Effects of elevated CO2 and transgenic Bt cotton on plant chemistry, performance, and feeding of an insect herbivore, the cotton bollworm

Effects of elevated atmospheric CO₂ (double‐ambient CO₂) on the growth and metabolism of cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), fed on transgenic Bacillus thuringiensis (Berliner) (Bt) cotton [Cry1A(c)], grown in open‐top chambers, were studied. Two levels of CO₂ (ambient and double‐ambient) and two cotton cultivars (non‐transgenic Simian‐3 and transgenic GK‐12) were deployed in a completely randomized design with four treatment combinations, and the cotton bollworm was reared on each treatment simultaneously. Plants of both cotton cultivars had lower nitrogen and higher total non‐structural carbohydrates (TNC), TNC:Nitrogen ratio, condensed tannin, and gossypol under elevated CO₂. Elevated CO₂ further resulted in a significant decrease in Bt toxin level in GK‐12. The changes in chemical components in the host plants due to increased CO₂ significantly affected the growth parameters of H. armigera. Both transgenic Bt cotton and elevated CO₂ resulted in a reduced body mass, lower fecundity, decreased relative growth rate (RGR), and decreased mean relative growth rate in the bollworms. Larval life‐span was significantly longer for H. armigera fed transgenic Bt cotton. Significantly reduced larval, pupal, and adult moth weights were observed in the bollworms fed elevated CO₂‐grown transgenic Bt cotton compared with those of bollworms reared on non‐transgenic cotton, regardless of the CO₂ level. The efficiency of conversion of ingested food and of digested food of the bollworm were significantly reduced when fed transgenic Bt cotton, but there was no significant CO₂ or CO₂× cotton cultivar interaction. Approximate digestibility of larvae reared on transgenic cotton grown in elevated CO₂ was higher compared to that of larvae fed non‐transgenic cotton grown at ambient CO₂. The damage inflicted by cotton bollworm on cotton, regardless of the presence or absence of insecticidal genes, is predicted to be more serious under elevated CO₂ conditions because of individual compensatory feeding on host plants caused by nitrogen deficiency.     

Fitness of Bt‐resistant cabbage loopers on Bt cotton plants

Development of resistance to the insecticidal toxins from Bacillus thuringiensis (Bt) in insects is the major threat to the continued success of transgenic Bt crops in agriculture. The fitness of Bt‐resistant insects on Bt and non‐Bt plants is a key parameter that determines the development of Bt resistance in insect populations. In this study, a comprehensive analysis of the fitness of Bt‐resistant Trichoplusia ni strains on Bt cotton leaves was conducted. The Bt‐resistant T. ni strains carried two genetically independent mechanisms of resistance to Bt toxins Cry1Ac and Cry2Ab. The effects of the two resistance mechanisms, individually and in combination, on the fitness of the T. ni strains on conventional non‐Bt cotton and on transgenic Bt cotton leaves expressing a single‐toxin Cry1Ac (Bollgard I) or two Bt toxins Cry1Ac and Cry2Ab (Bollgard II) were examined. The presence of Bt toxins in plants reduced the fitness of resistant insects, indicated by decreased net reproductive rate (R₀) and intrinsic rate of increase (r). The reduction in fitness in resistant T. ni on Bollgard II leaves was greater than that on Bollgard I leaves. A 12.4‐day asynchrony of adult emergence between the susceptible T. ni grown on non‐Bt cotton leaves and the dual‐toxin‐resistant T. ni on Bollgard II leaves was observed. Therefore, multitoxin Bt plants not only reduce the probability for T. ni to develop resistance but also strongly reduce the fitness of resistant insects feeding on the plants.     

Development of <Emphasis Type="Italic">Cleruchoides noackae</Emphasis>, an egg-parasitoid of <Emphasis Type="Italic">Thaumastocoris peregrinus</Emphasis>, in eggs laid on different substrates,with different ages and post-cold storage

Cleruchoides noackae Lin and Huber (Hymenoptera: Mymaridae) is an egg parasitoid of Thaumastocoris peregrinus Carpintero and Dellapé (Hemiptera: Thaumastocoridae). The parasitism and development of C. noackae was studied in T. peregrinus eggs of different ages, laid on eucalyptus leaves or paper towel and stored at 5 °C. The emergence, sex ratio and development of C. noackae and hatched nymphs of T. peregrinus were evaluated. This parasitoid had an emergence rate higher than 60% from zero to one, one to two, and two to three-day old eggs and lower than 10% for those 3–4 and 4–5 days old. The female proportion was 78% and the egg-adult period for C. noackae was 19.5 days. The use of T. peregrinus eggs up to three days old, laid on paper towel and stored at 5 °C for 14 days did not affect the biological parameters of C. nockae and should be used for mass rearing of this parasitoid.     

Addition of a surfactant to tryptic soy broth allows growth of a lactic acid bacteria food antimicrobial, Escherichia coli O157:H7, and Salmonella enterica

Aims: This study aimed to determine the survival and growth of Escherichia coli O157:H7 and Salmonella enterica subsp. enterica in a medium supporting the growth of a Lactic Acid Bacteria (LAB) food antimicrobial culture. Methods and Results: Foodborne pathogens and LAB were cultured individually in tryptic soy broth (TSB), tryptic soy broth supplemented with one g l^?1 Tween 80^® (TSB‐T80), and de Man, Rogosa and Sharpe (MRS) broth. Growth of E. coli O157:H7 and Salmonella was similar in TSB and TSB‐T80 but was significantly less in MRS. Conversely, LAB growth was similar in MRS and TSB‐T80 but was significantly less in TSB. Conclusions: Supplementation of TSB with Tween 80^® allows growth of LAB to levels similar to that observed with MRS but does not inhibit the growth of E. coli O157:H7 and Salmonella. We present the formulation of a medium useful in studies useful for evaluating competitive inhibition of foodborne pathogens by LAB in vitro. Significance and Impact of the Study: This study reports the utility of TSB‐T80 for the completion of in vitro competitive inhibition assays incorporating a Lactic Acid Bacteria food safety culture.     

The combined effect of soil‐applied azadirachtin with entomopathogens for integrated management of western flower thrips

Performance of soil application of azadirachtin products with entomopathogens was evaluated as single treatments, in multiple combinations and in different concentrations against the soil stages of western flower thrips, Frankliniella occidentalis (Pergande), in French beans Phaseolus vulgaris L. Treatments consisted of NeemAzal‐T solution, neem pellets, Steinernema carpocapsae (Weiser) Nemastar^®, isolates of Metarhizium anisopliae (Metschnikoff) Sorokin (IPP 2539 and ICIPE‐69) and Beauveria bassiana (Balsamo) – Naturalis^®. All treatments were analysed for the number of emerging adults, while emerged adults in fungi‐based treatments were analysed additionally for the retarded development of mycosis as a possible cause of secondary mortality. Possible interactive effects in combined treatments were analysed using a generalized linear model (GLM) approach, and three levels of dose–response combinations of the selected treatments were further tested. Bioassay results of the single treatments indicated between 43% and 60% reduction in adult emergence with NeemAzal‐T solution thus proving to be the most efficient. However, most cadavers with entomopathogenic fungi (EPF) treatments showed the development of mycosis. Therefore, the reduction in adult emergence attributed to the EPF was altogether >87%. Combined treatments with Steinernema, Metarhizium (ICIPE‐69), NeemAzal‐T and neem pellets resulted in total reduction in adult emergence of 95–97% when late mortality by mycosis was considered. Of the treatment combinations, two showed synergistic, four additive and one an antagonistic response. Combining low concentration of entomopathogenic nematode (EPN) (100 IJ/cm²) with NeemAzal‐T resulted in satisfactory control compared to the operational dose of EPN, while the highest concentration of M. anisopliae (10⁸ conidia) combined with Steinernema showed the best performance with 74% reduction in adult emergence.     

The potential global distribution of the Bronze bug Thaumastocoris peregrinus Carpintero and Dellapé (Hemiptera: Thaumastocoridae)

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Synthesis of cinnamyl acetate catalysed by highly reusable cotton-immobilized Pseudomonas fluorescens lipase

Cinnamyl acetate as an important fragrance ingredient could be synthesized by lipase-catalysed transesterification in organic systems, but enzyme proteins tended to denature and inactivate for no water lubrication. To improve the non-aqueous stability of lipases, absorbent cotton was taken as an alternative “water” phase to stabilize enzyme proteins. In a mass ratio of 1:1, Pseudomonas fluorescens lipase was immobilized on cotton fibres by physical absorption in a column glass bottle, forming a facile cotton-lipase bioreactor in which the transesterification between cinnamyl alcohol and vinyl acetate processed efficiently. From the molar conversions after reaction for 2?h at 37?°C and 160?rpm, the ability of cotton-lipase to transform substrate was more than 5-folds of native lipase. And even in static state and at 4?°C, the conversion of reaction catalysed by cotton-PFL had 11-fold increase relative to native lipase after 8?h. Recycles showed that the cotton-lipase had an extra-long half-life of activity (t_1/2?=?693?h) and a negligible decay rate in the ability to transform substrate (D_r?=?0.08% h^?1). All these showed that this lipase had been effectively activated and stabilized by cotton fibres for the numerous hydroxyl groups and fluffy structure.     

Production of Linolenic Acid by Mortierella isabellina Grown on Octadecanol

The production of linolenic acid in mycelial lipids reached 0.31 mg/ml of culture broth when Mortierella isabellina was cultivated in a medium consisting of 2% octadecanol, 1% yeast extract, and 25 mmol/L of Mg²⁺ at 23°C for 5 days. Cultivation conditions were studied, and the results showed that (i) a suitable concentration of Mg²⁺ in the medium caused an increase in mycelial mass as well as linolenic acid production; (ii) when incubated at 23°C, maximal linolenic acid productivity was reached, although a higher content of the acid in total fatty acids was found at the lower temperature; (iii) the effect of substrate concentration on linolenic acid yield showed that the latter increased with concentration of substrate, and maximal linolenic acid yield was obtained with concentrations of 2% octadecanol and 1% yeast extract. Received: 27 November 2000 / Accepted: 22 June 2001     

Dehalogenation Activity of Selected Fungi Toward δ‐Iodo‐γ‐Lactone Derived from trans,trans‐Farnesol

Time‐course of biotransformation of racemic trans‐4‐((E)‐4′,8′‐dimethylnona‐3′,7′‐dien‐1‐yl)‐5‐iodomethyl‐4‐methyldihydrofuran‐2‐one ( 1 ) in fungal and yeast cultures was investigated. In these conditions, the substrate 1 was enantioselectively dehalogenated yielding 4‐((E)‐4′,8′‐dimethylnona‐3′,7′‐dien‐1‐yl)‐4‐methyl‐5‐methylenedihydrofuran‐2‐one ( 2 ) and its structure was established based on the spectroscopic data. The most effective biocatalyst used was Didymosphaeria igniaria, which catalyzed the process with highest rate and enantioselectivity (ee of product = 76%). The antiproliferative activity of δ‐iodo‐γ‐lactone 1 , product of its biotransformation 2 , and starting substrate (farnesol) were evaluated toward two cancer cell lines: A549 (human lung adenocarcinoma) and HL‐60 (human promyelocytic leukemia).     

Determination of substrate preferences of tench, Tinca tinca (L.), under controlled experimental conditions

The substrate preference of tench Tinca tinca (L.), 33 2‐year‐old individuals, mean size 10 ± 0.36 cm total length, was determined using four different substrates (concrete, artificial vegetation, sand and mud) in a 26 m long maquette. The species showed a high preference (100%) for the mud. Six more trials were performed combining the substrates by pairs (concrete–vegetation, concrete–mud, concrete–sand, mud–vegetation, mud–sand, sand–vegetation). Selection order was: mud (100% of the individuals when compared with concrete and sand and 98.26 ± 2.68% when compared with artificial vegetation), artificial vegetation (100% of the individuals when compared with concrete and 99.81 ± 0.76% when compared with sand), sand (98.29 ± 3.66% when compared with concrete) and concrete.     

Production,separation and purification of yeast invertase as a by-product of continuous ethanol fermentation

Summary A strain of Saccharomyces uvarum produced extracellular invertase in a chemostat reactor using a medium containing corn steep liquor and sugars. The production of yeast invertase increased with increase in corn steep liquor concentration. The production rate of invertase was maximal at a dilution rate of 0.75 h^–1. The production rate of invertase was found to be affected by the type of sugar substrate and fermentation temperature. The invertase in the crude broth could be purified by one-step DEAE chromatography. An 84% enzyme recovery with ninefold purification and 30-fold concentration could be achieved using this simple isolation procedure. Offsprint requests to: L. F. Chen     

Isolation of haemolytic bacilli from field‐collected Culicoides oxystoma and Culicoides peregrinus: potential vectors of bluetongue virus in West Bengal,India

Two haemolytic bacterial strains of Bacillus pumilus (CU1A, CU1B) and one blood‐utilizing strain of Bacillus licheniformis (CU2B) were isolated from relatively low numbers of field‐collected females of Culicoides oxystoma and Culicoides peregrinus (Diptera: Ceratopogonidae). A total of 36 females, including 18 of each of C. oxystoma and C. peregrinus (consisting of one and a pool of eight blood‐engorged specimens, and one and a pool of eight non‐engorged specimens for each species), were tested. In C. oxystoma, all three strains of bacteria were isolated from the one non‐engorged, the pool of non‐engorged and the pool of blood‐engorged females tested, but CU1A and CU2B were not found in the one blood‐engorged female tested. In C. peregrinus, all three strains were present in the pool of blood‐engorged females. However, the strain CU2B was not found in the pool of non‐engorged females. In the one blood‐engorged and one non‐engorged female tested, CU1A and CU2B were detected. The bacterial strains were identified based on Gram staining, enzyme activity (amylase and protease) and alignment of the 16S rRNA partial gene sequence to that available in the National Center for Biotechnology Information (NCBI) database GenBank. The functional role and significance of these haemolytic and blood‐digesting bacteria within the genus Culicoides remain to be determined.     

Feeding of fall armyworm,Spodoptera frugiperda,on Bt transgenic cotton and its isoline

Studies on insect food intake and utilization are important for determining the degree of insect/plant association and host species’ resistance, and also for helping design pest management programs by providing estimates of potential economic losses, techniques for mass breeding of insects, and identifying physiological differences between species. We studied the feeding and development of fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), on transgenic (Bt) and non‐transgenic (non‐Bt) cotton. The larvae of S. frugiperda fed on Bt cotton had a longer development period (23.0 days) than those fed on non‐Bt cotton (20.2 days). Survivorship of S. frugiperda larvae fed on Bt cotton (74.1%) was lower than that of larvae fed on non‐Bt cotton (96.7%). Pupal weight of larvae fed on Bt cotton (0.042 g) was lower than that of larvae fed on non‐Bt cotton (0.061 g). The cotton cultivar significantly affected food intake, feces production, metabolization, and food assimilation by S. frugiperda larvae. However, it did not affect their weight gain. Intake of Bt‐cotton leaf (0.53 g dry weight) per S. frugiperda larva was lower than the intake of non‐Bt‐cotton leaf (0.61 g dry weight). Larvae fed on Bt‐cotton leaves produced less feces (0.25 g dry weight) than those fed on non‐Bt‐cotton leaves (0.37 g dry weight). Weight gain per S. frugiperda larva fed on Bt‐cotton leaves (0.058 g dry weight) was similar to the weight gain for larvae fed on non‐Bt‐cotton leaves (0.056 g dry weight). The cotton cultivar significantly affected the relative growth, consumption, and metabolic rates, as well as other nutritional indices: the figures were lower for larvae fed on Bt‐cotton leaves than for larvae fed on non‐transgenic cotton leaves.     

Stilbella aciculosa: A Potential Biocontrol Fungus against Rhizoctonia solani

A semi‐solid fermentation product of the potential biocontrol fungus Stilbella aciculosa was formulated on wheat bran: water (1:1, w/w) and incubated 5, 10 and 15 days before addition to soil infested with the pathogen Rhizoctonia solani. Generally, preparations did not reduce survival of the pathogen in infested beet seed but they did prevent saprophytic growth of the pathogen from beet seed into soil. The magnitude of reduction by the 15‐day‐old inoculum was greater than that by the 5‐day‐old inoculum. Ten‐day‐old bran preparations of S. aciculosa at rates of 0.5 and 1.0% (w/w) in soil prevented post‐emergence damping‐off of cotton, radish and sugar beet in the glasshouse and a rate of 1.0% gave stands similar to those in the non‐infested control soil. The antagonist, grown on perlite formulated with molasses, cornmeal, alfalfa tissue or corn stover, prevented damping‐off of cotton in a naturally infested soil. However, the stands were not as great as that in soil planted with pentachloronitrobenzene (PCNB)‐treated seed. Toxic metabolites, produced by S. aciculosa developing on various substrates, slightly inhibited the growth of R. solani in culture and induced cytoplasmic leakage of the pathogen mycelium.     

Biocontrol of Fusarium and Verticillium Wilt of Tomato by Penicillium oxalicum under Greenhouse and Field Conditions

Treatments with conidia of Penicillium oxalicum produced in a solid‐state fermentation system were applied at similar densities (6 × 10⁶ spores/g seedbed substrate) to tomato seedbeds in water suspensions (T1: 5 days before sowing, or T2: 7 days before transplanting; 15 days after sowing), or in mixture with the production substrate (T3: 7 days before transplanting; 15 days after sowing). Treatments T2 and T3 significantly (P = 0.05) reduced fusarium wilt of tomato in both greenhouse (artificial inoculation) (33 and 28%, respectively) and field conditions (naturally infested soils) (51 and 72%, respectively), while treatment T1 was efficient only in greenhouse (52%). Verticillium wilt disease reduction was obtained with T3 in two field experiments (56 and 46%, respectively), while T1 and T2 reduced disease only in one field experiment (52% for both T1 and T2). Treatment with conidia of P. oxalicum plus fermentation substrate (T3) resulted in better establishment of a stable and effective population of P. oxalicum in seedbed soil and rhizosphere providing populations of approx. 10⁷ CFU/g soil before transplanting. Results indicate that it will be necessary to apply P. oxalicum at a rate of approx. 10⁶–10⁷ CFU/g in seedbed substrate and rhizosphere before transplanting for effective control of fusarium and verticillium wilt of tomato, and that formulation of P. oxalicum has a substantial influence on its efficacy.     

Response of multiple generations of beet armyworm,Spodoptera exigua (Hübner), feeding on transgenic Bt cotton

Development, reproduction and food utilization of three successive generations of beet armyworm, Spodoptera exigua (Hübner), fed on transgenic and non‐transgenic Bt cotton were examined. Significantly longer larval life‐span and lower pupal weight were observed in three successive generations of S. exigua fed on transgenic Bt cotton compared with non‐transgenic Bt cotton. Significantly higher survival rate and adult fecundity of S. exigua were found in three successive generations of S. exigua fed on transgenic Bt cotton compared with non‐transgenic Bt cotton. The survival rate and adult fecundity of S. exigua were occurred significant increase in the third generation compared with the first generation after feeding on transgenic Bt cotton. Significantly lower consumption, frass and relative growth rate (RGR) were observed in three successive generations of S. exigua fed on transgenic Bt cotton compared with non‐transgenic Bt cotton. Cotton variety significantly affected all indices of larval consumption and utilization in three successive generations of S. exigua, except for efficiency of conversion of ingested food. However, beet armyworm generation only significantly affected RGR of S. exigua. The results of this study indicated food quality on the diet‐utilization efficiency of S. exigua was different along with beet armyworm generation. Measuring multigenerational development and food utilization of S. exigua at individual and population level in response to Bacillus thuringiensis (Bt) can provide a more meaningful evaluation of long‐term population dynamics than experiments on a single generation. It is imperative to develop an appropriate multigenerational pest management tactic to monitor the field population dynamics of non‐target pests (e.g., beet armyworm) in agricultural Bt cotton ecosystem.     

Sublethal effects of Bt toxin and chlorpyrifos on various Spodoptera exigua populations

Bt cotton (Cry1Ac) has been commercially grown in China since 1997, saving China's cotton production from attack by Bt‐target pests and also tremendously reducing pesticide usage. In recent years, however, Bt cotton, with 4.2 million ha of cultivation, has suffered from a secondary target pest, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). In China, growers have even had to re‐adopt conventional pesticides to control the pest, and this practice has already caused serious pesticide residue. In order to clarify the sublethal effects of chemical pesticide, the responses of a Bt‐susceptible and a Bt‐tolerant (Bt10) S. exigua strain to three treatment combinations were examined, including Bt toxin, sublethal chlorpyrifos, and Bt + sublethal chlorpyrifos. The susceptible and the Bt10 strain responded differently to dual pressure. Bt toxin + sublethal chlorpyrifos treatment lowered larval mortality and stimulated population increase of the susceptible S. exigua, whereas it delayed growth and development of the Bt10 strain. Under dual pressure, although larvae of the Bt10 strain developed faster than larvae of the susceptible strain, the Bt10 population experienced higher larval mortality, prolonged pupal duration, decreased pupal weight, decreased emergence rate, and shortened adult longevity. Compared with the susceptible strain, the Bt10 strain was deleteriously affected by sublethal chlorpyrifos. The Bt‐tolerant/resistant S. exigua population was more vulnerable to chemical pesticides like chlorpyrifos regardless of whether it was exposed to Bt toxin or not. Our study provides a reference for increasing the efficacy of control of S. exigua in Bt‐cotton planting areas.     

Aerosol distribution and efficacy in a commercial food warehouse

A series of field trials were conducted in a commercial food storage facility to evaluate exposure of stored‐product insects to aerosol formulations of synergized pyrethrins and the insect growth regulator methoprene. When adults of Tribolium castaneum (Herbst), the red flour beetle, and Tribolium confusum (Jacqueline DuVal), the confused flour beetle were exposed with and without a food source to synergized pyrethrin aerosol, there was no difference in adult mortality with respect to availability of food at either 7 or 14 days after exposure (P≥ 0.05). However, mortality was lower in T. confusum (40.4% and 79.3% with flour at 7 and 14 days, 38.9% and 84.8% without flour at 7 and 14 days) compared to T. castaneum (96.5% and 99.8% with flour at 7 and 14 days, 91.0% and 98.7% without flour at 7 and 14 days). Few late‐stage larvae and pupae of either species exposed to the pyrethrin aerosol emerged as adults. In tests with methoprene aerosol, adult emergence of exposed 3‐and 4‐week‐old larvae of T. confusum was less than 2%. Only 0.3% of 4‐week‐old larvae of T. castaneum exposed in open and obstructed areas emerged as adults. Emergence of adults from eggs of Plodia interpunctella (Hübner), the Indianmeal moth, embedded in culture media and exposed to the methoprene aerosol was 13.2%± 3.5%. Results show that the aerosols evaluated in our study could give effective control of some of the major stored‐product insect pests in commercial food storage facilities, and may offer an alternative to fumigation.     

Effects of Three‐Hour Restricted Food Access during the Light Period on Circadian Rhythms of Temperature,Locomotor Activity,and Heart Rate in Rats

The effects of food on biological rhythms may influence the findings of chronopharmacological studies. The present study evaluated the influence of a restricted food access during the rest (light) span of nocturnally active Wistar rats on the 24 h time organization of biological functions in terms of the circadian rhythms of temperature (T), heart rate (HR), and locomotor activity (LA) in preparation for subsequent studies aimed at evaluating the influence of timed food access on the pharmacokinetics and pharmacodynamics of medications. Ten‐wk‐old male Wistar rats were housed under controlled 12:12 h light:dark (LD) environmental conditions. Food and water were available ad libitum, excepted during a 3 wk period of restriction. Radiotelemetry transmitters were implanted to record daily rhythms in T, HR, and LA. The study lasted 7 wk and began after a 21‐d recovery span following surgery. Control baseline data were collected during the first wk (W1). The second span of 3 wk duration (W2 to W4) consisted of the restricted feeding regimen (only 3 h access to food between 11:00 and 14:00 h daily) during the L (rest span) under 12:12 h LD conditions. The third period of 3 wk duration (W5 to W7) consisted of the recovery span with ad libitium normal feeding. Weight loss in the amount of 5% of baseline was observed during W1 with stabilization of body weight thereafter during the remaining 2 wk of food restriction. The 3 h restricted food access during the L rest span induced a partial loss of circadian rhythmicity and the emergence of 12 h rhythms in T, HR, and LA. Return to ad libitum feeding conditions restored circadian rhythmicity in the manner evidenced during the baseline control span. Moreover, the MESORS and amplitudes of the T, HR, and LA 24 h patterns were significantly attenuated during food restriction (p<0.001) and then returned to initial values during recovery. These changes may be interpreted as a masking effect, since T, HR, and LA are known to directly react to food intake. The consequences of such findings on the methods used to conduct chronokinetic studies, such as the fasting of animals the day before testing, are important since they may alter the temporal structure of the organism receiving the drug and thereby compromise findings.     

Production of aflatoxin by Aspergillus parasiticus NRRL-2999 during growth in the presence of curing salts

Aspergillus parasiticus NRRL-2999 was inoculated into meat mixtures with curing salts and into yeast extractsucrose (YES) and sucrose-ammonium salts (SAS) broth with and without curing salts to determine if the presence of curing salts significantly affected growth and aflatoxin production by the mold. The effect of individual curing salts or curing salt mixtures on growth and toxin elaboration by the aspergillus was substrate dependent. When YES broth contained 100 ppm of NaNO₂, 2% NaCl, or 1 or 2% NaCl plus 200 ppm of NaNO₂ or 200 ppm of NaNO₃, growth and/or aflatoxin production was depressed. Biosynthesis of aflatoxin B₁ was enhanced by presence of 1 and 4% NaCl in YES broth. The SAS broth containing only NaCl or NaCl combined with nitrite or nitrate yielded less aflatoxin than did control broth or no aflatoxin at all. When compared to the control, an increase in growth and amount of aflatoxin occurred in SAS broth which contained 200 ppm of NaNO₃. Sausages containing 100 and 200 ppm NaNO₂ and no NaCl supported more mold growth and aflatoxin production than did control sausage with 3 % NaCl and 100 ppm of NaNO₂. Addition of 2 and 3 % NaCl and no nitrite to sausage resulted in less aflatoxin than in control sausage.