 2.7 and ≈ 3.4 for α-CEHC (a urinary metabolite) and umbilical cord plasma, respectively. Because these results have been widely assumed to reflect the difference in biopotency, this has prompted a proposal to the Food and Nutrition Board, National Academy of Sciences, USA to change the biopotency factor to 2:1. We challenge the validity of bioavailability data in lieu of clinical endpoints. Because RRR and all-rac are not chemically identical and differ in plasma and tissue kinetics and metabolism, the ratio of bioavailability parameters does not reflect the ratio of biopotency. This needs to be determined in adequately designed studies using clinical and biochemical endpoints. Until such studies have been performed it does not appear prudent to exchange the presently accepted ratio based on valid bioassays, albeit in a model animal, for another that is based on erroneous conclusions from human studies.

Notes: Summary Background Gastric emptying of non-nutrient liquids usually lacks the presence of an initial delay phase (lag phase), and so it has been considered to be monoexponential with an initial rapid phase followed by a slower emptying phase. However a lag phase in the gastric emptying of liquids can be found if there is a high caloric density in the liquid meal. Aims of the study To characterise with stable isotopes the presence of a lag phase in the gastric emptying of non-sold meals. Methods Healthy volunteers ingested a low caloric liquid meal (345 KJ/200 mL) (LCLM), a high caloric liquid meal (1135 KJ/180 mL) (HCLM) or a semisolid meal (1403 KJ/500 mL) (SSM). Test meals were labelled with 13C-acetate. Breath samples were collected for 13〉CO2 measurement and data were fitted to a power exponential function. Results Non-solid meals can have different behaviour related to the initial emptying. The presence of a lag phase in the gastric emptying of liquids was not masked by the processing of the tracer previous to its detection in breath. While the LCLM and SSM showed a rapid initial emptying phase (no lag phase), the HCLM has an initial slow emptying phase. The slower gastric emptying of the HCLM compared to the SSM was related to the presence of a lag phase in the gastric emptying of the HCLM. Conclusions The 13C-acetate breath test is very accurate to identify and study the lag phase if present of liquid meals.

Notes: Summary Background Animal studies have demonstrated dramatic changes in the intestinal flora during total enteral (TEN) or parenteral (TPN) nutrition. Aims of the study To assess the impact of TEN and TPN on human intestinal microflora. Methods Eight patients on fiber-free TEN, five patients on TPN, and ten controls were studied. Fecal bacteria were identified and numbered (logCFU/g feces), and fecal short-chain fatty acids (SCFAs) were measured in stool samples, by gas-liquid chromatography. Results In TEN patients, compared to controls (P〈0.01), aerobes were increased (8.46±0.24) while anaerobes were decreased (5.79±0.84). In TPN patients, both aerobes and anaerobes were decreased compared to controls (5.64±0.27 and 5.31±1.09 respectively, P〈0.01). Total SCFAs were lower in TPN patients than in TEN patients (48.3±16.6 vs 118.6±24.1 mmol/kg, P〈0.05). Conclusions Both TPN and TEN induce modifications in the intestinal microflora. During TPN, a homogeneous decrease occurs in both aerobic and anaerobic bacteria. TEN decreases only anaerobic bacteria, while aerobic bacteria are increased. This imbalance may play a role in the pathophysiology of TEN-induced diarrhea.

Notes: Summary The nutritional patterns of Paleolithic humans influenced genetic evolution during the time segment within which defining characteristics of contemporary humans were selected. Our genome can have changed little since the beginnings of agriculture, so, genetically, humans remain Stone Agers – adapted for a Paleolithic dietary regimen. Such diets were based chiefly on wild game, fish and uncultivated plant foods. They provided abundant protein; a fat profile much different from that of affluent Western nations; high fibre; carbohydrate from fruits and vegetables (and some honey) but not from cereals, refined sugars and dairy products; high levels of micronutrients and probably of phytochemicals as well. Differences between contemporary and ancestral diets have many pathophysiological implications. This review addressed phytochemicals and cancer; calcium, physical exertion, bone mineral density and bone structural geometry; dietary protein, potassium, renal acid secretion and urinary calcium loss; and finally sarcopenia, adiposity, insulin receptors and insulin resistance. While not, yet, a basis for formal recommendations, awareness of Paleolithic nutritional patterns should generate novel, testable hypotheses grounded in evolutionary theory and it should dispel complacency regarding currently accepted nutritional tenets.

Notes: Summary Background: Epidemiological studies report an inverse relationship between intake of the B vitamine folic acid and colon cancer. Folate is important for DNA synthesis and repair. Moreover, the production of S-adenosylmethionine (SAM), essential for normal DNA methylation and gene expression, is dependent on folic acid. Folate deficiency may increase the risk of malignant transformation by perturbing these pathways. Aims of the study: The principal aim of this study was to determine the effects of folate deficiency on DNA stability and DNA methylation in rat colonocytes in vivo. As the metabolic pathways of folate and other dietary methyl donors are closely linked, the effects of methionine and choline deficiency were also evaluated. Methods: Male Hooded-Lister rats were fed a diet deficient in folic acid, or in methionine and choline, or in folate, methionne and choline for 10 weeks. DNA strand breakage and misincorporated uracil were determined in isolated colonocytes using alkaline single cell gel electrophoresis. Global DNA methylation was measured in colonic scrapings. Folate was measured in plasma, erythrocyte and liver samples. Results: Methyl donor deficiency induced DNA strand breakage in colonocytes isolated from all experimental groups. Uracil levels in colonocytes DNA remained unchanged compared with controls. DNA methylation was unaffected either by folate and/or methionine and choline depletion. Rats fed a folate-deficient diet had less folate in plasma, red blood cells and liver than controls. Conclusions: Folate and methyl deficiency in vivo primarily afects DNA stability in isolated colonocytes of rats, without affecting overall DNA methylation.

Notes: Summary Background: Metallothionein (MT)# synthesis can be stimulated in many organs not only by various metals such as cadmium, zinc, and copper, but also by many nonmetalic compounds or experimental conditions such as oxidative stress. The latter lead to the hypothesis that MT is induced in response to free radicals formed in tissues and lipid peroxidation. Aims of the study: Whether the relationship between lipid peroxidation amd MT synthesis is a common phenomenon also valid for lipid peroxidation induced by dietary factors such as chronic vitamin E inadequacy and autoxidation products of polyenoic fatty acids derived from thermally oxidized oil was investigated in the presence study. Methods: The relationship between the induction of metallothionein isoforms I and II (MT-I and MT-II) in response to diet-induced lipid peroxidation using a rat model system in which lipid peroxidation was examined in vivo by chronic vitamin E inadequacy or by administration of lipid peroxidation products from a thermally treated polyenoicrich oil with either basal (dietary zinc concentration: 48 mg/kd; experiment 1) or Zn-stimulated MT levels (dietary zinc concentration: 305 mg/kd; experiment 2) was studied. In both experiments, growing male rats were fed diet containing either a fresh or a thermally treated soybean oil with deficient of sufficient amounts of vitamin E (14 and 11 vs. 648 and 560 mg α-tocopherol equivalents per kg diet) over 40 days according to a bifactorial experimental design. Plasma and liver concentrations of tocopherols and hepatic levels of thiobarbituric acid-reacitve substances (TBARS) were measured by high performance liquid chromatography. MT isoform concentrations in rat liver were isolated and quantified by ion-exchange high performance liquid chromatography and atomic absorption spectrometry. Results: Irrespective of the zinc supply, rats receiving inadequate amounts of vitamin E with the diet had markedly lower plasma and liver concentrations of α-tocopherol and total tocopherols than vitamin E-sufficient rats. ANOVA also revealed an interaction between the diet factors vitamin E and oil on tocopherols in plasma and liver of rats from both experiments. In experiment 1, where rats received normal amounts of dietary zinc, ingestion of the thermally treated oil impaired the tocopherol status compared to the treatment with the fresh oil, although this effect was only obvious in the vitamin E-deficient groups. In experiment 2, where rats received excessive amounts of zinc, the thermally treated oil did not contribute to a reduction of the tocopherol status in plasma and liver. In both experiments a significant increase in TBARS level, indicative of lipid peroxidation, was observed in the liver at chronic vitamin E inadequacy, but no effect of the oil was observed. Here, we show that the dietary treatment had some effects on the synthesis of liver metallothionein isoforms. In groups, receiving normal amounts of zinc, there was a significant interaction between the dietary treatments on the levels of MT-I and MT-II in liver. Chronic vitamin E inadequacy which was accompanied by diminisched tocopherol levels in liver induced the synthesis of MT-I and MT-II. When vitamin E inadequacy was combined with the ingestion of a thermally treated polyenoic acid-rich oil hepatic levels of MT-I and MT-II remained low. In experiment 2, where rats were fed the high zinc diet, vitamin E inadequacy caused an increase of hepatic MT-I level just as in experiment 1, although this MT stimulating effect was irrespective of the oil. For MT-II there was a 43% increase in the vitamin E-deficient group fed the fresh oil compared to all the other groups, although this effect was not statistically significant. The liver MT isoform response to stress was similar in rats with basal MT levels and Zn-induced liver MT levels. The failing effect of the thermally treated oil on MT levels which were stimulated by vitamin E deficiency in experiment 2 was possibly due to the low oxidation grade of the thermally treated oil. Conclusion: The present results are strongly indicative of an apparent induction of MT isoform synthesis in response to an impaired antioxidant defence system in the lipid regions of liver cells induced by vitamin E inadequacy. In contrast, thermally treated polyenoic-rich oils with a certain oxidation grade seem to restrain the induction of MT isoform synthesis under the present experimental conditions.

Notes: Summary Background Muscle triglycerides are important as a source of energy and in relation to metabolic sensitivity. However, the classic biopsy method does not distinguish intra- from extracellular fat, and their regulation by exercise and diet is largely unknown. Magnetic resonance spectroscopy (MRS) is available to assess the intramyocellular lipid (IMCL) pool non-invasively in humans. Aims of the study The aim of this work was to use sequential MRS measurements of IMCL and glycogen to explore the role of three levels of dietary fat on the replenishment of these energy stores after exercise. Methods Following 2h of exercise, two subjects (S1, S2) were fed one of three diets (15%, 40% or 70% fat energy), each on a separate occasion. IMCL and glycogen were measured by MRS in the tibialis anterior muscle before, after exercise, and at 10 and at 32 h of recovery. Results Initial IMCL concentration (mmol · kg−1:3.0 in S1 and 1.8 in S2) was reduced to 70% after exercise. The rate of replenishment was minimal with the low-fat (mmol · kg−1· 24 h−1: 0.7 and 0.0) and much higher with both higher fat diets (mmol · kg−1· 24 h−1: 3.1 and 3.2 in S1, 0.7 and 0.9 in S2). Glycogen and IMCL replenishments were inversely correlated. Conclusions IMCL and glycogen can vary acutely in response to diet after exercise. Studies are needed to determine if such variations occur within the range of ordinary diets and to clarify the functional significance of IMCL in differently active individuals.

Notes: Summary Background Recent data suggest that daily iron supplementation of iron-replete children could impair their growth. If verified for weekly iron supplementation these results would markedly complicate targeting and implementing school-based weekly iron supplementation programs. Aim of the study To ascertain the effect of weekly iron supplementation on the growth and hemoglobin status of non-anemic school-age children. Subjects and methods 73 Bolivian non-anemic school-age children randomly assigned to the treatment group (n=37; receiving supplements containing FeSO4 during 18 weeks) or the control group (n=36; receiving a placebo during the same period). Hemoglobin concentration and anthropometric measures were determined for each child at the beginning (T0) and the end (T18) of the study. Results The treatment group did not show any significant variation in hemoglobin concentration between T0 and T18 (−1.6±10.4 g/L; P=0.40) whereas the control group showed a significant decrease in hemoglobin concentration (−4.6±10.9 g/L; P=0.03). Anthropometric changes were not significantly different between the treatment and the control groups for weight, (1.63±1,11 kg vs 1.88±0.79 kg; P=0.30), height (2.35±0.94 cm vs 2.11±1.03 cm; P=0.34) or mid-upper arm circumference (0.29±0.57 cm vs 0.22±0.54 cm; P=0.64). Conclusion In our study, weekly iron supplementation of non-anemic school-age children had no negative effect on their growth while having a positive effect in preventing significant decreases in hemoglobin concentration. These results suggest that in regions where iron deficiency anemia (IDA) is prevalent, a simple and cost-effective way to control IDA in school-age children is to give weekly iron supplements to all children at school.

Notes: Summary Background: Diet restriction (DR) has been shown to extend the life spans of various laboratory animals, the mechanism may involve a decrease in oxidative stress. When determining if the total tissue defense has been altered, it is important to observe the overall direct antioxidant capacity, which consists of low molecular weight antioxidants (LMWA) and enzymes. Aim: To determine DR induced changes in total reducing power and overall direct antioxidant capacity of various mouse tissues. Methods: Young female Sabra mice were fed a 60% food restricted diet for 40 days (DR group). Organs of the DR group and of ad libitum (AL) fed controls were then dissected and examined. A cyclic voltammetry method was used to quantify the total reducing power, which correlates with the overall LMWA activity. Specific LMWA were identified by HPLC-ECD. Superoxide dismutase activity and H2O2 degrading ability were measured in order to include the enzymatic antioxidant component. Results: Short-term DR caused alterations in the total reducing power of various mouse tissues, indicating changes in the total scavenging ability of these tissues. Overall direct antioxidant capacity of heart, kidney and muscle was enhanced; liver and small intestine deteriorated; brain did not differ between DR and AL groups; lung and spleen exhibited a mixed response. Conclusions: We have shown for the first time that DR causes changes in the total reducing power of different mouse tissues, thus, affecting the overall direct antioxidant capacity. These findings support the suggestion that there may be a biological regulation of the antioxidant system.

Notes: Summary Background: The body's nucleotide pool derives from three potential sources: de novo synthesis, salvage of preformednucleosides/bases or the diet. The relative contributions of these pathways of assimilation are poorly understood in vivo. Dietary nucleotides have been suggested to have beneficial effects an the development and repair of the gastrointestinal tract. Tissues with a rapid turnover, such as the gut and the immune system cells, may utilise preformed nucleotides (coming from the diet), in situation in which there is a high demand of nucleotides for nucleic acid synthesis. Therefore, nucleotides could be onsidered as conditionally essential nutrients. Aim of the work and methods: -Development of a method to measure synthesis de novo of RNA-purine nucleotides in Caco-2 cells, relying an the incorporation of 14C-glycine into the purine ring of the nucleotide. To establish the fractional synthesis rate of RNA purine nucleotides in Caco-2 cells, grown in culture medium containing different concentrations of glutamine, in the presence or abscence of added nucleotides. To investigate the degree to which tissue ribonucleosides are derived from the culture medium or from de novo synthesis in the presence of different concentrations of glutamine, using undifferentiated Caco-2 cells, stressed or not by the addition of IL-1β to the medium. Results and conclusions:The presence of high levels of glutamine in the culture medium is essential for cell proliferation (estimated by measurement of the fractional synthesis rate of purine nucleotides) and the presence of nucleotides cannot replace the glutaminedependence of Caco-2 cell proliferation. The incorporation or exogenous purine nucleotides into RNA of Caco-2 cells is rather limited, and it becomes important when cells are stressed by glutamine deprivation. Stress by addition of interleukin-1β resulted in the maintenance or the increase in de novo synthesised RNA-purine nucleotides, even in the presence of exogenous nucleotides. However, the addition of interleukin-1β to the culture medium led to an enhanced salvage of preformed pyrimidine nucleotides for nucleic acid synthesis when glutamine was present in the medium at a concentration of 0.5 mmol/L.

Notes: Summary Scientific evidence is accumulating that meat itself is not a risk factor for Western lifestyle diseases such as cardiovascular disease, but rather the risk stems from the excessive fat and particularly saturated fat associated with the meat of modern domesticated animals. In our own studies, we have shown evidence that diets high in lean red meat can actually lower plasma cholesterol, contribute significantly to tissue omega-3 fatty acid and provide a good source of iron, zinc and vitamin B12. A study of human and pre-human diet history shows that for a period of at least 2 million years the human ancestral line had been consuming increasing quantities of meat. During that time, evolutionary selection was in action, adapting our genetic make up and hence our physiological features to a diet high in lean meat. This meat was wild game meat, low in total and saturated fat and relatively rich in polyunsaturated fatty acids (PUFA). The evidence presented in this review looks at various lines of study which indicate the reliance on meat intake as a major energy source by pre-agricultural humans. The distinct fields briefly reviewed include: fossil isotope studies, human gut morphology, human encephalisation and energy requirements, optimal foraging theory, insulin resistance and studies on hunter-gatherer societies. In conclusion, lean meat is a healthy and beneficial component of any well-balanced diet as long as it is fat trimmed and consumed as part of a varied diet.

Notes: Abstract Drift of mutated sectors in sectorial or mericlinal plant chimeras has been interpreted as indirect evidence of initial impermanence at the apex. However, the same effect may result from mutation in noninitial cells positioned close to the vertex of the apical dome. Clonal analysis of the cell packets present in the superficial layer of spruce and magnolia apices provided the library of patterns suggesting that the position and the number of initial cells, and in some cases also the meristem axis inclination, may change over time. Multicellular clones originating from a single cell have been found in the geometric center of some apices, whereas in other apices the cellular center (where three or four clonal borders meet) did not correspond to the geometric center of the apex. Such effects may result only from initial impermanence.

Notes: Abstract Temporal analyses of cell division and tissue expansion in pea, tobacco, and sunflower leaves reveal that both processes follow similar patterns during leaf development. Relative cell division and relative tissue expansion rates are maximal and constant during early leaf development, but they decline later. In contrast, relative cell expansion rate follows a bell-shaped curve during leaf growth. Cell division and tissue expansion have common responses to temperature, intercepted radiation, and water deficit. As a consequence, final leaf area and cell number remain highly correlated throughout a large range of environmental conditions for these different plant species, indicating that cell division and tissue expansion are co-ordinated during leaf development. This co-ordination between processes has long been explained by dependence between both processes. Most studies on dicotyledonous leaf development indicate that leaf expansion rate depends on the number of cells in the leaf. We tested this hypothesis with a large range of environmental conditions and different plant species. Accordingly, we found a strong correlation between both absolute leaf expansion rate and leaf cell number. However, we showed that this relationship is not necessarily causal because it can be simulated by the hypothesis of independence between cell division and tissue expansion according to Green's theory of growth (1976).

Notes: Abstract In turgid multicellular organs, it is convenient to differentiate between the two kinds of tensile forces acting in cell walls as a result of turgor pressure. The primary forces occur both in situ and in cells isolated from the organ, whereas the secondary forces occur only in situ. The latter are an unavoidable physical consequence of the variation in mechanical parameters of tissues forming layers or strands. The most rigid tissue is under maximal tensile force, whereas the least rigid is under maximal compressive force. These forces cause tissue stresses (that is, certain tissues are under tensile stress, whereas others are under compressive stress in the organ). The primary and secondary forces result in primary and secondary stress in cell walls, respectively. The anisotropy of the primary stress is a function of cell shape. For instance, in cylindric cells the anisotropy expressed as the ratio of longitudinal to transverse stresses is 0.5. The anisotropy of the secondary stress is a function of the compound structure of the organ. For example, in the epidermis of sunflower hypocotyl, the longitudinal secondary stress is much higher than the transverse stress. The primary and secondary stresses are superimposed, and, as a consequence, the stress anisotropy in the outer thick walls of epidermal cells is greater than 1. These outer epidermal walls transmit most of the tissue stress. When the epidermis is peeled but remains turgid, only primary stress remains, but loading of the peel can reestablish the original stress anisotropy. We studied the effect of stress anisotropy changes on the orientation of cortical microtubules (CMTs) in the sunflower hypocotyl epidermis. We showed that changes in stress anisotropy cause the CMT orientation to change in the direction of maximal wall stress. In situ, the relatively high tensile tissue stress in the epidermis causes maximal stress in the longitudinal direction and relatively steep CMT orientation. When the tissue stress is removed from the epidermis by peeling, the CMTs tend to reorient toward the transverse direction, which is the direction of maximal stress in the primary component. On application of external longitudinal stress, to substitute for tissue stress, CMTs tend to reorient in the longitudinal direction. However, a relatively high rate of plastic strain is caused by the stress applied to the peel in an acid medium. This produces a less steep orientation of CMTs. It appears that the change in stress anisotropy orients the CMT in the direction in which the stress is maximal after the change, but there is also some effect of the growth rate on the orientation.

Notes: Abstract In this article we describe a new method for the determination of turgor pressures in living plant cells. Based on the treatment of growing plant cells as thin-walled pressure vessels, we find that pressures can be accurately determined by observing and measuring the area of the contact patch formed when a spherical glass probe is lowered onto the cell surface with a known force. Within the limits we have described, we can show that the load (determined by precalibration of the device) divided by the projected area of the contact patch (determined by video microscopy) provides a direct, rapid, and accurate measure of the internal turgor pressure of the cell. We demonstrate, by parallel measurements with the pressure probe, that our method yields pressure data that are consistent with those from the pressure probe. Also, by incubating target tissues in stepped concentrations of mannitol to incrementally reduce the turgor pressure, we show that the pressures measured by tonometry accurately reflect the predicted changes from the osmotic potential of the bathing medium. The advantages of this new method over the pressure probe are considerable, however, in that we can move rapidly from cell to cell, taking measurements every 20 s. In addition, the nondestructive nature of the method means that we can return to the same cell repeatedly for periodic pressure measurements. The limitations of the method lie in the fact that it is suitable only for superficial cells that are directly accessible to the probe and to cells that are relatively thin walled and not heavily decorated with surface features. It is also not suitable for measuring pressures in flaccid cells.

Notes: Abstract A ``double-water-film electrode technique'' has been developed for the long-term characterization of the electrical properties across the interface between the nodal (N) and internodal (A or B) cells and the vacuole along the length of an internode of Chara as a function of time and temperature. The electrode unit consisted of a pair of the water-film electrodes described elsewhere (Chilcott 1988; Chilcott and others 1983; Coster and others 1984; Lucas 1985; and Ogata 1983). The distance between two water-film probes was fixed at 1.0 cm. By scanning the electrode unit, the spatial variations in electrical resistance and capacitance along the longitudinal axis of Chara were observed. Analysis was performed by applying an electrical equivalent circuit for the biomembrane (Philippson 1921). Across the internode (−A or −B)/central nodal cells interface, the specific parallel resistance (Rm) and the parallel capacitance (Cm) at 20°C were 30 ± 5 × 10−3Ωm2 and 1.5 ± 0.5 × 10−1Fm−2 (at 30 Hz), respectively. And the series resistance, corresponding to the vacuole of the internode was 8 × 10−3Ωm2. Study of temperature dependencies of Rm and Cm suggested that a dynamic homeostatic regulation was operating at the interface where numerous plasmodesmata were observed with an electron microscope (Pickett-Heaps 1967; Spanswick and Costerton 1967). Assuming that the individual cylinder of plasmodesma was filled only with cytoplasm, the number of plasmodesma per interface was estimated at 2.6 × 105.

Notes: Abstract The influence of putrescine (Put) and AgNO3 on shoot multiplication, in vitro flowering and endogenous titers of polyamines in Cichorium intybus L. cv. Lucknow local was investigated. Exogenous administration of Put at a concentration of 40 mM resulted in maximum tissue response in terms of shoot numbers (34.6 ± 2.61) and shoot lengths (7.6 ± 0.57 cm) on MS media supplemented with 2-iP (2.0 mg L−1) and GA3 (0.5 mg L−1) as observed on the 35th day. Exogenous application of 40 μM AgNO3 resulted in maximum shoot number (36.8 ± 2.63) and shoot lengths (7.9 ± 0.76 cm) on day 35 on the same media. Endogenous titers of conjugated spermidine decreased sharply from day 7–21, whereas endogenous conjugated spermine levels peaked on day 28 (1265 ± 94.9 nmoles g−1 FW), after treatment with 40 mM Put. Whereas, AgNO3 (40 μM) fed samples resulted in higher titers of endogenous conjugated spermine (1405 ± 105.6 nmoles g−1 FW, 3.62 fold over control) on day 14. All other treatments showed decreasing endogenous levels during the whole culture period. Both Put (40 mM) and AgNO3 (40 μM) resulted in floral initiation and floral development on day 28 and 14 (3.76 ± 0.16, 4.2 ± 0.21 flowers per shoot apices), respectively. To investigate the role of Put (40 mM) and AgNO3 (40 μM) on morphogenetic response and endogenous conjugated polyamine titers in shoots of chicory, polyamine inhibitors (DFMA and DFMO) were used. The morphogenetic response and the endogenous conjugated pool of polyamines were diminished in DFMA and DFMO treatments, but could be restored by addition of Put (40 mM) and AgNO3 (40 μM). Under exogenous Put feeding, ethylene production was reduced in shoot cultures of chicory. This study shows for the first time the influence of polyamines on multiple shoot initiation from axillary buds of C. intybus L. cv. Lucknow local and also indicates the promotive effect of Put and AgNO3 on autoregulation of polyamine biosynthesis, thereby regulating in vitro flowering, the endogenous pool of polyamines and shoot multiplication.

Notes: Abstract Plant responses to herbivores are complex. Genes activated on herbivore attack are strongly correlated with the mode of herbivore feeding and the degree of tissue damage at the feeding site. Phloem-feeding whiteflies and aphids that produce little injury to plant foliage are perceived as pathogens and activate the salicylic acid (SA)-dependent and jasmonic acid (JA)/ethylene-dependent signaling pathways. Differential expression of plant genes in response to closely related insect species suggest that some elicitors generated by phloem-feeding insects are species-specific and are dependent on the herbivore's developmental stage. Other elicitors for defense-gene activation are likely to be more ubiquitous. Analogies to the pathogen-incompatible reactions are found. Chewing insects such as caterpillars and beetles and cell-content feeders such as mites and thrips cause more extensive tissue damage and activate wound-signaling pathways. Herbivore feeding is not equivalent to mechanical wounding. Wound responses are a part of the induced responses that accompany herbivore feeding. Herbivores induce direct defenses that interfere with herbivore feeding, growth and development, fecundity, and fertility. In addition, herbivores induce an array of volatiles that creates an indirect mechanism of defense. Volatile blends provide specific cues to attract herbivore parasites and predators to infested plants. The nature of the elicitors for volatile production is discussed.

Notes: Abstract The term ``actinorhiza'' refers both to the filamentous bacteria Frankia, an actinomycete, and to the root location of nitrogen-fixing nodules. Actinorhizal plants are classified into four subclasses, eight families, and 25 genera comprising more than 220 species. Although ontogenically related to lateral roots, actinorhizal nodules are characterized by differentially expressed genes, supporting the idea of the uniqueness of this new organ. Two pathways for root infection have been described for compatible Frankia interactions: root hair infection or intercellular penetration. Molecular phylogeny groupings of host plants correlate with morphologic and anatomic features of actinorhizal nodules. Four clades of actinorhizal plants have been defined, whereas Frankia bacteria are classified into three major phylogenetic groups. Although the phylogenies of the symbionts are not fully congruent, a close relationship exists between plant and bacterial groups. A model for actinorhizal specificity is proposed that includes different levels or degrees of specificity of host-symbiont interactions, from fully compatible to incompatible. Intermediate, compatible, but delayed or limited interactions are also discussed. Actinorhizal plants undergo feedback regulation of symbiosis involving at least two different and consecutive signals that lead to a mechanism controlling root nodulation. These signals mediate the opening or closing of the window of susceptibility for infection and inhibit infection and nodule development in the growing root, independently of infection mechanism. The requirement for at least two molecular recognition steps in the development of actinorhizal symbioses is discussed.

Notes: Abstract Investigations were carried out during the period 1997-1999 on lepidopteran entomofauna deleterious to poplars (Populus spp.) in Bulgaria. The phytophages were collected in larval and pupal stage and were reared under laboratory con-ditions to the adult stage. In two regions of Bulgaria, Sofia and Montana, 18 new and little-known phytophages in seven families were established on the trees in Populus genus: Limenitis populi (L.) (Nymphalidae); Anacampsis populefia (Cl.), Gelechia turpella (Den. & Schiff.) (Gelechiidae); Synanthedon melliniformis (Lasp.) (Sesiidae); Euproctis similis (Fuessl.) (Lymantriidae); Drymonia velitaris (Hufn.), Notodonta tritophus (Den. & Schiff.) (Notodontidae); Amphipyra pyramidea (L.), Catocala nupta (L.), Cosmia trapezina (L.), Enargia paleacea (Esp.), Eupsilia transversa (Hufn.), Ipomorpha subtusa (Den. & Schiff.), Parastichtis ypsilon (Den. & Schiff.), Scoliopteryx libatrix (L.) (Noctuidae); Gastropacha quercifolia (L.), Phyllodesma tremulifolia (Hb.) and Poecilocampa populi (L.) (Lasiocampidae). G. turpella and S. melliniformis are new records for Bulgaria and D. velitaris was found for the first time in Northern Bulgaria. Thirteen species (L. populi, E. similis, D. velltaris, N. tritophus, A. pyramidea, C. nupta, C. trapezina, E. transversa, I. subtusa, P. ypsilon, G. quercifolia, P. tremulifolia and P. populi) were established for the first time in Bulgaria to have a trophic relationship with poplars. G. turpella and L subtusa were occurred in the highest population densities – up to 1.6 larvae/m and up to 1.1 larvae/m respectively. The remaining species occurred in low population densities in the studied habitats and were not considered to be pests on the poplars.

Notes: Abstract The commercial available diatomaceous earth Fossil Shield® and the neem product Neem Azal-T/S® are tested as single treatments and in combination as controls for the stored product pests Sitophilus oryzae (L.) and Tribolium castaneum (Herbst). The diatomaceous earth, applied in concentrations of 0.5 g, 1 g and 2 g kg-1 rice, reduced numbers of surviving beetles significantly (over a period of 3 weeks). A single treatment with the neem product, in concentrations of 0.01 g, 0.1 g, 0.2 g and 1 g azadirachtin kg-1 rice, increased the mortality rate for both species significantly. The combination of neem and diatomaceous earth (1 g diatomaceous earth with 0.2 g or 1 g azadirachtin kg-1 rice) was more effective than the single treatment in reducing numbers of surviving beetles.

Notes: Summary The Russian wheat aphid (=RWA), Diuraphis noxia (Kurdj.) is an exotic expansive aphid on the small grains determined in the Czech republic since 1993 (1995). Determination of RWA flight patterns by suction air traps (= SATs) between 1994-1998 indicated some banker, less important, or indicated RWA distribution areas. A comparison of the RWA monitoring by SATs and by crop samples in areas more or less distant from individual SAT locations manifested a high silmilarity of RWA evidence in its overall distribution and area-influenced population numbers. Owing to geographic area-dependent features of its life-cycle (holocycly) and host plant preferences, RWA seems to be most injurious to spring barley. In spite of the role of SATs in aphid forecasting, RWA samples are recommended to be taken also from the crops at the early ripening stage as the respective SAT evidence reflects only the emigration of the alate aphids originating from the decreasing populations. In general, RWA detection by SATs was either simultaneously or successively found to reflect the RWA presence in the more or less distant crops. The 30km perimeter of a SAT is agreed to be representative for mon-toring RWA populations in a sampled area.

Notes: Abstract A supplementary research on Russian wheat aphid (= RVA), Diuraphis noxia (Kurdj.) between 1998-99 documented the pest distribution all over the Czech republic already. RVA abundance was proved to be significantly area-dependent within the state. The banker areas are in southern Moravia (district Znojmo, the frontier area near Austria) up to central Moravia. The determined RWA distribution area is much broader than presumed on grounds of climatic characteristics. The abundance of RVA generally decreases from the SE to the NW and N direction, as well as with the increasing altitude within the area.The highest altitude a.s.l. of RWA occurrence in the crop was 762 in. The re-classified main expansion route was determined to follow two directions (sourcing from Hungary and Austria) to southern-central-northem Moravia, and to southern Bohemia and further on to the central Bohemian lowland. The field information indicated the development of preference of RWA for barley.

Notes: Abstract Aiming to control the citrus leafminer Phyllocnistis citrella St. while at the same time protecting the biological equilibrium and mainly the beneficial insect Cales noacki, experiments have taken place in the regions of Argolis and Fthiotis, Greece in 1996 and 1997. Insegar (fenoxycarb) 2 5 WP was applied on young citrus trees. During 1996, the compound, applied every 10 days at a rate of 30 g/hI (with or without adjuvant), gave satisfactory protection from P. citrella without substantially harming C. noacki populations. No phytotoxicity symptoms were observed during the experiments.

Notes: Abstract This study demonstrates a laboratory induction of a malathion - resistant strain in Amblyseius cydnodactylon Shehata & Zaher and effect of selection on reproduction. Initially 500 sensitive females obtained from a laboratory mass culture were exposed to malathion at LC70. Subsequent selections were conducted every two generations at progressive LC70 values and number of eggs/female/day was recorded at each selection. Experiments were carried out under laboratory conditions of 24-28°C and 70-80 % R. H. The LC70 in the parent generation was 5.19 ppm and increased to 20 ppm in F4 selection generation. The dosage mortality relationships continued to increase up to a maximum of 282.3 ppm in F16. The rate of developing resistance increased from 1.75 folds in F2 to 1.97 folds in F4 and gradually reached a maximum of 54.39 folds in F16. There was an obvious decrease in reproduction corresponded to increasing resistance. For example, the number of eggs/female/day in F 16 was 8 eggs, contrasting 3 eggs in the parent generation.

Notes: Abstract A survey on the present distribution of the horse chestnut leafmining moth, Cameraria ohridella Desch. & Dimic 1986, in Europe and in Italy is given: In Europe, within a period of 15 years, the leafminer – recently introduced into Central Europe – has settled in an area that spreads over 13 latitudinal and about 20 longitudinal degrees; in Northern Italy, an area of approximately 50,000 km2 has been afflicted in the 5 years since the leafminer's introduction in South Tyrol and Julian Venetia in 1993. The development of the distribution in the region of South Tyrol and Trentino is shown: In the province of Bozen-South Tyrol, between 1995 and 1999 the moth has spread in all the three principal valleys (Eisacktal, Pustertal, Etschtal); in the Trentino province, first local attacks began only in 1998, near Trento and Riva, but spread over the entire Etschtal/Adige Valley in 1999. While in South Tyrol C. ohridella was introduced from the North (North Tyrol), the introduction into the Trentino occured from the South (Verona). At present the attacks in Trentino province are preponderantly low (resulting heavy only in the city of Trento and in Riva del Garda) and limited to altitudes ranging from 70 to 700 in. In South Tyrol, attacks are mainly heavy in altitudes ranging from 250 to 900/950 m, while they are low in the increasingly affected higher altitudes of 1,000 to 1,230 m. Numeric attack parameters are given to show attack intensities. The number of generations per year depends on the respective climatic conditions: in lower-altitudes, usually three generations develop in a year, but in higher altitudes (800 to 1,100 m) only two generations; in the climatically favoured area around the Lake Garda a (partial) fourth generation appears possible. The question of host plants (Aesculus sp., Acer sp.) and the larval parasitism of C. ohridella is discussed. The parasitism of the larvae, principally by Eulophidae (Hym., Chalcidoidea), was ascertained already in the first years of attack: In South Tyrol, 16 species of parasitoids appeared, and 8 in the Trentino province, but only two species were dominant: Minotetrastichus sp. and Pnigalio sp. The level of parasitism reached was low, comparable to other Central European countries.

Notes: Abstract The reproductive potential of Microplitis rufiventris Kok. females resulted from Spodoptera littoralis (Boisd.) larvae topically treated by a constant dose (70 pg/5 pl) of the growth regulator precocene II at different intervals of post-parasitism was studied in the laboratory. The obtained results showed that the effects of PH vary according to the timing of treatment relative to larval development of the parasitoid. Reduction in reproductive capacity of the parasitoid occurred when PII was applied on host larvae containing young parasitoid larvae. Workers should be aware from the delayed effects of precocenes which may occur in later stages of the non-target insects.

Notes: Abstract The effects of the insecticide lambda-cyhalothrin (Karate) on non-target arthropods in winter wheat were studied throughout two successive seasons in 1998 and 1999. The study particularly focussed on the crop in the growing season after insecticide application (also winter wheat) for detection of potential long-term effects and for determination of the suitability of different sampling methods.The investigations were based on the assumption that arthropod immigration from surrounding areas is limited in large fields. For this reason a simple approach seemed to be feasible. Two plots of equal size (10 ha, adjusted to each other) were defined in a 100 ha field and designated control and treatment plots. Ten sampling points were established on each plot. The following monitoring methods were utilised: visual counting, sweep netting and pitfall trapping. In the first year of investigation, countings and catches were carried out 2 days prior to insecticide application and 2, 16, 30 and 44 days after application. In the next year, they were carried out 365 days and 384 days after insecticide application in the successive crop of winter wheat.At the time of the first sampling prior to insecticide application, the two plots showed significant differences with respect to arthropod density or activity, particularly in visual counting and to a minor degree to sweep netting and pitfall trapping. Lower densities or activities were observed in the plot reserved for treatment. Measures for mathematical equalisation of the results of population densities before pesticide treatment should be considered.After insecticide application, the densities or activities of non-target arthropods decreased, particularly in visual counting and sweep netting. After one year, these effects disappeared to a large extent. Several groups of arthropods reached even higher levels in the treated plot than in the untreated one. The pitfall traps revealed weak activity-decreasing effects in carabids and spiders in the treated plot, but the opposite tendency for staphylinids. Hence, it seems that the conditions in a large field are less homogeneous, and that smaller scale conditions can support processes of recovery in non-target populations.

Notes: Abstract The history of the controversy overrecombinant bovine growth hormone (rBGH) is exploredin terms of the issue of the potential robustness ofa consumption-driven ``new'' politics of food andagriculture. It is noted that while the dominanthistorical traditions in the social sciences haveserved to discount the autonomous role that consumersand consumption play in modern societies, there hasbeen growing interest in consumption within foodstudies as well as other bodies of scholarship such aspostmodernism, social constructivism, socialcapital/social distinction, and environmentalsociology. A review of the shifting pattern ofdiscourses during the rBGH controversy shows thatconsumption-driven claims and politics played atangible, but relatively minor role. Even so, it issuggested that the rBGH experience along with paralleltrends in food politics (e.g., anti-pesticidecampaigns such as the ``Alar scare,'' agribusinessattempts to intimidate opponents through fooddisparagement laws, conditions-of-productionprovisions of the World Trade Organization agreement)could make the consumption/consumer dimension of foodpolitics more important in the future.

Notes: Abstract The ten largest food and beveragecorporations control over half of the food sales inthe United States and their share may be increasing.Using data from a range of secondary sources, weexamine these corporations and their boards ofdirectors. Social and demographic characteristics ofboard members gleaned from corporate reports, thebusiness press, and elsewhere are presented.Information on interlocking corporate directorates andother common ties among members of the boards ofdirectors show that US based food and beveragecorporations are tied together through a web ofindirect interlocks.

Notes: Abstract This introduction situates key themesfound in papers given at a recent workshop on thechanging material practices, meanings, and regulationof US organic food production. The context is theemergence of an international bio-politics ofagriculture and food and, more particularly in the US,the contradictions of sustainable agriculturemovements catalyzed by the rapid scaling up of organicagriculture from a niche activity to nascentindustry.

Notes: Abstract As the organic food sector has grownand changed to become more mainstream, large-scaleconventional growers have entered into organicproduction. While it is increasingly clear that notall organic farms are self-sufficient small scaleunits that practice poly-cultural agronomy and sell inlocal marketing venues, there still exists apresumption that there are clear lines between thesmall scale ``movement'' farmers who followagro-ecological agronomic ideals and the relativelylarger and partly conventional newcomers who do not.This paper addresses a specific empirical issue, whichis the extent to which California organic farmerspractice the techniques of ecological farming. Itillustrates that while there are some distinctdifferences in practices between larger and/orpart-conventional (i.e., mixed) growers and smallerand/or all-organic growers, it also shows that inalmost all cases, practices fall quite short ofagro-ecological ideals. By examining in more depth howgrowers follow particular agro-ecological principles,the paper also demonstrates that key variations arerelated to variables separate from scale. Some ofthese variables are geographic, ranging frombiophysical and climatic opportunities andconstraints, to regional norms and institutionalsupport. Mostly, however, variation is related to cropspecificities and the availability of efficacioustechnologies to deal with crop-specific problems. Thisso-called technology barrier crucially depends on howorganic is defined, and thus suggests the importanceof organic rules and regulations in shaping thepractices of organic production.

Notes: Abstract Actor network theory and supply chainmanagement theory provide suggestive researchdirections for understanding regional agri-foodnetworks. These theories claim that relationshipsbased upon trust and cooperation are critical to thestrength and vitality of the network. This means thatexploring and detailing these relationships among thesuppliers, producers, workers, processors, brokers,wholesalers, and retailers within specific regionalgeographies of these networks are critical forfurthering cooperation and trust. Key areas ofcooperation include resource sharing andapprenticeship programs. Employing food networks as akey unit of contextual analysis will deepen ourunderstanding of how to enhance their resiliency andvibrancy. Important questions can be raised about thedifference gender makes for farmers, brokers,entrepreneurs, and workers in local food networks.

Notes: Abstract The international organic agricultureand fair trade movements represent importantchallenges to the ecologically and sociallydestructive relations that characterize the globalagro-food system. Both movements critique conventionalagricultural production and consumption patterns andseek to create a more sustainable world agro-foodsystem. The international organic movement focuses onre-embedding crop and livestock production in ``naturalprocesses,'' encouraging trade in agriculturalcommodities produced under certified organicconditions and processed goods derived from thesecommodities. For its part, the fair trade movementfosters the re-embedding of international commodityproduction and distribution in ``equitable socialrelations,'' developing a more stable and advantageoussystem of trade for agricultural and non-agriculturalgoods produced under favorable social andenvironmental conditions. The international market forboth organic and fair trade products has grownimpressively in recent years. Yet the success of thesemovements is perhaps better judged by their ability tochallenge the abstract capitalist relations that fuelexploitation in the global agro-food system. While theorganic movement currently goes further in revealingthe ecological conditions of production and the fairtrade movement goes further in revealing the socialconditions of production, there are signs that the twomovements are forging a common ground in definingminimum social and environmental requirements. I arguefrom a theoretical and empirical basis that what makesfair trade a more effective oppositional movement isits focus on the relations of agro-food trade anddistribution. By demystifying global relations ofexchange and challenging market competitiveness basedsolely on price, the fair trade movement creates aprogressive opening for bridging the wideningNorth/South divide and for wresting control of theagro-food system away from oligopolistic transnationalcorporations infamous for their socially andenvironmentally destructive business practices.

Notes: Abstract Values and beliefs regarding communityfood security were investigated among participants in2–3 day participatory planning events related to thelocal food system in six rural counties from oneregion of upstate New York. The results of Qmethodology reveal three distinct viewpoints: a) theSocial Justice viewpoint, which is primarily concernedwith hunger and the potential harm caused by welfarereform; b) the Pragmatist viewpoint, which values thecontributions agriculture makes to local communitiesand is not concerned about environmental or socialexternalities of the dominant food system; and c) theVisionary viewpoint, which also values agriculture inthe community but is very concerned aboutenvironmental and social externalities. After theplanning events, the Pragmatist viewpoint experiencedan 88% increase in members and the other twoviewpoints became less salient. Various categories ofprofessionals (e.g., nutrition, social welfare,agriculture, environmental) tend to express theviewpoints associated with their professions and/orthe client groups they serve. Despite thesedifferences among participants, the planning events inall six counties resulted in a wide range of goals andobjectives centered on a theme of re-localizing avariety of food system activities. These results arediscussed in relation to the desirability ofdeveloping an explicit philosophy of food andagriculture and the ideal processes required to doso.

Notes: Abstract Various structural factors influenceorganic farmer decision-making. Analyses that combinestructure and agency provide an opportunity forunderstanding farmers' perceptions of the political,economic, and social ``world'' in which they operate.Rich conversational interviews, conducted with twentycertified organic farmers in Illinois and analyzedwith multiple qualitative methods, show how farmersmediate structural concerns. In addition to political,economic, and social structures, a fourth structure isneeded. Indeed these organic farmers emphasize theimportance of ecological factors in theirdecision-making. Within the perceived economic,political, social, and ecological structures, numeroustopics (i.e., marketing, policy, family, ecosystems)and subtopics (i.e., diversification, farm programs,traditions, soils) exist. Farmers' quotations providedetailed information of how they view and mediatestructures in their daily on-farm decision-making.

Notes: Abstract In the developmentalist era,industrialization has simultaneously transformedagriculture and degraded its natural and culturalbase. Food production and consumption embodies thecontradictory aspects of this transformation. Thispaper argues that the crisis of development hasgenerated two basic responses: (1) the attempt toredefine development as a global project, includingharnessing biotechnology to resolve the food securityquestion, and (2) a series of countermovementsattempting to simultaneously reassert the value oflocal, organic foods, and challenge the attempt on thepart of food corporations and national and globalinstitutions to subject the food question to marketsolutions. It is proposed that the power of food liesin its material and symbolic functions of linkingnature, human survival, health, culture and livelihoodas a focus of resistance to corporate takeover of lifeitself.

Notes: Abstract Colonists, unlike indigenous peoples, are often assumed tohave little knowledge of their environment. However, their perceptions of the environment and their knowledgeof natural resource systems have a significant impact on their farming practices. Farmers in the frontier regionof Marabá, Eastern Amazonia, understand nutrient cycling and the links between different components in farmingsystems. Diagrams drawn by farmers show very diversified systems, and farmers' knowledge of soilcharacteristics, including sub-surface features, and distribution in their localities is very detailed in comparison to pedologicalclassifications. However, knowledge about nutrient cycling is very uneven, even between farmers from the same area.Generally, farmers were found to have very detailed knowledge of environmental resources, but very patchyknowledge of processes and functions underlying systems, and this conforms to evolutionary models of ecologicalknowledge. Perceptions of change in soil fertility are related to the length of settlement, and are closelylinked to the presence of forest. Overall, the majority of farmers believe they will not be able to sustain cropping in thefuture, and as forest and fallow become scarce the most feasible option will be for them to move to other areas.Farmers are more optimistic about pasture, which is viewed as a more stable system, with the key to long-termsustainability being weed control. These findings imply that a high degree of information sharing between farmers andscientists is required to establish resource management strategies and social institutions to supportsustainable development strategies at the frontier.

Notes: Abstract Until recently, ethics was a highlyabstruse activity, with little reference to everydayaffairs. It dealt largely with what is calledmetaethics, and was in danger of becoming moribund asan intellectual activity. But for some years,ethics has been undergoing a process of rejuvenationand development. We now seem to be experiencing thebirth of this new discipline (or at least in the EU –the US has been engaged in it somewhat longer). The EurSafeCongress held at Wageningen University, March 4–6,1999 exemplifies this rejuvenation, and itstrongly suggests that a new discipline is emerging, that is not only exciting from an intellectualperspective, but also addresses issues of fundamentalsocial and political concern. It can beargued that, in this context, ethicists are in the position of guides.It is not their job to pronounce on what is right andwrong, but having trodden many of the theoreticalpaths through the forest, they are in a position toadvise and facilitate sound ethical decision-making byothers. The need for ethical insight in this field islikely to progressively increase over the comingyears. Ethicists have a duty to respond to this need.

Notes: Abstract Public concern over environmentalquality and food safety has culminated in thedevelopment of markets for “green” foods – foodsthat are variously construed as fresh, chemical-free,nutritious, natural, or produced in anenvironmentally-sustainable manner. Understanding theemergence of “green” foods is dependent on analysisboth of the ways in which foods are produced andprocessed, and of the meanings that are attached tothem at each stage of their production,transformation, and consumption. The notion of “green”foods is thereby understood here as a fluid andcontestable signifier that myriad actors involved inthe production/consumption cycle may attempt to shapefor their own purposes. This paper explores corporate capital's recent attempts, through certification logosand advertising, to signify the “healthiness” andenvironmental virtues of organically-produced foods inAustralia and New Zealand. These attempts have not,however, been universally successful either in termsof gaining consumer interest, or in gaining agreementsbetween farmers, certifying organizations, andcapitalist firms over the meaning of “organic” and thepractice of “sustainable” agriculture. The experienceof corporate involvement in the organics industry isillustrative of yet-to-be-resolved processes ofreflexive modernization. As food production andtransformation continues to produce environmental andsocial risks, the question of just what makes food“green” will continue to be a source of social conflict.

Notes: Abstract The performances in the food and cashcrop sectors and the availability and consumption ofagricultural inputs in Tanzania during structuraladjustment programs (1986–1996) are compared withperiods prior to this IMF/World Bank backed reform.The positive developments in the first five years ofreform appear to be not sustainable. Presentlyproductivity levels per rural capita for importantfood and cash crops are declining. There are nofurther improvements in the availability andconsumption of agricultural inputs. The removal ofsubsidies on agricultural inputs from 1991 onwards iscrucial in explaining the decline in maize production,the main food crop in Tanzania. Some assumptionsbehind the reform measures are proving to be wrong.Modifications are thus needed to improve theagricultural sector of Tanzania in a sustainable way.Structural adjustment programs usually go too far inreducing the role of the government. However,government involvement might be necessary to ensure ahigher consumption of agricultural inputs and thus abetter performance of the agricultural sector inTanzania. A higher productivity in the agriculturalsector is needed in Tanzania to avoid food securityproblems in the future.

Notes: Abstract Since its inception in 1988, the SAREprogram has sponsored hundreds of projects to exploreand apply economically viable, environmentally sound,and socially acceptable farming systems. Recognizingthat researchers often collaborated with producers andthat producer interest in sustainable agriculture wasincreasing, SARE's North-Central Region began directlyfunding farmers and ranchers in 1992 to test their ownideas on sustainable agriculture. The present articleis based on data from the formative evaluation of thefirst five years (1992 to 1996) of the NCR-SAREProducer Grant Program. The evaluation used acombination of mail surveys, non-response telephoneinterviews, and personal interviews.The evaluation revealed that the Program hassucceeded in showing that sustainable agriculturaltechnologies and practices can be viable andprofitable alternatives to conventional ways ofproducing crops and animals while simultaneously beingless environmentally damaging. On the other hand, thecontributions of the Producer Grant Program to thesocial and institutional spheres in which agricultureis embedded are less clear. Changes in these spheresare imperative for the success of sustainableagriculture, and for it to become more mainstream.Such changes cannot occur overnight, but they willremain the main challenge for SARE to tackle in thenear future.

Notes: Abstract Adoption of integrated pest management(IPM) practices in the Guatemalan highlands has beenlimited by the failure of researchers andextensionists to promote genuine farmer participationin their efforts. Some attempts have been made toredress this failure in the diffusion-adoptionprocess, but farmers are still largely excluded fromthe research process. Understanding farmers'agricultural knowledge must be an early step toward amore participatory research process. With this inmind, we conducted a semi-structured survey of 75Cakchiquel Maya farmers in Patzún, Guatemala, tobegin documenting their pest control practices. Theirresponses revealed that their understanding ofbiological and curative pest control is limited.However, their broad knowledge of cultural preventivepest control practices could explain why they hadfaced few pest problems in their traditionalmilpa (intercrop of corn, beans, and other edibleplants). The majority of these preventive practicesare probably efficient and environmentallyinnocuous.

Notes: Abstract Global competition has made thetraditional sugarcane and pineapple industriesincreasingly non-viable in Hawaii. One initiative torevive the agricultural sector calls for diversifyinginto non-traditional export crops that gains highervalue by attaching the paradise identity such as freshpineapples, macadamia nuts, or tropical flowers.Drawing from cases of pineapples and macadamia nuts,this paper examines how Hawaii's foodstuffs were ableto capture a premium value of place-association due tothe social construction of Hawaii as a place. Anexpansion of the niche markets, however, has allowedthe symbolic meaning of these products to beappropriated and reconstituted by global interests.Without confronting the fundamental problemsassociated with land and labor relations in Hawaii,Hawaii producers are caught in a niche-markettreadmill. They continuously seek new potentials forhigh-value crops, but are unable to maintain controlbeyond their inventive stage.

Notes: Abstract The Forestry Pilot Plan set intomotion collectively-owned and managed forestry in overforty communities in Quintana Roo, Mexico and hasshown the promise of a forestry development model thatpromotes conservation by giving local people a genuinestake in sustainable resource management. Today, thelegacy of the PPF is under great pressure. Externally,neoliberal policy reform restructures agrarianproduction in ways that favor individual overcollective management of natural resources.Internally, organizational problems createinefficiencies within both forestry ejidos(cooperative agrarian communities) and theirintermediate level forestry civil societies. Peasants'capacity to defend their interests and dealeffectively with their production problems throughstrong representative organizations is beingundermined by new rules for economic associationwithin the ejidos and by the turning over of technicalservice financing to the market. Though organizationalinnovations within the ejidos hold positive potential,existing civil societies merit continued assistance askey actors promoting sustainable forestry. Studyingcommon property management regimes across multiplelevels and dimensions reveals that in Mexico, policyreform overlooks the crucial social resourcesrepresented by peasant organization, undermining thepossibility of sustainable forest management whileassigning the peasant most of the cost ofconservation. If conservation is indeed encouraged bythe genuine participation of those with a stake insustainable use of natural resources, national andinternational communities that value Mexico's tropicalforests should also invest in both social and economiccosts of conservation.

Notes: Abstract The proposed federal regulation oforganic agriculture in the United States raisesquestions both about the nature and character oforganic farming, as well as its relation to theagro-food system at large. The regulatory process hasengendered a public debate about conventional andalternative approaches to agricultural production,which in turn raises issues of environmental politicsand society-nature relations. An analysis oftranscripts from public hearings, organic farmingmovement literature, and interviews with organicpractitioners and advocates reveals the broaderecological, social, and political ramifications. Inexamining the proposed federal rule and its criticalopposition, we encounter two different worldviews; twoconflicting visions of agriculture, rural life, andnature itself. Whether this is a fundamental impasse,or a controversy that can be fruitfully resolved,remains an open question. But organic farming hasarrived at a critical juncture, both fraught withperil, and full of opportunity. The enormous publicresponse to this issue indicates the renewed potentialon the part of civil society to participate ingrassroots environmental social movements in supportof alternative agriculture.

Notes: Abstract Sustainable agriculture has lately madesignificant inroads into US agricultural policydiscourse. An examination of the ``life cycle'' of theFund for Rural America, a component of the 1996 farmbill, provides an example of the complex and contestedways in which the goals of sustainable agriculture areadvocated, negotiated, and implemented at the level ofnational policy, in the context of the evolvingpolitical and institutional arrangements of Americanagricultural policy. The Fund, with its relativelylarge endowment of $100 million annually, and itsexplicit emphasis on alternative agriculture research,is emblematic of both the growing politicaleffectiveness of the alternative agriculture movementand the increasing institutionalization of alternativeagriculture representatives in Federal agencies. Theuntimely demise of the Fund in the appropriationsprocess, however, illustrates the extent to whichcertain key spaces within the state remain outsidesustainable agriculture's broadening sphere ofinfluence. This suggests that while some aspects ofthe movement's organizing strategy are indeedeffective, some may need to be rethought in light ofthe experience with the FRA.