Puffball Research Articles

Molecular and morphological analyses confirm Rhizopogon verii as a widely distributed ectomycorrhizal false truffle in Europe, and its presence in South America.

The genus Rhizopogon includes species with hypogeous or subepigeus habit, forming ectomycorrhizae with naturally occurring or planted pines (Pinaceae). Species of the genus Rhizopogon can be distinguished easily from the other hypogeous basidiomycetes by their lacunose gleba without columella and their smooth elliptical spores; however, the limit between species is not always easy to establish. Rhizopogon luteolus, the type species of the genus, has been considered one of the species that are more abundant in Europe, as well as it has been cited in pine plantation of North and South America, different parts of Africa, Australia, and New Zealand. However, in this study, based on molecular analyses of the ITS nuclear ribosomal DNA (nrDNA) sequences (19 new sequences; 37 sequences from GenBank/UNITE, including those from type specimens), we prove that many GenBank sequences under R. luteolus were misidentified and correspond to Rhizopogon verii, a species described from Tunisia. Also, we confirm that basidiomes and ectomycorrhizae recently collected in Germany under Pinus sylvestris, as well as specimens from South of Brazil under Pinus taeda belong to R. verii. Thanks to the numerous ectomycorrhizal tips collected in Germany, a complete description of R. verii/P. sylvestris ectomycorrhiza is provided. Moreover, since in this paper the presence of R. verii in South America is here reported for the first time, a short description of basidiomes collected in Brazil, compared with collections located in different European herbaria, is included.

Enrichment of heavy metals in coal gangue by puff balls and mechanism research

Coal gangue with/without wild puff balls growing on or covering up were analyzed by means of inductively coupled plasma atomic emission spectrometry (ICP-AES) and atomic fluorescence spectrometry (AFS) to detect the contents of heavy metal elements in the plants. Seven heavy metals were chosen to do the experiments, which showed that the puff balls have strong enrichment capacity for heavy metals (As, Pb, Mn and Ni). The highest enrichment level was Mn, 220.3 mg/kg (dry weight) for puffball fruiting bodies, and the highest As, Cr and Pb enrichment levels for puffball mycelium were 111, 265.3 and 86 mg/kg, respectively. Cd, Mn and Ni could transfer from puffball mycelium to its fruiting bodies, while As, Cr, Pb and Co mainly accumulated in the mycelium. The contents of five organic acids (tartaric acid, glycolic acid, lactic acid, citric acid and malic acid) in puffballs were determined with high performance liquid chromatography. With water, ethanol and ethyl acetate as solvents, the puffball extracts performed dissolution reactions with coal gangue, in which the water extract (containing a large amount of organic acids) was most effective in releasing heavy metals, especially As, Pb, Cr, Mn, Co and Ni from coal gangue. As a pioneer species, puff balls absorbed the hazardous trace elements in coal gangue matrix to reduce its eco-toxicity, on the other hand, it raised the pH value of coal gangue, improving the vegetation growing conditions for coal gangue matrix to a certain extent and laying the foundation for ecological restoration of coal gangue dumps.

Studies on volatile organic compounds of some truffles and false truffles

Results of solid phase micro-extraction coupled to gas chromatography and mass spectrometry analyses, accomplished on sporophores of 11 species of truffles and false truffles, are reported. Volatile organic compounds (VOCs) found in Gautieria morchelliformis were dimethyl sulphide, 1,3-octadiene, 3,7-dimethyl-1,6-octadien-3-ol, amorphadiene, isoledene and cis-muurola-3,5-diene. In Hymenogaster luteus var. luteus, presence of 1,3-octadiene, 1-octen-3-ol, 3-octanone, 3-octanol and 4-acetylanisole was revealed. Two VOCs, 4-acetylanisole and β-farnesene, constituted aroma of Hymenogaster olivaceus.Melanogaster broomeanus exhibited as components of its aroma 2-methyl-1,3-butadiene, 2-methylpropanal, 2-methylpropanol, isobutyl acetate, 3,7-dimethyl-1,6-octadien-3-ol, 3-octanone and β-curcumene. VOC profile of Octavianina asterosperma was characterised by the presence of dimethyl sulphide, ethyl 2-methylpropanoate, methyl 2-methylbutanoate and 3-octanone. Tuber rufum var. rufum and Pachyphloeus conglomeratus showed the presence of dimethyl sulphide only.

Torrecillasite, Na(As,Sb)43+O6Cl, a new mineral from the Torrecillas mine, Iquique Province, Chile: description and crystal structure

AbstractThe new mineral torrecillasite (IMA2013-112), Na(As,Sb)43+O6Cl, was found at the Torrecillas mine, Iquique Province, Chile, where it occurs as a secondary alteration phase in association with anhydrite, cinnabar, gypsum, halite, lavendulan, magnesiokoritnigite, marcasite, quartz, pyrite, scorodite, wendwilsonite and other potentially new As-bearing minerals. Torrecillasite occurs as thin colourless prisms up to 0.4 mm long in jack-straw aggregates, as very thin fibres in puff balls and as massive intergrowths of needles. Prisms are elongated on [100] with diamond-shaped cross-section and irregular terminations. Crystals are transparent, with adamantine lustre and white streak. The Mohs hardness is 2½, tenacity is brittle and fracture is irregular. Cleavage on (001) is likely. The calculated density is 4.056 g cm−3. Optically, torrecillasite is biaxial (−) with α = 1.800(5), β = 1.96(1), γ = 2.03(calc.) (measured in white light). The measured 2V is 62.1(5)°, no dispersion or pleochroism were observed, the optical orientation isX=c,Y=b,Z=a. The mineral is very slowly soluble in H2O, slowly soluble in dilute HCl and rapidly soluble in concentrated HCl. The empirical formula, determined from electron-microprobe analyses, is (Na1.03Mg0.02)∑1.05(As3.39Sb0.62)∑4.01O6.07Cl0.93. Torrecillasite is orthorhombic,Pmcn, a= 5.2580(9),b= 8.0620(13),c= 18.654(3) Å,V= 790.7(2) Å3andZ= 4. The eight strongest X-ray powder diffraction lines are [dobsÅ(I)(hkl)]: 4.298(33)(111), 4.031(78)(014,020), 3.035(100)(024,122), 2.853(39)(115,123), 2.642(84)(124,200), 2.426(34)(125), 1.8963(32)(225) and 1.8026(29)(0·1·10,233). The structure, refined toR1= 4.06% for 814Fo>4σFreflections, contains a neutral, wavy As2O3layer parallel to (001) consisting of As3+O3pyramids that share O atoms to form six-membered rings. Successive layers are flipped relative to one another and successive interlayer regions contain alternately either Na or Cl atoms. Torrecillasite is isostructural with synthetic orthorhombic NaAs4O6Br.

Free amino acid profiling in the giant puffball mushroom (Calvatia gigantea) using UPLC–MS/MS

Wild edible and medicinal mushroom, Calvatia gigantea, was quantitatively analyzed for the determination of its free amino acids using ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). The concentrations of total free amino acids, essential and non-essential amino acids were 199.65mg/100g, 113.69mg/100g, and 85.96mg/100g in C. gigantea, respectively. This study showed that C. gigantea, so called a giant puffball mushroom, has free amino acids content. The essential amino acids: tryptophan, isoleucine, valine, phenylalanine, leucine, threonine, lysine, histidine, methionine, and the non-essential amino acids: tyrosine, 4-hyrdroxy proline, arginine, proline, glycine, serine, alanine, glutamine, glutamic acid, aspargine, aspartic acid were detected.

Two Consecutive Clusters of Toxic Anterior Segment Syndrome

To present clinical findings and etiologic investigation of two consecutive clusters of toxic anterior segment syndrome (TASS) after uncomplicated phacoemulsification cataract surgery. At the Veterans Affairs Northern Indiana Health Care System Fort Wayne campus, 11 consecutive patients on two separate days in 2011 underwent clear corneal incision phacoemulsification cataract surgery by the same surgeon. On the first postoperative day, all patients had 1+ to 2+ diffuse limbus to limbus stromal edema and 2+ to 4+ anterior chamber white blood cells. Five eyes had inflammatory plaques on the surface of the intraocular implant, six had fibrin, three had hypopyon, and one had inflammatory debris "puff ball." Visual acuity ranged 20/70 to 20/400. Treatment included moxifloxacin four times a day, diclofenac four times a day, and prednisolone acetate 1% every 1 to 2 hours. In all patients, active inflammation and corneal edema resolved within 6 weeks, and visual outcome was 20/20. Cataract surgery was paused for 5 months after the first cluster of TASS and then immediately paused again after restarting because of a second cluster. Although no specific causes were identified, etiologic investigation resulted in the implementation of multiple changes. The changes included new replacement surgical instruments, disposable irrigation/aspirator tip and handle, risk of residue on reusable instruments minimized, preservative-free medications used when available, ophthalmic ointment eliminated, manufacturers' recommendations followed exactly, and eye instruments processed separately. Toxic anterior segment syndrome did not occur when surgery resumed 11 months after the last cluster. There are multiple possible etiologies of TASS. However, as in our clusters, specific causes are often not identified. Thorough review of all steps in surgery, processing of equipment and preparation of injectable solutions and materials, and adoption of best practices can prevent additional cases of TASS. Prompt diagnosis and treatment of TASS are extremely important because this leads to a desirable outcome.

Chemical constituents and antioxidant activity of the Arctic mushroomLycoperdon mollePers

The biochemical adaptations of fungi to the harsh conditions of the Arctic may mean that these organisms have properties useful to people. Using samples of the puffball mushroom Lycoperdon molle Pers. (Basidiomycota, Fungi) collected at Ny-Ålesund, Svalbard, we examined the in vitro antioxidant potential of this species by investigating its free-radical scavenging (FRS) activity, inhibition of lipid peroxidation (ILP) and Trolox equivalent antioxidant capacity (TEAC). The FRS activity of the samples in various organic solvents, including methanol, ethanol, acetone and dimethyl sulfoxide (DMSO), were found to be in the range of 44.00-89.60%, while ILP activities ranged from 32.00 to 54.41%. The methanol extract showed the highest levels of FRS (89.60%) and ILP (54.41%) compared to standard antioxidants butylated hydroxyanisol and butylated hydroxytoluene (BHT). The TEAC value was also found to be higher compared to the standard water soluble vitamin E analogue Trolox (3.9 mM). Antimicrobial screening of Lycoperdon molle extracts was negative to the tested microorganisms. Using electrospray ionization mass spectrometry (ESI-MS), we determined that the samples contained compounds such as phosphoethanolamine, monomethyl arsenic acid, phosphatidyl glycerol, phosphoionositol, phosphoserine and lysophosphatidyl choline. We found that Lycoperdon molle showed strong antioxidant abilities compared to the standards, suggesting that this and perhaps other Arctic mushrooms could be valuable sources of natural antioxidants for the pharmaceutical industry. To our knowledge, this is the first report of the antioxidant activity in any Arctic mushroom.Keywords: Lycoperdon; fungi; antioxidative potential; HPLC; ESI-MS; Arctic(Published: 12 March 2012)Citation: Polar Research 2012, 31, 17329, DOI: 10.3402/polar.v31i0.17329

Species delimitation in taxonomically difficult fungi: the case of Hymenogaster.

BackgroundFalse truffles are ecologically important as mycorrhizal partners of trees and evolutionarily highly interesting as the result of a shift from epigeous mushroom-like to underground fruiting bodies. Since its first description by Vittadini in 1831, inappropriate species concepts in the highly diverse false truffle genus Hymenogaster has led to continued confusion, caused by a large variety of prevailing taxonomical opinions.MethodologyIn this study, we reconsidered the species delimitations in Hymenogaster based on a comprehensive collection of Central European taxa comprising more than 140 fruiting bodies from 20 years of field work. The ITS rDNA sequence dataset was subjected to phylogenetic analysis as well as clustering optimization using OPTSIL software.ConclusionsAmong distinct species concepts from the literature used to create reference partitions for clustering optimization, the broadest concept resulted in the highest agreement with the ITS data. Our results indicate a highly variable morphology of H. citrinus and H. griseus, most likely linked to environmental influences on the phenology (maturity, habitat, soil type and growing season). In particular, taxa described in the 19th century frequently appear as conspecific. Conversely, H. niveus appears as species complex comprising seven cryptic species with almost identical macro- and micromorphology. H. intermedius and H. huthii are described as novel species, each of which with a distinct morphology intermediate between two species complexes. A revised taxonomy for one of the most taxonomically difficult genera of Basidiomycetes is proposed, including an updated identification key. The (semi-)automated selection among species concepts used here is of importance for the revision of taxonomically problematic organism groups in general.

Lycoperdonosis in Two Dogs

Lycoperdonosis is a rare respiratory disease that results from the inhalation of spores released from the Lycoperdon (puffball) mushroom. In the present study, 2 cases of confirmed canine lycoperdonosis are described. The first case presented to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania, and the second case was submitted for postmortem examination to the University of Tennessee Veterinary Teaching Hospital. Both dogs presented in respiratory distress, and owners reported that the dogs had been playing or digging in areas with puffball mushrooms prior to the onset of clinical signs. In the initial case, thoracic radiographs revealed a diffuse interstitial and multifocal alveolar pulmonary pattern. Despite aggressive medical treatment and mechanical ventilation, the dog continued to worsen and was euthanized. Postmortem examination revealed firm lung lobes and enlarged tracheobronchial lymph nodes. Histologically, there was a severe diffuse histiocytic and pyogranulomatous bronchointerstitial pneumonia. Throughout the lung and lymph nodes, most commonly within macrophages, were round, 3-5µm in diameter, Gomori methenamine silver-positive structures, consistent with Lycoperdon spores. An approximately 750-base pair DNA fragment was amplified from lung of both cases by polymerase chain reaction using primers specific to yeast ribosomal DNA, and the sequence of the fragment was determined to be most closely related to Lycoperdon pyriforme. Importantly, reexamination of an endotracheal wash from the initial case revealed intrahistiocytic spores, suggesting that airway sampling may assist in diagnosing lycoperdonosis.

The neglected hypogeous fungus Hydnotrya bailii Soehner (1959) is a widespread sister taxon of Hydnotrya tulasnei (Berk.) Berk. & Broome (1846)

The neglected false truffle species Hydnotrya bailii Soehner (Ascomycetes, Discinaceae) is re-described and separated from its sister taxon Hydnotrya tulasnei by morphological and phylogenetic analyses based on internal transcribed spacer rDNA sequences. The most distinct morphological and ecological characters are small globose, rather than kidney-like, ascomata as known from the sister taxon H. tulasnei, strictly monoseriate ascospores and montane habitats. Phylogenetic analyses resulted in two clearly separated clusters that revealed the ectomycorrhizal specificity of H. bailii to Picea abies and that H. tulasnei is preferably associated to Fagus sylvatica. We also show that H. bailii was already present in mycorrhizal samples but until now could not be correctly assigned. Our analyses also indicate cryptic diversity within H. cerebriformis and other, morphologically not yet characterized, Hydnotrya groups. An emended determination key for all Hydnotrya species known from Central Europe is provided.

38: Setting the Mark: Establishing a Standard for Laparoscopic Visual Spatial Skills in Obstetrics and Gynecology Residents

Development of competence based advancement for surgical training is a desired alternative to the traditional “time-served” approach. A prior study demonstrated that a puff ball transfer drill or OSATS (objective structured assessment of technical skills) was a reliable and valid means of assessing laparoscopic visual-spatial skills. The goal of this study was to determine a benchmark for the puff ball transfer drill.

Isolation and mycelial growth of Diehliomyces microsporus: effect of culture medium and incubation temperature

The false truffle is one of the main problems in the production of the Agaricus brasiliensis in Brazil and the control of this fungal competitor has been rather difficult due to difficulties in the isolation and cultivation of this pathogen. This experiment was conducted in three stages, the first consisting of the isolation of Diehliomyces microsporus starting from portions of the fruiting body and through the ascospores suspension; second, D. microsporus cultivated in vitro at 15, 20, 25, 30 and 35ºC in six different culture media (CSDA, OCDA, PCDA, ODA, PDA, CDA); third, D. microsporus was inoculated on sterilized compost for formation of the fruiting body. The colony formation from tissue of D. microsporus starting from portions of fruiting body was more efficient than germination of the ascospores. Compost medium (CDA) allowed a larger diameter of the D. microsporus colony, followed by the medium made up of compost and potato mixture, favoring a denser composition. The largest mycelial growth speed of D. microsporus occurred when the culture was incubated at 28 and 30ºC. Incubation temperatures lower than 15ºC or above 35ºC inhibited the mycelial growth of D. microsporus completely. The fruiting bodies were obtained easily in sterilized compost and later inoculated along with mycelial competitor.

Use of a Robin's Nest as a Cache Site for Truffles by a Red Squirrel

A Turdus migratorius (American Robin) nest was found in southern New Brunswick, Canada, containing 52 mature sporocarps (“truffles”) of Elaphomyces granulatus (false truffle), a species of hypogeous fungus common across North America. Teeth marks on the truffles indicated they had been cached in the nest by a Tamiasciurus hudsonicus (red squirrel). The truffles appeared to have been air-dried before caching and were well preserved. Mean (± SD) weight of each truffle was 3.3 ± 1.4 g, with a total weight of cached material of 173 g. Although caching of epigeous fungus by squirrels is well documented in the literature, records of cached hypogeous fungi are relatively uncommon, and caches involving disused bird nests appear to be rarely encountered.

Geographic structure in a widespread plant–mycorrhizal interaction: pines and false truffles

Mutualistic interactions are likely to exhibit a strong geographic mosaic in their coevolutionary dynamics, but the structure of geographic variation in these interactions is much more poorly characterized than in host-parasite interactions. We used a cross-inoculation experiment to characterize the scales and patterns at which geographic structure has evolved in an interaction between three pine species and one ectomycorrhizal fungus species along the west coast of North America. We found substantial and contrasting patterns of geographic interaction structure for the plants and fungi. The fungi exhibited a clinal pattern of local adaptation to their host plants across the geographic range of three coastal pines. In contrast, plant growth parameters were unaffected by fungal variation, but varied among plant populations and species. Both plant and fungal performance measures varied strongly with latitude. This set of results indicates that in such widespread species interactions, interacting species may evolve asymmetrically in a geographic mosaic because of differing evolutionary responses to clinally varying biotic and abiotic factors.

Streptobacillus moniliformisEndocarditis

To the Editor: Streptobacillus moniliformis is a facultatively anaerobic, pleomorphic, gram-variable bacillus often seen in chains and as long unbranched filaments. It is found in the nasopharynx and oropharynx of wild and laboratory rats. Human infections result either from rodent bites (rat bite fever) or contaminated milk or other foods (Haverhill fever). The most common manifestations of infection are arthralgia, fever, and rash; endocarditis occurs as a rare complication (1). We report a case of S. moniliformis endocarditis in India in a patient with congenital heart disease. An 18-year-old man was admitted to the Department of Cardiology at the Government General Hospital in Chennai, India, in November 2005, with a fever of 2 months' duration with cough, epistaxis, palpitations, and persistent joint pain. His medical history indicated congenital heart disease with a ventricular septal defect. On physical examination, his blood pressure was 100/70 mm Hg, pulse rate was 100 beats/min, and temperature was 38.5°C. Laboratory tests showed a leukocyte count of 7,600/μL, a platelet count of 127,000/μL, and an erythrocyte sedimentation rate of 70 mm/h. An electrocardiogram showed normal sinus rhythm. A transthoracic echocardiogram demonstrated a ventricular septal defect and vegetations on the septal leaflet of the tricuspid valve. Three blood cultures were prepared, and treatment with antimicrobial drugs (intravenous penicillin G, 3 × 106 U every 6 h, and gentamicin, 50 mg every 8 h for 4 weeks) was initiated. The blood cultures were incubated at 37°C in an atmosphere of 5%–10% CO2. Characteristic white, downy, crumblike granules were observed on the surface of the erythrocytes in all 3 cultures within 18–24 h of incubation. Characteristic puff balls were seen after 48 h of incubation. Gram-stained smears showed gram-negative bacilli in long chains. Cultures were subcultured onto 5% sheep blood agar plates and MacConkey agar plates. The plates were incubated at 37°C in an atmosphere of 5%–10% CO2. After 18–24 h of incubation, growth was seen on the sheep blood agar plates. Colonies were 1–2 mm in diameter, gray, smooth, and butyrous. A Gram stain of these colonies identified gram-variable, pleomorphic coccobacilli that were negative for catalase, oxidase, urease, and citrate, and did not produce indole or reduce nitrate. Antimicrobial susceptibility testing was performed by using the Kirby-Bauer disk diffusion method according to recommendations of the National Committee for Clinical Laboratory Standards (2). The isolate was sensitive to penicillin G, ceftriaxone, cephalexin, amoxicillin, gentamicin, and erythromycin. The patient responded well to treatment and became afebrile within 48 h after initiation of therapy. Treatment with antimicrobial drugs was continued for 4 weeks. The blood cultures were negative when repeated after 2 weeks. The patient had an uneventful recovery and was discharged from the hospital. Rat bite fever is a zoonosis caused by either Streptobacillus moniliformis or Spirillum minus (1,3). S. moniliformis is found in the nasopharynx of small rodents, especially rats. Rats that are carriers have no symptoms but can effectively transmit the infection by bite or through infected body fluids such as urine. This patient had a history of living in a rat-infested area, and admitted having been bitten by a rat several months before the onset of symptoms. However, we considered it unlikely that disease contracted by a rat bite would take months to be manifested. Thus, it is more likely that he contracted the infection from food or water contaminated with rat excreta. Endocarditis is a rare complication of S. moniliformis infection, and cardiac valvular abnormalities have been reported in ≈50% of cases (4). This patient, however, had only a small ventricular septal defect. This is the first report of S. moniliformis endocarditis from India.

Notes on Scleroderma , Puff balls and Geastrum of Azad Jammu and Kashmir, Pakistan

Six species of mushrooms allied to the Family Sclerodermataceae, Lycoperdaceae and Geastraceae have been described for the first time from Azad Jammu and Kashmir. These are Scleroderma aurantium, Calvatia verrucosia sp. nov., Lycoperdon pedicellaton sp. nov. L. sphaericon sp. nov., L. echinulaton sp. nov., and Geastrum heptaplex sp. nov.

Calcaelin, a new protein with translation-inhibiting, antiproliferative and antimitogenic activities from the mosaic puffball mushroom Calvatia caelata.

Fresh fruiting bodies of the mushroom Calvatia caelata, commonly known as the mosaic puffball, were extracted with 10 mM Tris-HCl buffer (pH 7.2). The extract was subjected to a chromatographic procedure which has been successfully used to isolate ribosome-inactivating proteins from plants. It was first applied to an Affi-gel blue gel column previously equilibrated and eluted with 10 mM Tris-HCl buffer. The adsorbed fraction eluted with 1.5 M NaCl in the buffer was then purified by chromatography on DEAE-cellulose in the same buffer. The unadsorbed fraction yielded essentially a single peak when it was chromatographed on a column of Mono S. When the peak was then gel filtered on Superdex 75, it yielded a homogeneous peak with a molecular mass of 39 kDa. The protein, which was designated calcaelin, was dissociated into two subunits with a molecular mass of 19 kDa and 20 kDa, respectively, in SDS-polyacrylamide gel electrophoresis. The two bands exhibited the same N-terminal amino acid sequence which was somewhat similar to those of plant ribosome-inactivating proteins. A lesser degree of sequence homology to the fungal ribosome inactivating proteins alpha-sarcin and restrictocin was detected. Calcaelin inhibited translation in rabbit reticulocyte lysate with an IC 50 value of 4 nM and displayed a heat-labile RNase activity of 1.58 U/mg toward yeast tRNA. It exhibited an antimitogenic activity toward mouse splenocytes, and it reduced the viability of breast cancer cells. There was no hemagglutinating, antibacterial or antifungal activity.

Flutolanil for Control of Necrotic Fairy Rings on Bermudagrass Putting Greens

Type 1 (necrotic) fairy rings in turfgrass result in dead or badly damaged grass. This type of fairy ring is a severe problem on golf course greens as they interfere with the aesthetics and playability of the putting surface. In Florida, Lycoperdon spp., basidiomycetes that produce puffball mushrooms, have been implicated as a common cause of Type 1 fairy rings on hybrid bermudagrass (Cynodon dactylon × C. transvaalensis) putting greens. The fungicide flutolanil has basidiomycetes as the sole fungal target. It is also the only carboxin-related fungicide registered for use on turfgrass. Two experiments were conducted to examine the effect of flutolanil as a curative and preventive treatment for fairy ring caused by Lycoperdon. One experiment, established after the rings were present, determined that flutolanil significantly reduced mushroom production. The second experiment was conducted on a golf course that had experienced Type 1 fairy rings previously. One-half of each of nine putting greens was treated with flutolanil on a preventive basis. The other half of each green served as an untreated control. Type 1 fairy rings, due to Lycoperdon, developed only on the untreated control half of each green. These experiments confirm that flutolanil does have curative and preventive activity against Lycoperdon spp. that cause Type 1 fairy rings.

Enrichment and isolation of Beggiatoa spp. From rice eccosystem

Beggiatoa, an organism capable of oxidizing hydrogen sulfide has been isolated from rhizosphere of rice grown under submerged ecosystem. The isolation procedure involves prior enrichment with extracted hay and soil extract under various treatments viz., addition of acetate or sodium sulfide or acetate + sodium sulfide, diluted and undiluted soil extract. The results indicated that diluted (1:1) soil extract with sodium sulfide enrichment had registered higher number of 22.5 trichomes ml', followed by acetate + sodium sulfide treatment, apart from reducing the bacterial and protozoan contaminants to a minimal level. Formation of puff balls and mat in the sub-surface of the enrichment medium suggest that they are microaerophilic and grown in the interface between oxygen and hydrogen sulfide environments. Growth of Beggiatoa was also observed in Beggiatoa medium 2 as circuitance type of colony. Morphological characterizations of this isolate showed its close resemblances with that of B. alba

Antiproliferative and Antimitogenic Activities in a Peptide from Puffball Mushroom Calvatia caelata

A peptide with a molecular weight of 8 kDa and an N-terminal sequence closely resembling that of ubiquitin was isolated from fruiting bodies of the mosaic puffball mushroom Calvatia caelata. The peptide was purified using a protocol that involved ion-exchange chromatography on DEAE–cellulose, affinity chromatography on Affi-gel blue gel, and ion-exchange chromatography on Mono S. The peptide inhibited translation in the cell-free rabbit reticulocyte lysate system and exhibited N-glycosidase activity. It potently inhibited proliferation of spleen cells with an IC50 of about 100 nM as indicated by the suppression of [methyl-3H]thymidine uptake. The viability of breast cancer cells was reduced to half at a ubiquitin concentration of about 100 nM.