Some effects of the nematodes Ditylenchus myceliophagus and Aphelenchoides composticola on the yield of the cultivated mushroom

The addition of one, ten and fifty Aphelenchoides composticola per ioo g compost at spawning reduced the total yield of sporophores by 26%, 30% and 42% respectively (P= 0·01), demonstrating a significant regression (P= 0·05) of yield on inoculum concentration. In pots inoculated at spawning with ten and fifty A. composticola per 100 g compost, cropping ceased after 12 weeks. The addition of twenty, 100 and 300 Ditylenchus myceliophagus at casing reduced the total yield of sporophores by 50%, 68% and 75 % respectively (P= 0·01). Cropping did not occur in pots inoculated at spawning, but continued for 9 weeks in pots inoculated at casing. The yield from pots inoculated at casing with either nematode was significantly greater (P= 0·05) than that from pots inoculated at spawning. When inoculated at spawning, peak populations of D. myceliophagus and A. composticola were reached 7 and 13 weeks, respectively, after spawning. Then the numbers present were unrelated to the number in the inocula.     

Comparative Assessment of biological (Nematoda: Steinernema feltiae) and chemical methods of control for the mushroom fly Lycoriella auripila (Diptera: Sciaridae)

The sciarid fly Lycoriella auripila is the major pest of mushrooms cultivated in the UK. Its larvae, which are capable of damaging the crop at all stages of production, may cause severe yield losses and can only be controlled with chemical pesticides. An indigenous isolate of the insect‐parasitic nematode Steinemema feltiae was tested as a biological control agent and its effects compared with two commonly used insecticides, diazinon and diflubenzuron. The timing of application of nematodes was found to affect their efficacy. When applied to compost during spawning, nematodes did not significantly reduce fly emergence, but they did if applied at casing when they were almost as effective as diflubenzuron. Diazinon incorporated into compost did not reduce fly emergence and was also the only treatment that did not lower the incidence of mushrooms spoiled by tunnelling of the larvae of L. auripila. When compared with untreated control plots those treated both with diazinon and diflubenzuron showed significant mean losses in yield of 10% in total weight and 17% in total numbers of mushrooms picked. In contrast, when S. feltiae was applied at casing significant mean increases in yield of 7% and 19%, respectively, were attained. Infective nematodes persisted well in casing, very few were found on sporophores.     

The Development of a Bioluminescence Assay to Compare the Efficacy of Biocides Incorporated into Plasticised PVC

A bioluminescence assay was developed using the expression of the luxAB genes in Pseudomonas veronii to allow the efficacy of biocides incorporated into plasticised polyvinylchloride (pPVC) to be determined in situ. A maximum number of cells was found to adhere to the surface after 18 h as measured by bioluminescence, radiolabelling and viable cell counts. A positive correlation was found between the level of bioluminescence and numbers of viable cells attached to the pPVC. When the biocide 10, 10-oxybisphenoxyarsine (OBPA) was incorporated into the pPVC, both bioluminescence and viable cell number were reduced by ca 60% at a concentration of 750 ppm and by >99% at 2250 ppm. When the biocide 2,3,5,6-tetrachloro-4-(methylsulphonyl)pyridine (TCMP) was incorporated into the pPVC, no reduction in viability or bioluminescence was seen after 18 h. However, over a period of 72 h at a concentration of 2250 ppm TCMP, both viable cell number and bioluminescence decreased steadily after 36 h until after 72 h, both bioluminescence and viable cell counts were less than 1% of the initial values. The viability of attached cells can therefore be measured in situ in a sensitive real-time assay by measuring bioluminescence allowing the efficacy of biocides incorporated into plastics to be compared.     

Comparative effects of three insect growth regulator insecticides and a dipteran-active strain of Bacillus thuringiensis on cropping of the cultivated mushroom Agaricus bisporus

Three insect growth regulator insecticides and an entomopathogenic strain of Bacillus thuringiensis (GC327), products effective against the mushroom sciarid, Lycoriella auripila, were compared for their effect on mushroom cropping. Cyromazine and diflubenzuron were applied as a surface drench to mushroom compost before or after pasteurisation (at filling or spawning, respectively); admixed into casing material (at casing); or at a combination of these times. Hexaflumuron and GC327 were applied only at filling and casing, respectively. The presence of the target pest, L. auripila, had no effect on treatment trends, although it was accounted for in the analysis by use of a yield model. The trial was notable for the disparate effects that cyromazine and diflubenzuron casing treatments had on mushroom cropping. Cyromazine treatments that included application at casing resulted in increases in yield, compared to the untreated control whereas, with diflubenzuron, the opposite was true, with treatment at casing alone causing the greatest reduction overall (10%). GC327 applied at casing was also conspicuous for giving a 13% increase in yield. Treating the crop at casing with either cyromazine or GC327, therefore, resulted in a 15% or 24% increase in yield, respectively, compared to a similar treatment with diflubenzuron. Hexaflumuron applied at filling caused increases in yield compared to application of cyromazine at filling and cyromazine or diflubenzuron at spawning. There were also effects on crop timing. The addition of a cyromazine casing treatment normally caused the distinct flushes of mushrooms to be produced significantly earlier than the untreated control (up to 2.5 days), as did GC327. With diflubenzuron, the earlier flushes were only produced by those treatments that did not include a casing application. The combinations that included a casing treatment with diflubenzuron initially produced mushroom flushes earlier than the untreated control. They became either synchronous with the control or they were delayed. From the crop tolerance perspective, therefore, cyromazine and GC327 would be the sciarid control products of choice for a commercial mushroom grower.     

Cometabolic degradation of 4-chlorophenol by Alcaligenes eutrophus

 Alcaligenes eutrophus was grown in batch cultures using either phenol as a sole substrate or mixtures of phenol and 4-chlorophenol. Phenol was found to be the sole source for carbon and energy while 4-chlorophenol was utilized only as a cometabolite. Maximum growth rates on phenol reached only 0.26 h^-1, significantly below the growth rates reported earlier with Pseudomonas putida. The cometabolite was found to decrease biomass yield and increase lag time before logarithmic growth occurred. Both phenol and 4-chlorophenol were found to inhibit the growth rate linearly with maximum concentrations of 1080 ppm and 69 ppm respectively, beyond which no growth occurred. The best-fit parameters are incorporated into a simple, dynamic (i.e. time-varying) model capable of predicting all the batch growth conditions presented here. It is shown that P. putida is capable of faster bioremediation when phenol is the sole carbon source or for mixed substrates with low concentrations of the cometabolite, but for high concentrations of 4-chlorophenol, A. eutrophus becomes superior because of the long lag times that occur in the Pseudomonas species. Received: 25 January 1996/Received revision: 13 March 1996/Accepted: 15 April 1996     

Effect of irrigation treatments upon the chemical constituents ofHibiscus sabdariffa L.

Summary The effects of Se and P on the dry matter yield and chemical composition of Raya (Brassica juncea Cos. var. R.L. 18) were studied in the green-house. The dry matter yield in P treated and untreated pots increased with Se application upto 2.5 ppm. With further increase in Se dose upto 10 ppm, dry matter yield decreased. The increase in P dose from 0 to 50 ppm increased dry matter yield in all Se treated and untreated pots but 100 ppm P rather showed decrease in dry matter yield as compared to 50 ppm P.The Se concentration increased by about 100 fold with the application of 10 ppm Se over 0 ppm Se where P was not added whereas increase was 258 and 336 times when 50 ppm and 100 ppm P was added, respectively. Inorganic, organic and total P increased with increasing Se and P. The increase in inorganic P was more than organic and total P. S and N concentration decreased with increasing Se application and increased with P application.Crude protein showed the same behaviour as N. The total sulphur-containing amino acids (which ranged from 39 to 49% of crude protein) and individual sulphur-containing amino acids like methionine (16.9 tot 20%) cysteine (9.8 to 13%) and cystine (12.3 to 15.9%) decreased with the increasing selenium but increased with increasing phosphorus. N/S ratio in the plant showed significant negative correlation with total sulphur-containing amino acids (r=–0.940^**), methionine (r=–0.951^**), cysteine (r=–0.929^**) and cystine (r=–0.920^*) whereas total sulphur in the plant showed positive significant correlation with methionine (r=0.805^**), cysteine (r=0.924^**) and cystine (r=0.821^**).     

Insecticide granule distribution and the control of Lycoriella auripila (Diptera: Lycoriidae) in mushroom casing material

In laboratory tests the distribution of insecticide in peat–chalk mushroom casing greatly affected the number of adult sciarids which emerged from larvae placed in 50 g samples. Only thionazin markedly reduced emergences when granules were clumped in the centre of the sample of casing. The slope of dosage mortality data for clumped granule treatments was less steep than for mixtures and gave estimated LD₉₅ values of 16300 and 26.8 ppm respectively. Thus, the number of adults which emerged when granules amounting to 6.4 mg of chlorfenvinphos were clumped was 65% of those emerging when only 0.4 mg a.i. was mixed into 50 g casing. However, adults did not emerge when 3.2 mg of the same insecticide was mixed throughout the sample. Increased doses did not, therefore, compensate for inadequate mixing. With 0.2–0.4 mg a.i. chlorfenvinphos mixed through the casing, about three times as many sciarids emerged with the intact granules than when pulverized granules were used. This showed the lack of mobility of this chemical in peat-chalk mixtures and the need for it to be homogeneously incorporated in commercial casing. Pirimiphos-ethyl, diazinon and thionazin also gave improved kills when mixed into the casing rather than clumped. A χ² test of the mortality data suggested a non-linear dosage-mortality relationship for treatments amounting to half the dish. However, LD₉₅ estimates of 37.5 and 62.2 ppm were obtained for mixed and quadrant treatments. Thus, when 3.2 mg chlorfenvinphos was mixed into only half of the volume of casing, the number of adults emerging was only 6% of that from untreated casing. However, larval movement was negligible in untreated casing and this appeared to limit the effectiveness of chlorfenvinphos and pirimiphos-ethyl when they were not evenly distributed throughout the casing. With thionazin and diazinon, mobility of the insecticide appeared to compensate for uneven distribution.     

Influence of plant growth regulators on growth, morpho-physiological characters and yield of summer sesame (Sesamum indicum L. cv. Rama) under moisture stress

Field experiments were conducted with sesame (Sesamum indicum L. cv. Rama) for two years (1997 and 1998) to study the effect of three level of irrigation (F+C, B+C, B+F+C) and two growth regulators (CCC, 200 ppm CCC; 100 ppm and BX-112, 100 ppm; BX-112, 50 ppm) on growth (root and shoot length, average number of primary branches/plant), morpho-physiological growth parameters(LAI, LAD, CGR and NAR), yield attributing parameters(average number of capsule/plant, average number of seeds/capsule) and seed yield. Irrigation at B+F+C stage showed significant effect on these parameters. Among the growth regulators, CCC, 200 ppm showed remarkable results on these parameters and seed yield. Seed yield in CCC, 200 ppm treatment was more than 53% in comparison to water soaked seeds. The interaction between irrigation and PGR showed better seed yield and it was concluded that the growth regulator CCC might be utilized for enhancement of seed yield of summer sesame under field condition.     

In Vitro Sensitivity of <Emphasis Type="Italic">Rhizobium</Emphasis> and Phosphate Solubilising Bacteria to Herbicides

Nitrogen fixing bacteria, rhizobia and phosphate solubilizing bacteria (PSB) are the commonly applied microbial inoculants in grain legumes (Pulses). It is important to apply herbicides to control weeds in order to augment yield of the crop. The herbicides may however, be incompatible with the microbial inoculants. This study compared the effect of the recommended pre-plant incorporated herbicide, fluchloralin (20.25 × 10⁴ ppm) and pre-emergence herbicide, pendimethalin in two doses (9 × 10⁴ and 15 × 10⁴ ppm) on the growth and survival of mungbean Rhizobium and PSB, under laboratory conditions. These herbicides were also used under field conditions in conjunction with biofertilizers (R, PSB) to improve grain yield of mungbean. It was found that fluchloralin (20.25 × 10⁴ ppm) and the lower dose of pendimethalin (9 × 10⁴ ppm) had no adverse effect on growth of Rhizobium and PSB. The higher dose of pendimethalin (15 × 10⁴ ppm) was safe on PSB but it imposed a retarding effect on the growth of Rhizobium.     

Barium variation in Acropora palmata and Montastrea annularis

The barium contents of Acropora palmata and Montastrea annularis are distinct both in their averages (6 and 9 ppm, respectively) and, more impressively, in their variability (5 to 7 ppm and 6 to 15 ppm). A. palmata contained about 50% more Ba²⁺ than M. annularis and exhibited more than three times as much variation, as measured by their respective coefficients of variation. In contrast, the means and coefficients of variation of Sr²⁺ in these groups differed by only 6% and 15%. No obvious environmental or post-depositional causes for the inter-and intra-specific variation of Ba²⁺ could be found. Previous experiments indicate that the partitioning of Ba²⁺ into aragonite depends directly on the rate of precipitation. This suggests that Ba²⁺ does not substitute for Ca²⁺ but instead is incorporated by occlusion. Since growth rates in A. palmata often exceed those of M. annularis, this appears to be the mechanism which generates higher and more variable concentrations of Ba²⁺ in A. palmata.     

Incidence,identification and pathogenicity of Cladobotryum mycophilum,causal agent of cobweb disease on Agaricus bisporus mushroom crops in Spain

Between 2008 and 2011, outbreaks of cobweb were observed in commercial white button mushroom (Agaricus bisporus) crops in Castilla‐La Mancha (Spain). In the last 3 years, the presence of the disease has notably increased resulting in serious economic losses. Based on morphological and genetic analyses, the casual agent of cobweb was identified as Cladobotryum mycophilum. A. bisporus mushroom crops were surveyed over a 2‐year period to estimate the incidence of cobweb. The presence of the disease was detected in 32% of the mushroom crops observed, being of particular concern in autumn (44% of crops infected) and winter (37%). As regards the casing material, the percentage of crops affected by cobweb was 34% in crops using mineral casing and 29% in those cased with a peat‐based casing, with no statistical relationship between the casing and the presence of cobweb. Two cropping trials inoculated with C. mycophilum were set up to evaluate the pathogenicity of the causal agent of cobweb in three peat‐based casings (C1, C2 and C3). The effect of cobweb on mushroom productivity was evaluated by comparing mushroom production and the cobweb patches detected in the casing soil. The decrease in total yield of mushrooms attributed to cobweb reached 12.9% with C2, and the crop area colonised by cobweb reached a final percentage of 36% with C3.     

The incidence of benomyl tolerance in Verticillium fungicola, Mycogone perniciosa and Hypomyces rosellus in mushroom crops

Of the 229 isolates of Verticillium fungicola examined 63 % were found to show some benomyl tolerance and 53% were very tolerant having EDg, values in excess of 50 ppm. There was a tendency for the tolerant isolates to have a slower growth rate than the sensitive ones on unamended potato dextrose agar (PDA) and forty-five isolates (19 %) showed growth stimulation at 5 ppm benomyl. No major differences were found in ten isolates of V. fungicola tested on benomyl, thiabendazole and thiophanate methyl agars or of twelve isolates compared on benomyl and carbendazim hydrochloride agars. Tolerant isolates of V. fungicola and higher levels of crop loss were linked in the east and north of the country but not in the south-east. Tolerance was not found in sixty-six isolates of Mycogone perniciosa or twenty-four of Hypomyces rosellus. Benomyl was used on 89 % of the farms in the survey either applied to the casing (34%) or as a spray (55 %) or both (10%).     

Biological activities of extracts from traditionally used Nigerian plants against the European corn borer, Ostrinia nubilalis

Preliminary investigations with ethanolic (EtOH) extracts from five Nigerian plants show that extracts of Piper guineense Schum and Thonn (Piperaceae), Cedrela odorata L. (Meliaceae), Dennettia tripetala G. Baker (Annonaceae) and Aframomum melegueta (Rosch) K. Schum (Zingiberaceae) in artificial diets significantly reduced larval growth of European corn borer (ECB), Ostrinia nubilalis Hubner, at a concentration of 1000 ppm (0.1%). An extract of Xylopia aethiopica (Dunal) A. Rich (Annonaceae) was ineffective. When the extracts were subsequently incorporated into artificial diets at 300 ppm and offered to neaonates, larval mortality increased in the order A. melegueta (13%), D. tripetala (13%), P. guineense (27%), and C. odorata (48%). Larval and adult emergence periods increased with increasing concentration of P. guineense, C. odorata and D. tripetala indicating a toxic response. Nutritional indices for habituated third instar larvae with the two most promising plant extracts, P. guineense and C. odorata, showed that the efficiencies of conversion of digested food (ECD) was significantly reduced at 300 ppm suggesting a postdigestive toxicity of the extracts. P. guineense and C. odorata extracts show the best potential for development as botanical insecticides.     

Behaviour of Candida utilis during growth in relation to Mg2+ and K+ concentration in sugar cane molasses

Magnesium and potassium ions are present in sugar cane molasses in a concentration of about 0.3% and 2.5%, respectively, which is high enough to support biomass production from Candida utilis. Culture broth with 40 g/l of total reducing substances supplemented by the addition of 1 ppm of Mg²⁺ leads to a higher yield (Y x/s) in batch fermentation experiments. The subsequent addition of Mg²⁺ up to 10 ppm decreases the yield coefficient from 0.53 to 0.42 without affecting the growth rate. Fermentation media supplemented with 1 to 10 ppm of K⁺ decreased both the yield coefficient and the specific growth rate. A Mg/K ratio of about 0.1 seems to be optimal for yeast biomass propagation.     

Biology and control of the sciarid Lycoriella auripila Winn. (Diptera: Lycoriidae) in mushroom culture

The life-history of L. auripila, as a pest of mushrooms, is outlined and an explanation is given of the different types of damage caused. The most satisfactory control is based on the incorporation of insecticides in compost at spawning. Emulsions of chlorfenvinphos and diazinon at 50 ppm are lethal to young larvae for at least 7 weeks but granular formulations of the former must be applied at 100 ppm to achieve comparable results. Drenches of 0–01% malathion, applied at about 40 gal/1000 ft², are effective against larvae in the casing.     

13C CP/MAS NMR Spectral Analysis of 6-O-Tosyl, 6-Deoxy-6-iodo,and 6-Deoxy Derivatives of N-Acetylchitosan in a Solid State

6-O-Tosyl (1, d.s. 0.94, 80% yield), 6-deoxy-6-iodo (2, d.s. 0.49, 86% yield) and 6-deoxy (3, d.s. 0.49, 50% yield) derivatives of N-acetylchitosan were prepared, and a ¹³C CP/MAS NMR spectral analysis was performed because no suitable solvent for 3 was found. The ¹³C signal for CH₃ at C-6 in 3 was detected at 18.9 ppm, and that for C-4 in 1–3 appeared at 72.2–72.7 ppm, which is in a higher magnetic field than those (82.5–86.0 ppm) in N-acetylchitosan, 6-O- (ethylthio), 6-O-(benzylthio)- and 6-O-(methylthio)-thiocarbonyl derivatives, chitosan, and chitin. This strongly suggests a different molecular conformation for 1–3.     

Effects of bendiocarb and diflubenzuron on mushroom cropping

When mixed into the casing or compost layers of a mushroom bed in the absence of pests, bendiocarb decreased yield and number of mushrooms according to concentration. The most severe effects were on mushroom number at the two highest rates used (100 and 1000 μg/g), and there were large increases in mushroom size. Effects of bendiocarb incorporation in the compost diminished with time, and there was partial compensation in yield and numbers at the fourth flush. The action of bendiocarb persisted when it was mixed into the casing. Diflubenzuron showed some opposite effects at lower concentrations. When either mixed into, or drenched onto the casing at the commercial rate (30 μg/g), yield and size were both increased and the timing of the flushes was unaffected. At the two higher concentrations (180 and 1080 μg/g), reductions in yield and number and an increase in mushroom size were shown. However, these effects became more severe with time, especially those on mushroom number, possibly due to the accumulation of a toxic breakdown product.     

Relative contribution of nematodes (Caenorhabditis elegans) and bacteria towards the disruption of flushing patterns and losses in yield and quality of mushrooms (Agaricus bisporus)

Laboratory tests of bacteria isolated from the body surface, or from the gut, of a saprophagous rhabditid nematode Caenorhabditis elegans infesting mushrooms (Agaricus bisporus) showed that some bacteria enhanced nematode reproduction and that others inhibited it. As some bacteria were shown to inhibit mycelial growth of the mushroom, the effects of Acinetobacter calcoaceticus var. anitratus, Enterobacter cloacae and Serratia liquefaciens, either alone or in combination with C. elegans, on the flushing patterns, quality and yield of A. bisporus (strain Horst U3) were studied. Bacteria alone had little effect on flushing patterns whereas C. elegans delayed the onset of mushroom production and significantly disrupted the growth pattern of crops, with mushrooms appearing more regularly and not within obvious flushes. Inoculation with bacteria resulted in ‘browning’ of mushrooms that was even more pronounced in C. elegans treatments. Characteristic distortion of sporophores was observed only in the presence of C. elegans. Nematodes commonly colonised sporophores. Bacteria affected the size of nematode populations both on the sporophores and in the casing. Significant yield loss occurred; up to 10% when bacteria were inoculated, up to 27.8% when C. elegans was inoculated, and up to 35% with both bacteria and nematodes. Synergism between C. elegans and A. calcoaceticus var. anitratus was observed; the combination resulted in significantly greater reduction in mushroom yield than any other treatment. It is concluded that bacteria contribute to yield loss and quality deterioration in A. bisporus but that the effects are far greater in the presence of C. elegans.     

Effects of Caenorhabditis elegans (Nematoda: Rhabditidae) on yield and quality of the cultivated mushroom Agaricus bisporus

Thirteen species of saprobic rhabditid nematodes (11 genera) were identified from samples of compost and casing material collected from mushroom farms in the British Isles. Caenorhabditis elegans, the most frequently found saprobe, was mass-produced monoxenically and its effects on the cultivated mushroom, Agaricus bisporus (strain U3) were studied. C. elegans did not multiply in well-prepared, pasteurised, spawned compost, whereas casing material proved to be a highly suitable environment for its reproduction. An initial casing inoculum of 10⁶ nematodes/crate of compost (7.5 kg), caused a significant reduction in mushroom yield. Losses in total mushroom yields of 11%, 20% and 26% were caused by initial inoculum rates of 10⁶, 10⁷and 2 × 10⁷ nematodes/crate, respectively. Yields were negatively correlated with the initial nematode inoculation level and regression equations were derived. The nematode treatments caused fewer mushrooms to be produced and an absence of the usual distinctive flushing patterns. C. elegans caused considerable deterioration in mushroom quality and characteristic distortion of mushrooms. Individual sporophores were mis-shapen, notched and had brown or violet coloured grills. Up to 3.8%, 6.7% and 10.8% of total weight and 3.5%, 5.4% and 8% of total numbers of mushrooms were distorted at the three highest nematode inoculum rates tested. Weights and numbers of distorted mushrooms were positively correlated with the initial nematode population. C. elegans commonly colonised sporophores.     

PHYTOCHEMICAL STUDIES IN THE FAMILY MALVACEAE. I. COMPARISON OF ESSENTIAL OILS OF SIX SPECIES BY GAS-LIQUID CHROMATOGRAPHY

When the chemical constitution of the essential oils of six species within three genera of the family Malvaceae were investigated, the essential oils of Gossypium hirsutum L. and Cienfuegosia rosei Fryxell were found to contain 67.0 % and 58.2 % hydrocarbons, respectively. Those of Hibiscus lasiocarpus (Cav.) and Callirhoe involucrata (Nutt.) Gray contained equal percentages of hydrocarbons (14 %), but that of Cienfuegosia drummondii (A. Gray) Lewt. contained only 2.9 % hydrocarbons. Beta-bisabolol was the major oxygenated component in G. hirsutum (13.7 %), Hibiscus militaris (Cav.) (55.0 %), and Hibiscus lasiocarpus (32.6 %), but the major constituents in the essential oil of C. involucrata were myrtenal (32.0 %) and trans-2-cis-6-nonadienal (12.0 %), and the major oxygenated component in C. rosei was an unknown sesquiterpene alcohol (17.4 %) with chemical and physical properties resembling those of beta-bisabolol. The concentration of total oil was 150.0 ppm in G. hirsutum; 50.0 ppm in H. militaris; 46.3 ppm in C. rosei; 41.0 ppm in H. lasiocarpus; 25. 4 ppm in C. drummondii; and 10.0 ppm in C. involucrata, all based on wet weight.