Biomedical subjects

V Sanchis

Publications and source records attributed to V Sanchis.

At least 55 records · Page 3Linked to original sources

PubMed1999

Control of growth and fumonisin B1 production by Fusarium verticillioides and Fusarium proliferatum isolates in moist maize with propionate preservatives.

The effect of propionic acid, its sodium salt or a commercial formulation of propionates (0.03, 0.05 and 0.07%), on growth and fumonisin B1 production by Fusarium verticillioides and F. proliferatum isolated was evaluated on irradiated maize at different water activities (aw, 0.93, 0.95, 0.98) and temperatures (15, 25 degrees C). The four isolates grew at all aw x temperature treatments in the absence of propionates. At the highest propionate concentration tested (0.07%), however, growth was restricted to 0.98 aw, for F. proliferatum isolates but not for those of F. verticillioides. Inhibition of growth was maximum when propionates were added in the acid form. In the presence of low propionate concentrations (0.03%), growth was sometimes enhanced probably due to assimilation of these compounds by the fungus. Water activity, temperature, concentration and source of propionate, as well as most two-, three-, four-, and five-way interactions had a significant influence on growth of Fusarium isolates. None of the assayed treatments had any effect on fumonisin B1 production by F. verticillioides isolates. For F. proliferatum, higher fumonisin B1 production occurred in the absence of propionates, and in general concentration decreased with increasing doses of preservatives. Single factors (aw, propionate concentrations and temperature) and temperature x aw and propionate concentration x temperature interactions had a significant effect on fumonisin production (p < 0.01). Moreover, propionate concentration was the single most important factor, besides temperature, which affected fumonisin B1 production.

Carboxylic Acids↗

PubMed1999

Development and field performance of a broad-spectrum nonviable asporogenic recombinant strain of Bacillus thuringiensis with greater potency and UV resistance.

The main problems with Bacillus thuringiensis products for pest control are their often narrow activity spectrum, high sensitivity to UV degradation, and low cost effectiveness (high potency required). We constructed a sporulation-deficient SigK(-) B. thuringiensis strain that expressed a chimeric cry1C/Ab gene, the product of which had high activity against various lepidopteran pests, including Spodoptera littoralis (Egyptian cotton leaf worm) and Spodoptera exigua (lesser [beet] armyworm), which are not readily controlled by other Cry delta-endotoxins. The SigK(-) host strain carried the cry1Ac gene, the product of which is highly active against the larvae of the major pests Ostrinia nubilalis (European corn borer) and Heliothis virescens (tobacco budworm). This new strain had greater potency and a broader activity spectrum than the parent strain. The crystals produced by the asporogenic strain remained encapsulated within the cells, which protected them from UV degradation. The cry1C/Ab gene was introduced into the B. thuringiensis host via a site-specific recombination vector so that unwanted DNA was eliminated. Therefore, the final construct contained no sequences of non-B. thuringiensis origin. As the recombinant strain is a mutant blocked at late sporulation, it does not produce viable spores and therefore cannot compete with wild-type B. thuringiensis strains in the environment. It is thus a very safe biopesticide. In field trials, this new recombinant strain protected cabbage and broccoli against a pest complex under natural infestation conditions.

Animals↗

PubMed1998

Colonisation and competitiveness of Aspergillus and Penicillium species on maize grain in the presence of Fusarium moniliforme and Fusarium proliferatum.

The effects of different steady-state water activity levels (a(w), 0.93, 0.95 and 0.98) and temperature (15 and 25 degrees C) on colonisation patterns of Aspergillus and Penicillium spp., when colonising irradiated maize grain in the presence of Fusarium moniliforme and Fusarium proliferatum were assayed in terms of populations (colony forming units, CFUs g grain(-1)), seed infection and colonisation rates. The activity of F. moniliforme and F. proliferatum in grain reduced the presence of Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus to some extent, particularly at 15 degrees C and higher water availabilities (0.95-0.98 a(w)). In contrast, colonisation patterns of Penicillium implicatum on maize grain were unaffected by either Fusarium spp. in terms of CFUs or seed infection. Correlations were made between CFUs, seed infection, growth rates and niche overlap indices and hyphal interactions to try and link key indicators of competitiveness and dominance by an individual species.

Aspergillus↗

PubMed1998

Effect of water activity and temperature on growth and ochratoxin production by three strains of Aspergillus ochraceus on a barley extract medium and on barley grains.

Aspergillus ochraceus Wilhelm is a widespread storage fungus that has been isolated from grains such as barley. The objective of this study was to determine the effects of water activity (a(w)), temperature, time and their interactions on (a) growth on a barley extract agar medium, (b) growth on barley grains and (c) ochratoxin production on barley grains by three strains of A. ochracieus. For the three A. ochreceus isolates examined (NRRL 3174, 3.113 and 3.38), optimal a(w) levels for growth on agar media were in the range 0.98-0.96, with temperature optima of 30 degrees C for two of the isolates and 25-30 degrees C for the other isolate. Growth was observed at 10 and 37 degrees C, but only at higher a(w) levels assayed. Two dimensional profiles were constructed for the range of a(w) and temperature conditions that allow growth of the three isolates. Maximum growth on barley grains was reached at 30 degrees C, at both a(w) levels assayed (0.96 and 0.98), with fungal growth rates in the 4-5 mm day(-1) range. Maximum amounts of ochratoxins were produced at the highest a(w) treatment (0.98 a(w)) and after a three-week incubation time at 25-30 degrees C. The range of ochratoxin concentrations varied considerably, from 1.7 to 12,949 ppm, depending on the a(w) and temperature interactions.

Aspergillus ochraceus↗

PubMed1998

Effect of water activity on hydrolytic enzyme production by Fusarium moniliforme and Fusarium proliferatum during colonisation of maize.

The effect of different water availabilities (water activity, aw; 0.98-0.93) and time (up to 15 days) on the production of seven hydrolytic enzymes by strains of F. moniliforme and F. proliferatum during early colonisation of gamma-irradiated living maize grain were examined in this study. Both the total activity (micromol 4-nitrophenol min(-1) g(-1) maize) and specific activity (nmol 4-nitrophenol min(-1) microg(-1) protein) were quantified using chromogenic p-nitrophenyl substrates. The dominant three enzymes produced by the fungi on whole colonised maize kernels were alpha-D-galactosidase, beta-D-glucosidase, and N-acetyl-beta-D-glucosaminidase. The other four enzymes were all produced in much lower total amounts and in terms of specific activity (beta-D-fucosidase, alpha-D-mannosidase, beta-D-xylosidase and N-acetyl-alpha-D-glucosaminidase), similar to that in uncolonised control maize grain. There were significant increases in the total production of the three predominant enzymes between 3-15 days colonisation, and between 3-6 days in terms of specific activity when compared to untreated controls. The total and specific activity of the alpha-D-galactosidase, beta-D-glucosidase and N-acetyl-beta-D-glucosaminidase, were maximum at 0.98 aw with significantly less being produced at 0.95 and 0.93 aw, with the exception of the total activity of alpha-D-galactosidase which was similar at both 0.95 and 0.93 aw. Single factors (time, aw, and inoculation treatment), two- and three- way interactions were all statistically significant for the three dominant enzymes produced except for specific activity of beta-D-glucosidase (two and three-way interactions) and for total activity of alpha-D-galactosidase in the time x aw treatment. This study suggests that these hydrolytic enzymes may play an important role in enabling these important fumonisin-producing Fusarium spp. to rapidly infect living maize grain over a wide aw range.

Acetylglucosaminidase↗

PubMed1998

Biological control of major postharvest pathogens on apple with Candida sake.

Epiphytic microorganisms isolated from apples, pears and the surfaces of apple leaves were screened for antagonistic activity against Penicillium expansum (blue-mold), Botrytis cinerea (gray-mold) and Rhizopus nigricans (Rhizopus rot) on apple (Malus domestica). A total of 933 bacteria and yeasts were tested in primary screening against P. expansum. Ninety-two strains reduced the lesion size on apples by more than 50%, 72 of which were isolated from the surface of apples. For secondary screening against P. expansum, B. cinerea and R. nigricans, 31 strains were selected. The most promising isolate, CPA-1, was identified as Candida sake. This yeast, isolated from apples in storage season was very effective against all three diseases. Wounded Golden Delicious apples protected with the yeast suspension at a concentration of 2.6 x 10(6) CFU/ml and inoculated with conidia of B. cinerea and R. nigricans of 10(5) and 10(4) conidia/ml, respectively, did not develop rot. Complete control of P. expansum was obtained at the same concentration of the antagonist with a pathogen inoculum concentration of 10(3) conidia/ml. This strain, also provided excellent control of rot development under cold storage conditions. The strain of Candida sake can grow actively in aerobic conditions. In drop-inoculated wounds of apples, the populations of C. sake increased by more than 50-fold during the first 24 h at 20 degrees C. The maximum population of C. sake on apple wounds was the same at 20 as at 1 degrees C and was recovered after three and twenty days, respectively.

Candida↗

PubMed1998

Occurrence of fumonisins in Spanish beers analyzed by an enzyme-linked immunosorbent assay method.

This study describes the occurrence of fumonisin in a survey of 32 Spanish beers, including some non-alcoholic beers. The analysis was conducted using an enzyme-linked immunosorbent assay (ELISA) method and, in the range assayed (0-100 ng fumonisin B1/ml spiked-beer), recovery rates were about 81%, with variation coefficients that did not exceed 4.2%. The detection limit with this method was 3 ng fumonisins/ml beer. Results for the fourteen positive samples (43.8% of total samples) showed that the lowest amount of fumonisins found in a positive sample was 4.76 ng/ml and the highest was 85.53 ng/ml.

Beer↗

PubMed1998

Ecological determinants for germination and growth of some Aspergillus and Penicillium spp. from maize grain.

This study compared the effect of temperature (5-45 degrees C), water availability (water activity, aw; 0.995-0.75) and their interactions on the temporal rates of germination and mycelial growth of three mycotoxigenic strains of Aspergillus ochraceus and one isolate each of A. flavus, A. niger, Penicillium aurantiogriseum and P. hordei in vitro on a maize extract medium. Germination was very rapid at > 0.90 aw with an almost linear increase with time for all species. However, at < 0.90 aw, the germination rates of A. flavus and P. hordei were slower. The aw minima for germination were usually lower than for growth and varied with temperature. The effect of aw x temperature interactions on the lag phases (h), prior to germination, and on the germination rates (h(-1)), were predicted for the first time for these fungi using the Gompertz model modified by Zwietering. This showed that A. flavus, A. niger and the two Penicillium spp. had very short lag times between 0.995-0.95 aw over a wide temperature range. At marginal temperatures, these were significantly higher, especially at < 10 degrees C for Aspergillus spp. and > 30 degrees C for Penicillium spp. There were also statistically significant differences between lag phases and germination rates for three different isolates of A. ochraceus. The Aspergillus spp. also germinated faster than the Penicillium spp. The temperature x aw profiles for mycelial growth varied considerably between species, both in terms of rates (mm d(-1)) and tolerances. Predictions of the effects of important environmental factors such as temperature, aw and their interactions on lag times to germination, germination rates and mycelial growth are important in the development of hurdle technology approaches to predicting fungal spoilage in agricultural and food products.

Aspergillus↗

PubMed1998

Colonization of maize grain by Fusarium moniliforme and Fusarium proliferatum in the presence of competing fungi and their impact on fumonisin production.

This study was carried out to determine the effect of water activity (aw) and temperature on the patterns of colonization of maize grain by isolates of Fusarium moniliforme and F. proliferatum in the presence of interacting spoilage fungi, such as Aspergillus flavus, A. niger, A. ochraceus, and Penicillium implicatum, over 4-week incubation periods. The impact that such interactions have on Fusarium infection of maize grain and populations and on the production of fumonisins were all evaluated. At 0.93 and 0.95 aw, interactions of the two Fusarium spp. with other species to a large extent resulted in mutual inhibition on contact or overgrowth by the Aspergillus and Penicillium spp., regardless of temperature and growth rates of the species. Seed infection by the Fusarium spp. at 25 degrees C was influenced mainly by A. flavus and A. niger, with a reduction of infection by up to 45 and 30%, respectively, after 2 weeks. At 15 degrees C and lowered aw levels, infection was reduced, and sometimes completely inhibited, by A. niger, A. ochraceus, and P. implicatum. By contrast, populations of the Fusarium spp. (CFU per gram grain) were reduced mainly by A. flavus and P. implicatum. A. niger reduced populations of the Fusarium spp. at 0.93 and 0.95 aw, particularly after 4 weeks of incubation. Interaction with A. niger, A. ochraceus, and A. flavus at 15 and 25 degrees C and 0.98 aw resulted in a significant stimulation in fumonisin production by both Fusarium spp. Interaction with P. implicatum resulted in a decrease in fumonisin production by F. moniliforme. However, interaction with F. proliferatum resulted in an enhanced production at both temperatures and 0.95 and 0.98 aw. This study has shown that fungal interactions may act as an additional control factor in Fusarium spp. development on grain. However, although interactions lead to a decreased colonization by Fusarium, mainly at lower aw, a decrease in fumonisin production accumulation does not occur at the same time. Knowledge of these interactions is essential for improving effective control procedures for preventing growth and mycotoxin production by such fungi.

Aspergillus↗

PubMed1997

A recombinase-mediated system for elimination of antibiotic resistance gene markers from genetically engineered Bacillus thuringiensis strains.

A TnpI-mediated site-specific recombination system to construct genetically modified Bacillus thuringiensis strains was developed. Recombinant B. thuringiensis strains from which antibiotic resistance genes can be selectively eliminated were obtained in vivo with a new vector based on the specific resolution site of transposon Tn4430. For example, a cryIC gene, whose product is active against Spodoptera littoralis, was introduced into B. thuringiensis Kto harboring a cryIA(c) gene active against Ostrinia nubilalis. The resulting strain had a broader activity spectrum than that of the parental strain. It contained only B. thuringiensis DNA and was free of antibiotic resistance genes. This should facilitate regulatory approval for its development as a commercial biopesticide.

Animals↗

PubMed1996

Construction of new insecticidal Bacillus thuringiensis recombinant strains by using the sporulation non-dependent expression system of cryIIIA and a site specific recombination vector.

Bacillus thuringiensis (Bt) delta-endotoxins are safe biological insecticidal proteins whose usefulness has long been recognized. The first commercialized Bt insecticidal formulations were composed of spore-crystal preparations derived from wild-type strains. These products generally have a limited insecticidal host range and several genetically modified strains have, therefore, been constructed using transformation procedures. However, addition of a new delta-endotoxin gene to strains already harboring other delta-endotoxin genes often resulted in broader-spectrum but less potent products because they produced significantly less of each of the crystal proteins. We report expression of the coding sequence of the sporulation specific cryIC gene from the non-sporulation-dependent cryIIIA promoter. Large amounts of CryIC accumulated in various Bt strains with different genetic backgrounds. Sporulation deficient Spo0A mutants, acrystalliferous derivatives and wild-type Bt strains expressing the engineered cryIII-cryIC gene were obtained. Introduction of the cryIII-cryIC gene whose product is highly active against Spodoptera littoralis into the Kto strain harboring the cryIA(c) gene active against Ostrinia nubilalis resulted in the construction of a new strain with increased potency and broader activity spectrum than the parent strain. Large amounts of each toxin were produced and the expression of the two genes seemed to be summed, presumably because the expression systems of the two genes are different. The plasmid shuttle vector used to introduce the cryIII-cryIC gene into the different Bt hosts utilizes the specific resolution site of transposon Tn4430 to enable construction of recombinant Bt strains that are free of foreign non-Bt DNA. This should facilitate the approval and acceptance for environmental release of the insecticidal recombinant products.

Animals↗

PubMed1996

Fate of fumonisins B1 and B2 in steeped corn kernels.

Corn kernels naturally contaminated with fumonisins B1 and B2 were steeped by dipping the whole kernels in either water or a 0.3% solution of sodium bisulphite (pH = 5.7) for 6, 12, 24, 36 and 48 h. The toxin content of both the solid and the liquid were analysed for each time period. A different rate of interchange between the solid matrix and the water solution was found for FB1 and FB2 and some differences were found in the fate of both fumonisins between the two steeping systems studied. In addition, a preliminary study of the distribution of FB1 and FB2 between the germ and the remaining part of the kernel was carried out.

Carboxylic Acids↗

PubMed1996

Identification of a Bacillus thuringiensis gene that positively regulates transcription of the phosphatidylinositol-specific phospholipase C gene at the onset of the stationary phase.

A transcriptional analysis of the phosphatidylinositol-specific phospholipase C (plcA) gene of Bacillus thuringiensis indicated that its transcription was activated at the onset of the stationary phase in B. thuringiensis but was not activated in B. subtilis. The B. thuringiensis gene encoding a transcriptional activator required for plcA expression was cloned by using a B. subtilis strain carrying a chromosomal plcA'-'lacZ fusion as a heterologous host for selection. This trans activator (designated PlcR) is a protein of a calculated molecular weight of 33,762 which appears to be distantly related to PreL and NprA, regulator proteins enhancing transcription of neutral protease genes during the stationary phase of a Lactobacillus sp. and B. stearothermophilus, respectively. plcR gene transcription was analyzed in B. thuringiensis and in B. subtilis. PlcR positively regulated its own transcription at the onset of the stationary phase. There is a highly conserved DNA sequence (17 bp) 34 nucleotides upstream from the plcR transcriptional start site and 49 nucleotides upstream from the plcA transcriptional start site. As PlcR positively regulates its own transcription and plcA transcription, this conserved DNA sequence may be the specific recognition target for PlcR activation.

Amino Acid Sequence↗

PubMed1996

Water and temperature relations and microconidial germination of Fusarium moniliforme and Fusarium proliferatum from maize.

The effects of water activity (aW, 0.994-0.85 = 0.4-21.0 (-)MPa water potential), temperature (5-42 degrees C), and their interactions on microconidial germination of three isolates each of Fusarium moniliforme and Fusarium proliferatum were determined in vitro on a maize meal extract medium. Temporal germination rates of microconidia of isolates of both species were significantly influenced by both aW and temperature. Germination was very rapid at > 0.94 aW with an almost linear increase with time. Germination rates of microconidia of F. moniliforme were slower than those of F. proliferatum isolates at marginal aW levels and 5-25 degrees C, while at higher temperature (30-37 degrees C), the former germinated more rapidly than the latter. The aW minima for germination of isolates of both species was 0.88, with none occurring at 0.85 aW over a 40-day incubation period. At 37 degrees C, isolates of F. moniliforme had slightly lower aW minima than those of F. proliferatum. The narrowest range of aW for germination was at 5 degrees C, and none occurred at 42 degrees C. The effect of aW x temperature interactions on the lag phases (h) prior to germination and the germination rates (h-1) were estimated using the Gompertz model and the Zwietering equation. This showed that lag phases were shorter at 25-30 degrees C and 0.994-0.98 aW, and were increased to 10-500 h at marginal temperatures (5-10 degrees C) for F. proliferatum and longer for F. moniliforme. At marginal aW levels (0.92-0.90), lag times were increased to > 250 h. Germination rates (h-1) were different for the two species. Microconidia of F. moniliforme germinated optimally at 25-37 degrees C and 0.96-0.98 aW, but this changed to 30 degrees C at 0.90-0.94 aW, while germination of microconidia of F. proliferatum remained optimum at 30 degrees C, regardless of aW. There were statistically significant (P < 0.01) effects of aW, temperature, isolate, and two- and three-way interactions for F. proliferatum, but there were no intraisolate effects for F. moniliforme. The ecological significance of these data for understanding colonization patterns of these important fumonisin-producing fungi are discussed.

Body Water↗

PubMed1995

Fumonisins B1 and B2 and toxigenic Fusarium strains in feeds from the Spanish market.

Natural occurrence of fumonisins B1 and B2, incidence of Fusarium species, and capacity to produce fumonisins by Fusarium isolates, were investigated in 50 corn-based samples from Spain destined for animal consumption. Forty-four samples (88%) were found to be contaminated with fumonisins. The levels of contamination were very low, with a mean of 400 ng/g in the samples. We investigated the capacity of 11 isolates of Fusarium moniliforme and 19 isolates of F. proliferatum to produce fumonisins. All F. proliferatum isolates and 8 out of the 11 F. moniliforme isolates assayed produced fumonisins on a corn medium. The FB1/FB2 ratio in the isolates ranged from 1.1 to 3.5.

Animal Feed↗

PubMed1995

A survey of aflatoxins and aflatoxigenic Aspergillus flavus in corn-based products from the Spanish market.

The natural occurrence of aflatoxins B1, B2, G1 and G2, incidence of Aspergillus flavus, and capacity to produce aflatoxins by A. flavus isolates, were investigated in 100 corn-based samples from Spain destined for human and animal consumption. Only one sample for animal consumption and none for human consumption were found to be contaminated with aflatoxins B1 and B2. The levels of contamination were 0.15 micrograms/g of B1 and 0.08 micrograms/g of B2. Of 43 isolate of A. flavus, only 4 were aflatoxin producers.

Aflatoxins↗

PubMed1995

Effect of water activity and temperature on growth and fumonisin B1 and B2 production by Fusarium proliferatum and F. moniliforme on maize grain.

The effect of different water activities (aw, 0.968, 0.956, 0.944, 0.925) and temperature (25 degrees C and 30 degrees C) on colonization and production of fumonisin B1 (FB1) and B2 (FB2) on sterile layers of maize by Fusarium proliferatum and F. moniliforme isolates was determined over periods of 6 weeks. Generally, both F. moniliforme and F. proliferatum grew faster with increasing aw and best at 30 degrees C. All three isolates produced more FB1 than FB2 regardless of aw or temperature. Very little FB1 and FB2 were produced at 0.925 aw, with maximum produced at 0.956 and 0.968 aw at both temperatures tested. Most FB1 and FB2 were produced by F. moniliforme (25N), followed by F. proliferatum isolates (73N and 131N). At all aw levels and both temperatures there was an increase in FB1 and FB2 concentration with time. Statistical analyses of aw, temperature, time, two- and three-way interactions showed some significant differences between isolates and FB1 and FB2 production.

Fumonisins↗

PubMed1995

Water activity, temperature, and pH effects on growth of Fusarium moniliforme and Fusarium proliferatum isolates from maize.

The effects of water activity (aw, 0.994-0.90 identical to 0.4-14.0 (-)MPa water potential), temperature (4-45 degrees C), and pH (3.6, 5.5, 7.0), and their interactions on growth of isolates of Fusarium moniliforme and Fusarium proliferatum were determined in vitro on a maize extract agar medium. Growth of two isolates of F. moniliforme and four isolates of F. proliferatum were significantly influenced by water activity regardless of solute type used (NaCl, glycerol, or glucose). However, at steady-state aw levels, growth was optimum at 0.994-0.98 aw and reduced significantly at 0.92 aw. Further detailed studies with one isolate of F. moniliforme (25N) and two isolates of F. proliferatum (73N, 131N) showed that growth occurred over the range of 0.994-0.90 aw in the temperature range 20-35 degrees C with slight differences between species. Growth did occur at 4 degrees C and 0.994-0.96 aw, but no growth was recorded at 40 and 45 degrees C regardless of aw. Profiles of aw x temperature relations for growth of these two species were constructed from these data for the first time. Optimum pH and temperature for growth was 5.5 and 25 degrees C for both isolates of F. proliferatum, and pH 7.0 and 30 degrees C for the isolate of F. moniliforme. However, for the latter isolate at < 0.98 aw, optimum pH and temperature for growth changed. The effects of pH, temperature, and aw for single, two-way and three-way interactions were all found to be statistically significant for these three isolates. The ecological significance of this information for understanding these important fumonisin-producing fungi is discussed.

Culture Media↗