Inhibition in aflatoxin biosynthesis by the extract of Amorphophallus campanulatus (OL) and calcium oxalate

A complete inhibition in aflatoxin production by Aspergillus flavus was observed with greater than 4.5 mg ml^-1 of 20 g 100 ml^-1 leaf extract of Amorphophallus campanulatus (OL). Suppression in growth was also well pronounced (81% at 8.0 mg ml^-1). The comparative efficacy of the corm of the plant was lower. Calcium oxalate, an important constituent of 'OL', completely checked the growth and toxin biosynthesis at 0.4 mg ml^-1 concentration.     

Effect of spice essential oils on growth and aflatoxin B1 production by Aspergillus flavus

Aflatoxin B¹ production by Aspergillus flavus was studied in yeast extract sucrose broth in the presence of cinnamon, clove, almond and cardamom oils. Growth and aflatoxin B₁ production was inhibited by 0.5 μl cinnamon oil ml^-1 medium and by 1 μl clove oil ml_-1. Almond and cardamom oils only affected growth when their concentration exceeded 1.25 μl ml^-1 medium. Aflatoxin B₁ production was stimulated by 0.75 and 1 μl almond oil ml^-1 medium or by 0.25 and 0.5 μl cardamom oil ml^-1.     

Effect of Iysozyme concentration, heating at 90°C, and then incubation at chilled temperatures on growth from spores of non-proteolytic Clostridium botulinum

The heat treatment necessary to inactivate spores of non-proteolytic Clostridium botulinum in refrigerated, processed foods may be influenced by the occurrence of lysozyme in these foods. Spores of six strains of non-proteolytic Cl. botulinum were inoculated into tubes of an anaerobic meat medium, to give 10⁶ spores per tube. Hen egg white lysozyme (0–50 μg ml^-1) was added, and the tubes were given a heat treatment equivalent to 19·8 min at 90°C, cooled, and incubated at 8°, 12°, 16° and 25°C for up to 93 d. In the absence of added lysozyme, neither growth nor toxin formation were observed. A 6–D inactivation was therefore achieved. In tubes to which lysozyme (5–50 μg ml^-1) had been added prior to heating, growth and toxin formation were observed. With lysozyme added at 50 μg ml^-1, growth was first observed after 68 d at 8°C, 31 d at 12°C, 24 d at 16°C, and 9 d at 25°C. Thus, in these circumstances, a heat treatment equivalent to 19·8 min at 90°C was not sufficient, on its own, to give a 6–D inactivation. A combination of the heat treatment, maintenance at less than 12°C, and a shelf-life not more than 4 weeks reduced the risk of growth of non-proteolytic Cl. botulinum by a factor of 10⁶.     

Comparison of isopropyl cinodine with nalidixic acid in the direct viable count

Isopropyl cinodine and nalidixic acid were compared in the direct viable count. With raw water and biofilms, elongated cells were seen in the presence of isopropyl cinodine. Increased incubation time led to an increased direct viable count. Individual bacteria responded differently to isopropyl cinodine. Five organisms grew in the presence of 0.01 μg ml^-1 of isopropyl cinodine but were inhibited by 0.1 μg ml^-1. These values for a sixth organism were 0.1 μg ml^-1 and 1.0 μg ml^-1 respectively. The direct viable count was done with inocula taken when the cells were in either lag, log or stationary phases of growth. No differences were seen in the percentage of elongated cells within an experiment but there was variation between experiments. The effect of nalidixic acid and isopropyl cinodine appeared to be additive with respect to inhibition of growth, but little or no additive effect was seen upon the percent of nutrient responsive cells.     

The enzymatic reaction for the production of panose and isomaltose by glucosyltransferase from Aureobasidium

Glucosyltransferase from Aureobasidium produced 212 mg ml^-1 of glucosyl-oligosaccharides (panose: Glcα1→6Glcα1→4Glc 189 mg ml^-1 and isomaltose: Glcα1→6Glc 23 mg ml^-1) from maltose: Glcα1→4Glc at a high concentration (500 mg ml^-1) and the efficiency of production was 42-4%. The enzymatic reaction from maltose to panose is reversible but that from panose to isomaltose is not.     

Anaerobic degradation of pectin by mixed consortia and optimization of fermentation parameters for higher pectinase activity

Samples from biogas digesters, sewage ponds, animal house effluents and food processing wastes were used in enrichment systems seeking anaerobic bacteria producing pectinases. Among the 46 anaerobic consortia developed from various samples, four showed high pectinase activity under static anaerobic conditions. Investigation of fermentation variables showed the optimum conditions for pectinase activity were pH 7.0, 45°C and 72 h of growth with 0.5% pectin in the cultivation medium. A 1.4- to 1.6-fold increase in the pectinase activity was achieved under these conditions. The maximum yield of enzymes (62.72 U ml-1 of pectinase, 4.74 U ml^-1 of polygalacturonase, 113.30 U ml^-1 of pectin lyase, 2.10 U ml^-1 of pectinesterase, 0.75 U ml^-1 of total cellulase and 9.27 U ml^-1 of xylanase) was recorded with the consortia C-S2 developed from decomposed plant samples collected from a pond.     

Effect of some factors on determination of viable microbial cells by peroxyoxalate chemiluminescence method

The effects of physical and chemical factors on the production of H₂O₂ from Escherichia coli cells were studied. When 20 mmol 1^-1 Tris-HCl buffer was used for this purpose the electron transport system (ETS) showed the highest activity at pH 7.6-8.2. KCN promoted the production of H₂O₂ from E. coli cells, and the optimum concentration was changed in different reaction times and pH values. Glucose, 5 mg ml^-1, increased the ETS activity about twofold. The other substrates and surfactants did not increase the chemiluminescence intensity. NaNO₂ and Na₂SO₄ in inorganic salts significantly reduced the ETS activity above 70%. In addition, the optimum temperature for the production of H₂O₂ was 30°C in this study. When glucose (5 mg ml^-1) and KCN (0.2 mmol 1^-1) were added to the reaction buffer containing 0.5 mmol 1^-1 menadione, the detectable minimum cell densities (averages of triplicate assay) of E. coli, Enterobacter cloacae and Serratia marcescens were 5 times 10³ cells ml^-1, 10⁴ cells ml^-1 and 10⁴ cells ml^-1 respectively.     

The influence of pH and growth rate on production of the bacteriocin, bavaricin MN, in batch and continuous fermentations

Bavaricin MN, a bacteriocin produced by Lactobacillus bavaricus MN, reached titres of 2000 AU ml^-1 in APT broth maintained at pH 6.0, 30°C in a batch fermenter. Levels of bavaricin MN at pH 5.5 and 6.5 were lower despite comparable levels of producer cells. The addition of 3.0 g l^-1 beef extract to APT broth resulted in increases in both the growth rate of the culture and the production of bavaracin MN. The titre of bavaricin MN in batch fermenters controlled at pH 6.0 in APT broth plus 3 g l^-1 beef extract reached 3200 AU ml^-1 at 30°C. This level was reduced to 800 AU ml^-1 by 76 h. Glucose-limited continuous culture of Lact. bavaricus MN under the same conditions resulted in an increase in the titre of bavaricin MN to 6400 AU ml^-1. This level was maintained, independent of growth rate, for 345 h. Growth rates of 0.205, 0.118, 0.169 and 0.058 h^-1 were examined.     

Efflux of ions and ATP depletion induced by pediocin PA-1 are concomitant with cell death in Listeria monocytogenes Scott A

Domination of Carnobacterium divergens LV13 by a bacteriocin-producing (bac⁺) organism Carnobacterium piscicola LV17 was dependent on the level of inoculum of the producer strain and its bacteriocin production. When C. piscicola LV17 was grown in APT broth from an initial inoculum of α-10⁴ cfu ml^-1, bacteriocin was not produced (bac^-) although maximum population was reached. The culture remained bac^- during subsequent inoculation at 10²-10⁷ cfu ml^-1 unless it was first grown on solid medium or if heat-treated supernatant fluids from a bac⁺ culture of C. piscicola LV17, LV17A or LV17B were added to the culture prior to the stationary phase of growth. Use of purified carnobacteriocins from C. piscicola LV17A and LV17B confirmed their role in regulation of the bac⁺ phenotype. The need for induction might account in part for differences in bacteriocin production by cultures in liquid and on solid growth media.     

Antibacterial activity of anacardic acid and totarol, alone and in combination with methicillin, against methicillinresistant Staphylococcus aureus

The inhibitory and bactericidal activities of anacardic acid and totarol, alone and in combination with methicillin, were investigated against methicillin-resistant Staphylococcus aureus (MRSA). The growth of two MRSA strains was inhibited by 6·25 μg ml^-1 of anacardic acid and 0·78 μg ml^-1 of totarol. The time-kill curve study showed that these two compounds were bactericidal against MRSA. Anacardic acid killed MRSA cells more rapidly than totarol, and no viable cells were detected after being exposed to 6·25 μg ml^-1 of anacardic acid for 6 h. Anacardic acid showed bactericidal activity against MRSA at any stage of growth, and also even when cell division was inhibited by chloramphenicol. In the combination studies, the minimal inhibitory concentration (MIC) of methicillin was lowered from 800 to 1·56 μg ml^-1 for MRSA ATCC 33591, and from 800 to 6·25 μg ml^-1 for MRSA ATCC 33592, by combining with 1/2 X MIC of anacardic acid. The time-kill curves demonstrated synergistic bactericidal activities for these combinations.     

New selective media for isolation of clostridia from faecal specimens

Two selective media, novobiocin-colistin agar (NCA) and colistin-crystal violet agar (CCA), were developed for isolating clostridia from human and animal faeces. The basal medium was modified Eggerth-Gagnon agar. The NCA medium contains novobiocin (8 μg ml^-1) and colistin (8 μg ml^-1) and the CCA medium contains colistin (10 μg ml^-1) and crystal violet (10 μg ml^-1). Nine faecal specimens were cultured. Clostridia isolated on these media were similar to those on non-selective media, and higher than those isolated after heat treatment. However, more clostridial species were isolated on the new selective media compared with the non-selective medium. These selective agars were particularly useful for enumerating and isolating clostridia from human faeces.     

The detection of Salmonella in skimmed milk powder enrichments using conventional methods and immunomagnetic separation

Skimmed milk powders were spiked with one of three Salmonella serovars and incubated in buffered peptone water for 24 h. No false-negative results were obtained by immunomagnetic separation (IMS), compared to seven for selenite cysteine, one for Müller-Kauffmann tetrathionate and two for Rappaport-Vassiliadis enrichment broths. Salmonella virchow was detected and enumerated during the pre-enrichment incubation by IMS and indirect conductance techniques. The Salm. virchow cell number did not increase after 12 h incubation and remained at 3 times 10⁶ cfu ml^-1. IMS was able to capture Salm. virchow cells at cell numbers ca 50 ml^-1 in the presence of a 1000 greater number of non-salmonella cells.     

The effect of clove and cinnamon oils on growth of and aflatoxin production by Aspergillus flavus

The effect of different concentrations of clove and cinnamon oils was studied on the growth of and aflatoxin production by Aspergillus flavus in SMKY liquid medium. The effect of these compounds was also verified against aflatoxin production in maize. Significant reduction (P < 0.05) in the elaboration of aflatoxin in liquid culture after treatment with more than 100 μg ml^-1 of these compounds was recorded. Cinnamon oil exhibited maximum inhibitory action and reduced 78% aflatoxin formation on maize at 1000 mg kg^-1.     

Sensitivity of Bacillus coagulans spores to combinations of high hydrostatic pressure, heat, acidity and nisin

We investigated the combined effects of pressure, temperature, pH, initial spore concentration and the presence of nisin on the survival of spores of Bacillus coagulans. Spores were more sensitive to pressure both at lower pH and at higher treatment temperatures. An additional 1.5-log₁₀ reduction in cfu ml^-1 was observed when pH was lowered from 7.0 to 4.0 during pressurization at 400 Mpa and 45°C. A 4-log₁₀ cfu ml^-1 reduction was observed when the temperature was increased from 25°C to 70°C during pressurization at 400 Mpa. The spores were sensitive to nisin at concentrations as low as 0.2 IU ml^-1. At least a 6-log₁₀ reduction was generally achieved with pressurization at 400 Mpa in pH 4.0 buffer at 70°C for 30 min when plated in nutrient agar containing 0.8 IU ml^-1 nisin.     

Polyamine biosynthesis in the ectomycorrhizal fungus Paxillus involutus exposed to zinc

Following exposure of the ectomycorrhizal fungus Paxillus involutus to 3·75 μg ml^-1 or 15 μg ml^-1 zinc for 2 weeks, growth was unaffected, putrescine and spermidine concentrations remained unaltered, while spermine concentration increased substantially. This was accompanied by small changes in the activities of ornithine decarboxylase and S -adenosylmethionine decarboxylase. There was, however, reduced synthesis of aminopropylcadaverine and N,N' bis (3-aminopropyl)cadaverine at the higher zinc concentration. It is possible that the increased spermine concentration may have resulted from displacement of the amine from the cell wall by the metal ion.     

Growth and growth substrate levels in spinach under non-steady state conditions of nitrogen nutrition and light

Spinach plants ( Spinacia oleracea L. cv. Subito) were grown in a complete nutrient solution under ample light intensity (14 h day⁻¹ at 660 μmol m⁻² s⁻¹) before being transferred either to a minus-N solution (experiment 1), or to limiting light conditions (6 h day⁻¹ at 220 μmol m⁻² s⁻¹; experiment 2). Shoot growth in experiment 1 decreased significantly from 0.24 day⁻¹ to 0.07 day⁻¹ after the fourth day of transfer. Root relative growth rate increased after 1 day from 0.25 to 0.31 day⁻¹, but decreased on the fifth day after transfer to 0.11 day⁻¹. Shoot growth in experiment 2 decreased significantly from 0.25 to 0.17 day⁻¹ after the fourth day of transfer, while root growth decreased to half of its original level (0.25 day⁻¹) already on the second day. Growth substrate levels in the plants (free sugars, free amino acids) and starch levels depended on the plant age, the moment in the diurnal cycle, and the imposed treatment. Fluctuations in shoot growth or root growth resulting from the light or N limitation could not be explained by a correspondent increase or decrease in the levels of growth substrates. The hypotheses underlying the functional equilibrium theory, assuming shoot and root growth to be controlled by N- and C-containing substrates respectively, and several other growth and partitioning models are therefore questioned. A neglect of the osmotic role of the free sugars in these models might be the explanation for this.     

Growth and Grazing Rates of the Marine Planktonic Ciliate Strombidinopsis sp. on Red-Tide and Toxic Dinoflagellates

ABSTRACT We investigated growth and grazing rates of Strombidinopsis sp. when feeding on several species of red-tide and/or toxic dinoflagellates. Strombidinopsis sp. one of the largest aloricate choreotrichs so far reported, grew well on Lingulodinium polyedrum, Gymnodinium sanguineum, Scrippsiella trochoidea, Cochlodinium polykrikoides , and Prorocentrum minimum , but failed to grow on Amphidinium carterae. Specific growth rates of Strombidinopsis sp. increased rapidly with increasing prey density up to ca. 100 ng C ml^-1, but were saturated or increased slightly at higher concentrations. Maximum specific growth rates of Strombidinopsis sp. on various prey species were 1.38 day^-1 for C. polykrikoides , 1.27 for G. sanguineum , 1.06 for P. minimum , 0.83 for L. polyedrum , and 0.67 for S. trochoidea. Threshold prey concentrations (where net growth = 0) were 12–38 ng C ml^-1. Maximum ingestion and clearance rates of Strombidinopsis sp. were 353 ng C grazer^-1 day^-1 and 110 μ, l grazer^-1 h^-1, respectively. Strombidinopsis sp. exhibited higher maximum growth, ingestion, and clearance rates than the mixotrophic dinoflagellate Fragilidium cf. mexicanum or the heterotrophic dinoflagellates Protoperidinium cf. divergens and P. crassipes , when grown on the same prey species. In addition, the sequence of prey species arranged according to growth response of Strombidinopsis sp. differed considerably from those of Fragilidium cf. mexicanum, Protoperidinium cf. divergens , and P. crassipes.     

Purification and characterization of glucoamylase produced by Aspergillus niger in solid state fermentation

Glucoamylases produced by Aspergillus niger grown on wheat bran in solid cultures were purified. Four different forms, GA I, GA I', GA II and GA III, were found having apparent molecular weights of 112 000, 104 000, 74 000 and 61 000 Da respectively. The enzymes are glycoproteins with a carbohydrate content of 16%, and optimal activity at 60C and pH 4.4. Activity was strongly inhibited by Hg²⁺ while Mn²⁺ and Fe²⁺ were stimulatory. The K_m values for the degradation of starch and maltose were 3.5 and 7.8 mg ml^-1, respectively.     

Comparison of conductimetric and traditional plating techniques for detecting yeasts in fruit juices

Frozen fruit juice concentrates containing an average microbial population of log₁₀ 1.54 cfu ml^-1 were examined by traditional plating techniques and direct and indirect conductimetry. The initial populations in diluted (1:4) concentrates increased to an average of log₁₀ 3.82 cfu ml^-1 during incubation at 25°C for 24 h. Incubation before plating and subjecting to conductimetric tests also facilitated the resuscitation of cells that may have been freeze-injured. Yeasts were recovered in equal numbers on acidified (pH 3.5) potato dextrose agar and dichloran rose bengal chloramphenicol agar (pH 5.6). Yeasts and bacteria were recovered on orange serum agar. Detection times determined by indirect conductimetry correlated fairly well ( r = -0.73) with populations (cfu ml^-1) detected on traditional plating media. Populations in diluted concentrates which were not incubated before examination were detected conductimetrically in an average of 48.9 h, whereas detection times for diluted concentrates incubated for 24 h at 25°C before testing were reduced to an average of 14.1 h. Examination by conventional (direct) conductimetry required an additional 10–20 h to reach changes in conductance of 5 μS h^-1.     

Interaction between the effects of bacteria and dry storage on the opening and water relations of cut rose flowers

W.G. VAN DOORN AND K. D'HONT. 1994. Flowering stems of four rose cultivars (Sonia, Madelon, Jacaranda and Frisco) were placed in aqueous suspensions of bacteria at 10⁴ and 10⁸ colony-forming units (cfu) ml^-1 for 24 h at 5C, then stored dry or held in water for 24 h at 8C and subsequently placed in vase-water at 20C. The effects of these treatments on vase-water uptake were similar to the effects on flower opening. In Sonia and Madelon roses flower opening was negatively affected both by 10⁸ cfu ml^-1 of bacteria and by dry storage. No effect was found at 10⁴ cfu ml^-1, but this concentration had a detrimental effect on flower opening when combined with dry storage. Although flower development in Jacaranda roses was not affected by the bacteria treatments it was inhibited by dry storage. This inhibition was progressively greater when the stems had previously been pulse-treated with a larger number of bacteria. Flower opening in Frisco roses was not affected by even the highest concentration of bacteria, nor by the period of dry storage. It is concluded that placing flowers in water containing bacteria (up to 10⁸ cfu ml^-1) may not always have a negative effect on flower development in cut rose flowers but, together with the effects of dry storage, the presence of even a low number of exogenous bacteria (10⁴ cfu ml^-1) inhibits the development in several cultivars. Such bacterial counts are nearly always found in samples of water used for standing roses during distribution to the consumers.