Heat resistance,biology and prevention of Diehliomyces microsporus in crops of Agaricus species

Soon after its introduction the mushroom speciesAgaricus bitorquis, which is immune to virus disease and prefers a warm climate, was threatened by the competitorDiehliomyces microsporus, false truffle. This fungus also likes warmth, and used to occur in crops ofA. bisporus.Mycelium and ascocarps were grown on several nutrient media. Optimum temperatures for mycelial growth were 26°C and 32°C, with a slight depression at 30°C. In trials in isolated growingrooms strain Somycel 2.017 ofA. bitorquis was generally used since it appeared to be highly sensitive to the competition of false truffle. Inoculation with mycelium, ascocarps or ascospores ofD. microsporus nearly always resulted in the presence of the competitor and in decreased mushroom yields. Even ten spores per m² causedD. microsporus. The time of inoculation was most important: irrespective of the kind of inoculum, inoculation only resulted in both false truffle and yeild loss, if applied from spawning until a few days after casing. Inoculation at a later date could result in false truffle, but yield was not decreased.As germination in vitro of ascospores failed, even after addition of various triggers, ascospore suspensions were treated at various temperatures for several periods. Then mushroom growing trays spawned with Somycel 2.017 were inoculated with the treated suspensions giving 7–11×10⁷ spores/m². The ascospores could not withstand 85°C for 0.5 h, 80°C for 1 h and 70°C for 3 h. Spontaneous incidence of false truffle, however, could not always be prevented and interfered with the results of these trials. It is possible that the thermal death-point of the ascospores is below 85°C. Fruiting bodies and ascospores did not survive peak-heating at the beginning and cooking out (compost temperature 12 h at 70°C) at the end of a crop. After cooking out, however,D. microsporus could still be present in the wood of trays and contaminate a following crop if no wood preservative was applied.Yield of Somycel 2.017 was reduced by the competition ofD. microsporus much more than yeilds of other strains ofA. bitorquis. The least sensitive were the highly productive strains Horst K26 and Horst K32.The effects of fungicides onD. microsporus in vitro and in growing trials did not correspond. The fungicides tested so far could not prevent or controlD. microsporus. Growing of less sensitive strains ofA. bitorquis together with sanitary measures early in the crop and at the end of the crop, however, can prevent the competitor. failure to turn up of false truffle. To understand the discrepancy between the in vitro effects of several fungicides and their effect in inoculated mushroom trays, the rate of adsorption of benomyl in the substrate and probably the interrelationships between antagonists andD. microsporus require further research. Other strains ofA. bitorquis than Somycel 2.017 appeared to be less sensitive to the competition. Among these, highly productive strains Horst K26 and Horst K32 will not be hindered byD. microsporus if the following precautions are exercised: cooking out at the end of a crop (compost temperature 70°C for 12 hours), followed by treatment of the wood with SPCP; protection by hygiene early in the crop, i.e. covering of the compost by a thin plastic sheet during mycelial growth followed by a quick execution of casing.Samenvatting De teelt van de warmteminnende champignonsoortAgaricus bitorquis, die immuun is voor virusziekte, werd al spoedig na introductie bedreigd door de eveneens warmteminnende concurrentDiehliomyces microsporus, valse truffel. Deze schimmel kwam vroeger voor in teelten vanA. bisporus; de sporen zouden een temperatuur van 82°C gedurende 5 uur kunnen overleven (Lambert, 1932). Tabel 1 geeft de myceliumgroei op verschillende voedingsbodems en de vorming van vruchtlichamen (Fig. 1A, B) weer. De optimale temperaturen voor myceliumgroei waren 26°C en 32°C, met een licht depressie bij 30°C (Fig. 2). Proeven in geïsoleerde teeltruimten werden voornamelijk uitgevoerd met Somycel 2.017, een ras vanA. bitorquis. Inoculatie met mycelium, vruchtlichamen en/of ascosporen vanD. microsporus, al of niet in reincultuur gekweekt, leidde vrijwel steeds tot de aanwezigheid van de concurrent in de geïnoculeerde teeltkisten (Fig. 1C, D), waarbij vruchtlichamen met ascosporen (Fig. 1E) gevormd werden en tot een reductie van het aantal champignons. Tien sporen per m² waren al voldoende omD. microsporus te doen aanslaan (Fig. 3). Het tijdstip van inoculatie bleek van groot belang te zijn: onafhankelijk van de aard van het inoculum leverde dit slechts zowel valse truffel als oogstreductie op, indien het werd aangebracht in de periode vanaf enten tot enkele dagen na het afdekken (Tabel 2 en Fig. 4). Inoculatie op latere tijdstippen kon wel tot valse truffel leiden, maar niet tot oogstreductie.Aangezien de kieming van ascosporen in vitro slechte resultaten opleverede, ook na toevoeging van diverse stimulantia, werden ascosporensuspensies in vitro gedurende verschillende tijden bij verschillende temperaturen behandeld; vervolgens werden teeltkisten met de behandelde suspensies geïnoculeerd (7 tot 11×10⁷ sporen/m²). De kisten waren tevoren geënt met Somycel 2.017. Een aantal proeven wees uit, dat de ascosporen 1/2 uur 85°C, 1 uur 80°C en 3 uur 70°C, niet overleefden (Tabel 3). Het spontaan optreden van valse truffel kon echter niet altijd worden voorkomen en beïnvloedde de uitkomsten van deze proeven. Daarom is het mogelijk, dat de sporen al bij een lagere temperatuur worden gedood Vruchtlichamen en ascosporen werden gedood door het uitzweten aan het begin van een teelt en door het doodstomen aan het einde van een teelt (composttemperatuur 12 uur 70°C) maar de schimmel bleek in het laatste geval wel over te kunnen blijven in het hout van teeltkisten als er vervolgens geen houtontsmettingsmiddel werd toegepast.Somycel 2.017 leed verhoudingsgewijs meer schade door concurrentie vanD. microsporus dan enkele andere rassen (Tabel 4 en. 5). Inoculatie met ascosporen bleek bij de minst gevoelige en meest produktieve rassen Horst K26 en Horst K32 slechts te gelukken in extreem droge compost; bij Somycel 2.017 daarentegen zowel in compost met een laag als met een hoog vochtgehalte. Inoculatie met mycelium veroorzaakte meer valse truffel en meer schade naarmate de compost natter was (Tabel 5).De werking van een aantal fungiciden in vitro (Tabel 6) en in teeltkisten (Tabel 7) stemde niet overeen. Aangezien de tot nu toe getoetste fungicidenD. microsporus niet kunnen voorkomen of bestrijden, moet preventie van deze concurrent worden gezocht in het telen van weinig gevoelige rassen vanA. bitorquis in combinatie met hygiënische maatregelen vroeg in en aan het eind van de teelt.     

Comparisons of frequencies and egg shell characteristics of broken and intact eggs within diverse populations of chickens

1. Differences were found among eight populations of White Plymouth Rock pullets in the frequency of broken eggs.

2. The shells of such eggs were thinner than those of intact eggs in seven of the populations.

3. The number of defective eggs, the specific gravity of the eggs, and the percentage hen‐day egg production of normal eggs were significantly correlated within populations with the incidence of broken eggs.

4. No significant correlations were found within populations between the incidence of egg breakage and either egg weight, body weight, or shank length.     

Subclinical pneumonia associated with an experimental adenovirus infection in the domestic fowl and the effect of concurrent infectious laryngotracheitis virus.

A CELO-type adenovirus (AV) isolated from fowls with respiratory disease was inoculated experimentally into the tracheas of young birds. No symptoms referable to respiratory infection were evident. Post mortem examination between days 2 and 5 after inoculation revealed pneumonia involving up to 30 per cent of the surface of the lungs. Histologically, a focal to diffuse interstitial lymphocytic infiltration and bronchiolar degeneration were present. Concurrent infections with a mild strain of infectious laryngotracheitis virus (ILT) failed to enhance the pathogenicity of either the AV or ILT infections.     

Comparative Study on Five Different Commercial Extenders for Boar Semen

Increasing interest in a longer preservation of diluted boar sperm raises questions in the field concerning the choice of the extender. The aim of this study was to evaluate the longevity of boar sperm extended in currently used commercial semen extenders. Three long-term extenders and two short-term extenders were compared for different semen quality parameters that can be assessed under routine laboratory conditions. Sperm morphology, motility, pH and bacteriological contamination were investigated during a 7-day period. The number of dead spermatozoa did not differ significantly among the extenders (p > 0.05). Sperm motility was not only related with storage period but most of all with pH, especially in long-term extenders. Differences between the different extenders were prominent (p < 0.05); the sperm preserved in only one long-term extender showed good motility during the whole test period. In all cases, the pH of the extended semen increased by 0.3-0.5 in the first days of storage and was significantly correlated with a decrease in motility. Bacteriological quality had no significant influence on motility or pH of the semen. In conclusion, we can state that in both short-term extenders and in only one long-term extender, sperm longevity, as evaluated by the parameters used in this study, was sufficient during the preservation period. To preserve the quality of diluted boar semen during long-term storage, the choice of the long-term extender is important. In addition, the monitoring of the pH of extended boar semen in our study emphasizes the importance of the buffering capacity of semen extenders.     

Viral RNA extraction using an automatic nucleic acid extractor with magnetic particles and genetic characterization of bovine viral diarrhea virus in Tokachi Province,Japan, in 2016–2017

In this study, the viral genome extraction performance of automatic nucleic acid extractors and manual nucleic acid extraction kits was compared. We showed that compared with manual kits, the automatic extractors showed superior genome extraction performance using bovine viral diarrhea virus (BVDV) genome-positive cattle sera and bovine coronavirus/infectious bovine rhinotracheitis virus-spiked cattle nasal swabs. In addition, the subgenotyping of BVDV strains detected in Tokachi Province in Japan during 2016–2017 was performed. Results showed that most of these BVDV strains belonged to subgenotype 1b, while few strains belonged to subgenotypes 1a and 2a. This study showed the high applicability of automatic nucleic acid extractors in extracting multiple viral genomes and the dominant subgenotype of BVDV in Tokachi.     

Fucose-Rich Sulfated Polysaccharides from Two Vietnamese Sea Cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera: Structures and Anticoagulant Activity

Fucosylated chondroitin sulfates (FCSs) FCS-BA and FCS-HS, as well as fucan sulfates (FSs) FS-BA-AT and FS-HS-AT were isolated from the sea cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera, respectively. Purification of the polysaccharides was carried out by anion-exchange chromatography on DEAE-Sephacel column. Structural characterization of polysaccharides was performed in terms of monosaccharide and sulfate content, as well as using a series of non-destructive NMR spectroscopic methods. Both FCSs were shown to contain a chondroitin core [→3)-β-d-GalNAc-(1→4)-β-d-GlcA-(1→]_n bearing sulfated fucosyl branches at O-3 of every GlcA residue in the chain. These fucosyl residues were different in pattern of sulfation: FCS-BA contained Fuc2S4S, Fuc3S4S and Fuc4S at a ratio of 1:8:2, while FCS-HS contained these residues at a ratio of 2:2:1. Polysaccharides differed also in content of GalNAc4S6S and GalNAc4S units, the ratios being 14:1 for FCS-BA and 4:1 for FCS-HS. Both FCSs demonstrated significant anticoagulant activity in clotting time assay and potentiated inhibition of thrombin, but not of factor Xa. FS-BA-AT was shown to be a regular linear polymer of 4-linked α-L-fucopyranose 3-sulfate, the structure being confirmed by NMR spectra of desulfated polysaccharide. In spite of considerable sulfate content, FS-BA-AT was practically devoid of anticoagulant activity. FS-HS-AT cannot be purified completely from contamination of some FCS. Its structure was tentatively represented as a mixture of chains identical with FS-BA-AT and other chains built up of randomly sulfated alternating 4- and 3-linked α-L-fucopyranose residues.     

Potential chemical time bombs in the schelde estuary

The Schelde estuary is heavily polluted with many different (micro)pollutants. This results in high concentrations in the abiotic and biotic compartments of the Schelde estuary and in various effects. the present day suboxic and anoxic conditions in the upper estuary are probably responsible for the high distribution coefficients for some trace metals in the lower Schelde estuary (Western Schelde). a future reduction in discharges with a high biological oxygen demand will therefore not only result in higher oxygen concentrations, but probably also in lower distribution coefficients for trace metals in the Western Schelde. Simulations show that lower trace metal distribution coefficients will probably result in higher concentrations of dissolved (bioavailable) trace metals, even with substantially reduced discharge rates, due to the desorption of trace metals from resuspended bottom sediments.     

Metabolism and fate of [(14)C]ethametsulfuron-methyl in rutabaga (Brassica napobrassica Mill)

The metabolism and fate of ethametsulfuron-methyl ?methyl 2-[[[[[4-ethoxy-6-(methylamino)-1,3, 5-triazin-2-yl]amino]carbonyl]amino]sulfonyl]benzoate? in rutabaga were investigated. After 72 h, absorption and translocation of [(14)C]ethametsulfuron-methyl in rutabaga did not change for the duration of the study (50 days). Less than 4% of recovered radioactivity was present in the rutabaga root. Ethametsulfuron-methyl was metabolized through a proposed unstable alpha-hydroxy ethoxy intermediate that dissipated 3 days after treatment to two major metabolites, O-desethylethametsulfuron-methyl and N-desmethyl-O-desethylethametsulfuron-methyl, as determined by liquid chromatography-mass spectrometry. It was estimated that at a spray dose of 30 g of active ingredient ha(-)(1) and a harvest weight of 0.5 kg, the edible portion of the rutabaga root would contain no ethametsulfuron-methyl and approximately 1.3 ppb total of both identified metabolites. Residue analysis and toxicological assessment show that ethametsulfuron-methyl and its metabolites should pose little or no risk to consumers of rutabagas.