Roquefortine C Occurrence in Blue Cheese

Total Page:16

File Type:pdf, Size:1020Kb

Roquefortine C Occurrence in Blue Cheese 246 Journalof FoodProtection, Vol. 64, No. 2, 2001, Pages 246– 251 Copyright q,International Association forFood Protection RoquefortineC Occurrence inBlue Cheese CARLO FINOLI, 1 ANGELA VECCHIO, 2* ANTONIETTA GALLI, 2 AND IVAN DRAGONI 3 1Dipartimentodi Ingegneria e TecnologieAgro-Forestali, Universita `degliStudi di Palermo, viale delle Scienze 10,90128 Palermo, Italy; 2Dipartimentodi Scienze eTecnologieAlimentari e Microbiologiche,Universita `degliStudi di Milano, via Celoria 2, 20133 Milano, Italy; and 3Istitutodi Ispezione degli Alimenti di Origine Animale, Universita `degliStudi di Milano, via Celoria 6, 20133 Milano, Italy MS00-186: Received 9June2000/ Accepted 6September2000 ABSTRACT Downloaded from http://meridian.allenpress.com/jfp/article-pdf/64/2/246/1671860/0362-028x-64_2_246.pdf by guest on 27 September 2021 Severalstrains of Penicillium areused for the production of mold-ripenedcheeses, and some of themare able to produce mycotoxins.The aims of the research were the determination of roquefortine C andPR toxinin domestic and imported blue cheeses,the identi cation of thepenicillia used as starter,and the investigation of their capacity for producing toxins in culture media.Roquefortine C wasalways found in the cheeses at levels ranging from 0.05 to 1.47 mg/ kg,whereas the PR toxinwas neverfound. The identi cation of thefungal strains present in thedomestic cheeses included Penicilliumglabrum, Penicillium roqueforti, and Penicilliumcyclopium inthe Gorgonzola ‘ ‘dolce’’ and Penicilliumroqueforti inthe Gorgonzola ‘ ‘naturale’’; inone case, the presence of Penicilliumcrustosum wasobserved. The strains isolated from the foreign cheeses belonged to P.roqueforti. Thestrains were able to producebetween 0.18 and 8.44 mg/ literof roquefortinein yeastextract sucrose medium andbetween 0.06 and 3.08 mg/ literand less than 0.05 mg/ literwhen inoculated in milk at 20 8Cfor14 days and 4 8C for 24 days,respectively. Linear relations between production of roquefortinein culture media and cheeses did not emerge. PR toxin rangedfrom less than 0.05 to 60.30 mg/ literin yeast extract sucrose medium and was produced in milk at 20 8C from only onestrain. The low levels and the relatively low toxicity of roquefortine make the consumption of blue cheese safe for the consumer. Mostpenicillia reside in soil or onvegetables, and their aspatulin, penicillic acid, mycophenolic acid, isofumigo- presencein foods usually is connected with an accidental clavinA andB, roquefortine C, and PR toxin (11). The contaminationduring production; in some cases, such as presentresearch has been limited to the last two toxins be- fermentedsalami and ripened cheese, they are addedto causethey are the mycotoxins more easily produced from developspeci c organolepticcharacteristics (7). Some thecultures used as starters for theproduction of Gorgon- cheesesare manufactured with strains of Penicilliumro- zola.Roquefortine has neurotoxic properties (34); PR toxin queforti, i.e.,the blue-veined cheeses such as Gorgonzola, inhibitsRNA andprotein synthesis (18, 19) andis carci- Roquefort,Bleu de Bresse, Cabrales, Danablu, and Edel- nogenicfor rats (22) andmutagenic for Salmonella Typhi- pilzka¨se,and strains of Penicilliumcamemberti are used for murium (33). whitesurface mold cheeses such as Camembert, Brie, and Fungalstrains used as starters in the dairy industry Coulommiers.Once these molds were naturallyfound in were ableto produce roquefortine in vitro (10, 26). The themilk and cheese, whereas today spore suspensions are presenceof roquefortinewas foundalso in cheeses (15, 26, directlysprayed on the surface of cheeses or added to the 28, 32, 35).PRtoxin was isolated rst inthe extracts of milkor curd. The most important biochemical characteris- culturemedium of P.roqueforti by Wei et al. (36) in 1973. ticsare the proteolytic activity, which plays a fundamental Itwas neverfound in blue cheeses (9,23, 26, 27) , because roleduring blue cheese ripening, and the lipolytic activity, itreacts with nitrogenous compounds, converting itself to whichinduces triglyceride hydrolysis and subsequent b- PR-imine,which has a lowertoxicity (1).Siemensand Za- ketoacidand methylketone formation. wistowski (30),ina studyon derivatives of the PR toxin P.camemberti and P.roqueforti were usedwithout incheeses, reported PR-imine at levels of 19 to 42 mg/kg particularproblems in cheesemaking until Gibel et al. (14) in50 ofthe60 samplesanalyzed. The authors demonstrated publisheda studyin whichthey assumed that such cultures thatin vitro, in the presence of bovine serum, the conver- couldrepresent a riskfor consumerhealth because of the sionof thisderivative in themore dangerous original parent formationof toxic metabolites. The authors suggested that was possible.PR toxinwas foundin culture media up to P.camemberti extractsseemed to be able to induce carci- 200mg/ liter (9). nogeniceffects in mice. P.roqueforti, atpresentreclassi ed Gorgonzolabelongs to the group of the blue cheeses afterpolymerase chain reaction identi cation into three spe- havingin common the speci c characteristicof the for- cies named P.roqueforti,Penicillium carneum, and Peni- mationof bluegreenish veins in their interior ,producedby cilliumpaneum (5) ,isable to produce several toxins, such mold of the P.roqueforti type,with tonality of moreor less intensecolor (12,13, 24, 25) .It isa cheeseof ‘ ‘denomin- *Authorfor correspondence. Tel: 0039022367181; Fax: 0039022361576; azionedi origine’’ (6),originatingfrom Gorgonzola,a town E-mail: [email protected]. nearMilan, and its production is governed by appropriate J.FoodProt., Vol. 64, No. 2 ROQUEFORTINE COCCURRENCE INBLUECHEESE 247 regulationsestablished by theConsortium for theProtection TABLE 1. Mycotoxinrecovery percentages and standard devia- ofGorgonzola Cheese. Only the dairies authorized by the tions consortiumcan market Gorgonzola cheese with the brand Added name ‘‘g,’’whichguarantees its authenticity. Depending on amount Cheese YES medium Milk themanufacturing processes, two varieties of Gorgonzola Mycotoxin (mg/kg) (%) (%) (%) canbe obtained:a traditionaltype, ‘ ‘Gorgonzolanaturale,’ ’ Roquefortine25 97 6 5.2 61 6 5.7 53 6 4.2 witha consistingand homogenous texture of the paste, 200 101 6 4.3 65 6 5.1 62 6 4.8 sometimesslightly chalky, a strongtaste, with molds of a shininggreen color ,verydeveloped and uniformly distrib- PR toxin 500 45 6 4.7 65 6 2.3 48 6 3.7 utedin the paste; and a creamytype, ‘ ‘Gorgonzoladolce,’ ’ 1000 46 6 3.4 69 6 1.8 50 6 2.6 witha soft,smeared texture of the paste, sometimes with smalleyelike holes, a sweettaste, with green-colored molds littledeveloped and uniformly distributed in thepaste. The matography(TLC) andin high-performance liquid chromatogra- 7 Downloaded from http://meridian.allenpress.com/jfp/article-pdf/64/2/246/1671860/0362-028x-64_2_246.pdf by guest on 27 September 2021 productionof Gorgonzolacheese is ofabout4.2 3 10 kg/ phy(HPLC) eluentfor roquefortine (3) andin methanol for PR yearand 28% is exported (data from theConsortium for toxinfor HPLC analysis.Working standard solutions of roque- theProtection of Gorgonzola Cheese). fortineand PR toxinwere prepared from crystalline toxins (Sigma Theobjectives of this study were toinvestigate the ChemicalCo., St. Louis, Mo.). The purity of the standards was ability of Penicillium speciesto produce mycotoxins in blue determinedin absolute ethanol by UV analysisusing a spectro- cheeses,to isolateand identify the molds, and to determine photometerUV/ VIS Lambda15 Perkin Elmer (Perkin Elmer ifthesemolds produce mycotoxins in variousculture media Corp.,Norwalk, Conn.) and compared with reported spectra (8). incubatedin different conditions. BeforeHPLC analysis,all samples were analyzed by TLC. MATERIALS AND METHODS TLC analysis. HPTLC Kieselgel60 F 254 andFertigplatten Kieselgel60 (Merck) (10 by 10 cm), activated at 110 8C for 2 h, TwentyItalian Gorgonzola cheeses, supplied by the Consor- wereused for TLC ofroquefortine and PR toxin,respectively. tiumfor the Protection of Gorgonzola cheese with technical in- Thedevelopment was monodimensional using, in sequence, n- formation,and 10 foreign blue cheeses, purchased in various Eu- hexane,diethyl ether, and chloroform– methanol– ammonium hy- ropeancountries, have been analyzed. droxide(9:1:0.1) for roquefortine and n-hexane, diethyl ether, and toluene–ethyl acetate– acetic acid (6:4:1) for PR toxin.After de- Isolationand classication of penicillia. Themolds were velopment,plates were air dried. The roquefortine plates were isolatedafter extracting a portionof thegreen zone of the cheese observedunder UV light(UVLS-58 mineral light multiband 254 witha sterileneedle. The mycelium was transferred to a maltagar to366 nm, San Gabriel, Calif.) at 254 nm, and the toxin appeared plateand incubated at 20 8Cfor7 daysaccording to Pitt and Hock- likea darkspot. After spraying plates with 1% CeSO 4 in 6 N ing (21).Thepurity of the strain isolated was tested with a mi- sulfuricacid, roquefortine formed an orange spot (26). For the croscope,and the strain was kept on malt agar slant at 4 8C. The detectionof PR toxin,plates after exposure to UV lightat 254 identication and classi cation of molds were carried out using nmfor about 1 minwere observed at 366 nm; the dark blue thefollowing media: malt extract agar ,25%glycerol nitrate agar , uorescentspot changed to green uorescence (4). andCzapek yeast extract agar according to Pitt and Hocking (21). HPLC analysis. Roquefortineanalysis was carried out ac- Mycotoxinproduction. Thecapability of
Recommended publications
  • Incorporating Fermentation Into Undergraduate Laboratory Courses
    Athens Journal of Sciences- Volume 2, Issue 4 – Pages 257-264 Incorporating Fermentation into Undergraduate Laboratory Courses By Claire Gober Madeleine Joullie† Laboratory courses in universities have a responsibility to introduce current research practices and trends in scientific research to adequately prepare students for work in the field. One such research practice gaining popularity in recent years is that of green chemistry. Since the 1960s, increasing concern over the release of toxic chemicals into the environment has led to a push for more environmentally responsible chemistry. A growing faction of chemists has begun to adopt methods to eliminate chemical waste and support green chemistry. Fermentation is an ideal technique to demonstrate environmentally sustainable chemistry in an undergraduate laboratory class. Fermentation of complex natural products, as opposed to traditional organic synthesis, is beneficial as it supports a number of principles of green chemistry; it is conducted at ambient temperature and pressure, uses inexpensive and innocuous materials, makes use of renewable resources, and does not require a fume hood. Skills implemented during fermentation can be easily taught to upper-level Chemistry and Biochemistry undergraduate students, who typically have limited exposure to complex natural products in their coursework. Such a course would be interdisciplinary in nature, incorporating fungal biology and metabolism as well as organic chemistry. Students would learn a variety of skills, including growth media selection and preparation, inoculation of fungal cultures, extraction of natural products, and purification and characterization of metabolites. Experiments of this nature would allow for discussions of several areas of research: green chemistry, natural products and their application to medicine, identification of functional groups in complex molecules by spectroscopy, and introduction to biochemistry and metabolism.
    [Show full text]
  • Penitrem a Peer-Reviewed Journal Articles
    ________________________________________________________________________ ________________________________________________________________________ Penitrem A Peer-Reviewed Journal Articles Literature Review by Lisa Petrison, Ph.D. Neurotoxic Effects: Berntsen H. F., Wigestrand M. B., Bogen I. L., Fonnum F., Walaas S. I., Moldes-Anaya A.. Mechanisms of penitrem-induced cerebellar granule neuron death in vitro: possible involvement of GABAA receptors and oxidative processes. Neurotoxicology. 2013;35:129–136. Moldes-Anaya Angel, Rundberget Thomas, Fæste Christiane K., Eriksen Gunnar S., Bernhoft Aksel. Neurotoxicity of Penicillium crustosum secondary metabolites: tremorgenic activity of orally administered penitrem A and thomitrem A and E in mice. Toxicon. 2012;60:1428–1435. Moldes-Anaya Angel S., Fonnum Frode, Eriksen Gunnar S., Rundberget Thomas, Walaas S. Ivar, Wigestrand Mattis B.. In vitro neuropharmacological evaluation of penitrem-induced tremorgenic syndromes: importance of the GABAergic system. Neurochemistry international. 2011;59:1074–1081. Lu Hai-Xia X., Levis Hannah, Liu Yong, Parker Terry. Organotypic slices culture model for cerebellar ataxia: potential use to study Purkinje cell induction from neural stem cells. Brain research bulletin. 2011;84:169–173. Namiranian Khodadad, Lloyd Eric E., Crossland Randy F., et al. Cerebrovascular responses in mice deficient in the potassium channel, TREK-1. American journal of physiology. Regulatory, integrative and comparative physiology. 2010;299. Ohno Akitoshi, Ohya Susumu, Yamamura Hisao,
    [Show full text]
  • Roquefortine C (SML0406)
    Roquefortine C from Penicillium roqueforti Catalog Number SML0406 Storage Temperature –20 °C CAS RN 58735-64-1 Precautions and Disclaimer Synonym: Roquefortine This product is for R&D use only, not for drug, household, or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices. Preparation Instructions Roquefortine C is soluble in methanol (10 mg/mL), DMSO (10 mg/mL), ethyl acetate (1 mg/mL), and chloroform (1 mg/mL). Storage/Stability Store the product sealed at –20 °C. Under these conditions the product is stable for at least 4 years. A DMSO solution (10 mg/mL) is stable for 3 months at Product Description –20 °C. Molecular formula: C22H23N5O2 Molecular weight: 389.45 References 1. Kopp-Holtwiesche, B., and Rehm, H.J., Roquefortine C is a paralytic neurotoxin with a Antimicrobial action of roquefortine. J. Environ. dioxopiperazine structure produced by a diverse range Pathol. Toxicol. Oncol., 10, 41-44 (1990). 1 of fungi, most notably Penicillium species. It has been 2. Wagener, R.E. et al., Penitrem A and Roquefortine 2 found in blue cheese and in many other food products Production by Penicillium commune. App. Environ. 3 due to natural occurrence and contamination. Microbiol., 39, 882-887 (1980). 3. Shangguan, N. et al., The total synthesis of Roquefortine C was found to be active on a wide range roquefortine C and a rationale for the of organisms. It inhibits the growth of Gram-positive thermodynamic stability of isoroquefortine C over bacteria, and cockerels treated with roquefortine lost roquefortine C. J. Am.
    [Show full text]
  • Occurrence and Properties of Thiosilvatins
    marine drugs Review Occurrence and Properties of Thiosilvatins Maria Michela Salvatore 1 , Rosario Nicoletti 2,3 , Marina DellaGreca 1 and Anna Andolfi 1,* 1 Department of Chemical Sciences, University of Naples ‘Federico II’, 80126 Naples, Italy; [email protected] (M.M.S.); [email protected] (M.D.) 2 Council for Agricultural Research and Economics, Research Centre for Olive, Citrus and Tree Fruit, 81100 Caserta, Italy; [email protected] 3 Department of Agriculture, University of Naples ‘Federico II’, 80055 Portici, Italy * Correspondence: andolfi@unina.it; Tel.: +39-081-2539179 Received: 27 September 2019; Accepted: 21 November 2019; Published: 26 November 2019 Abstract: The spread of studies on biodiversity in different environmental contexts is particularly fruitful for natural product discovery, with the finding of novel secondary metabolites and structural models, which are sometimes specific to certain organisms. Within the large class of the epipolythiodioxopiperazines, which are typical of fungi, thiosilvatins represent a homogeneous family that, so far, has been reported in low frequency in both marine and terrestrial contexts. However, recent observations indicate that these compounds have been possibly neglected in the metabolomic characterization of fungi, particularly from marine sources. Aspects concerning occurrence, bioactivities, structural, and biosynthetic properties of thiosilvatins are reviewed in this paper. Keywords: thiosilvatins; secondary metabolites; diketopiperazines; epipolythiodioxopiperazines; fungi; biological activities 1. Introduction A huge chapter on research on biodiversity is represented by studies concerning the biochemical properties of the manifold organisms which are part of natural ecosystems. Novel secondary metabolites are continuously discovered, disclosing a surprising chemodiversity in terms of both structural and biosynthetic aspects.
    [Show full text]
  • By Penicillium Paneum Frisvad and Penicillium Roqueforti Thom Isolated from Baled Grass Silage in Ireland
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Online Research Database In Technology 9268 J. Agric. Food Chem. 2006, 54, 9268−9276 Mycotoxins and Other Secondary Metabolites Produced in Vitro by Penicillium paneum Frisvad and Penicillium roqueforti Thom Isolated from Baled Grass Silage in Ireland MARTIN O’BRIEN,*,†,# KRISTIAN F. NIELSEN,‡ PADRAIG O’KIELY,† PATRICK D. FORRISTAL,§ HUBERT T. FULLER,# AND JENS C. FRISVAD‡ Teagasc, Grange Beef Research Centre, Dunsany, Co. Meath, Ireland; Center for Microbial Biotechnology, BioCentrum-DTU, Technical University of Denmark, Building 221, DK-2800 Kgs. Lyngby, Denmark; Teagasc, Crops Research Centre, Oak Park, Co. Carlow, Ireland; and School of Biology and Environmental Science, College of Life Sciences, University College Dublin, Belfield, Dublin 4, Ireland Secondary metabolites produced by Penicillium paneum and Penicillium roqueforti from baled grass silage were analyzed. A total of 157 isolates were investigated, comprising 78 P. paneum and 79 P. roqueforti isolates randomly selected from more than 900 colonies cultured from bales. The findings mostly agreed with the literature, although some metabolites were not consistently produced by either fungus. Roquefortine C, marcfortine A, and andrastin A were consistently produced, whereas PR toxin and patulin were not. Five silage samples were screened for fungal metabolites, with two visually moldy samples containing up to 20 mg/kg of roquefortine C, mycophenolic acid, and andrastin A along with minor quantities (0.1-5 mg/kg) of roquefortines A, B, and D, festuclavine, marcfortine A, and agroclavine. Three visually nonmoldy samples contained low amounts of mycophenolic acid and andrastin A.
    [Show full text]
  • Putative Neuromycotoxicoses in an Adult Male Following Ingestion of Moldy Walnuts
    Short research communication Putative neuromycotoxicoses in an adult male following ingestion of moldy walnuts Authors: C. J. Botha1 C. M. Visagie2 M. Sulyok3 Affiliations: 1 Department of Paraclinical Sciences, Faculty of Veterinary Science, University of Pretoria, Private bag X04, Onderstepoort, 0110, South Africa 2 Biosystematics Division, Agricultural Research Council – Plant Health and Protection, Private Bag X134, Queenswood, Pretoria, 0121, South Africa 3 Centre for Analytical Chemistry, Department of Agrobiotechnology (IFA-Tulln), University of Natural Resources and Life Sciences, Konrad Lorenzstr. 20, A-3430, Tulln, Austria CJ Botha [email protected] Tel.: +27 125298023; fax +27 125298304. ORCID CJ Botha 0000 0003 1535 9270 ORCID CM Visagie 0000-0003-2367-5353 ORCID M Sulyok 0000-0002-3302-0732 1 Abstract A tremorgenic syndrome occurs in dogs following ingestion of moldy walnuts and Penicillium crustosum has been implicated as the offending fungus. This is the first report of suspected moldy walnut toxicosis in man. An adult male ingested approximately 8 fungal-infected walnut kernels and after 12 hours experienced tremors, generalized pain, incoordination, confusion, anxiety and diaphoresis. Following symptomatic and supportive treatment at a local hospital the man made an uneventful recovery. A batch of walnuts (approximately 20) were submitted for mycological culturing and identification as well as for mycotoxin analysis. Penicillium crustosum Thom was the most abundant fungus present on walnut samples, often occurring as monocultures on isolation plates. Identifications were confirmed with DNA sequences. The kernels and shells of the moldy walnuts as well as P. crustosum isolates plated on Yeast Extract Sucrose (YES) and Czapek Yeast Autolysate (CYA) agars and incubated in the dark at 25 ºC for 7 days were screened for tremorgenic mycotoxins and known P.
    [Show full text]
  • Preservation Stress Resistance of Melanin Deficient Conidia From
    Seekles et al. Fungal Biol Biotechnol (2021) 8:4 https://doi.org/10.1186/s40694-021-00111-w Fungal Biology and Biotechnology RESEARCH Open Access Preservation stress resistance of melanin defcient conidia from Paecilomyces variotii and Penicillium roqueforti mutants generated via CRISPR/Cas9 genome editing Sjoerd J. Seekles1,2, Pepijn P. P. Teunisse1,2, Maarten Punt1,3, Tom van den Brule1,4, Jan Dijksterhuis1,4, Jos Houbraken1,4, Han A. B. Wösten1,3 and Arthur F. J. Ram1,2* Abstract Background: The flamentous fungi Paecilomyces variotii and Penicillium roqueforti are prevalent food spoilers and are of interest as potential future cell factories. A functional CRISPR/Cas9 genome editing system would be benefcial for biotechnological advances as well as future (genetic) research in P. variotii and P. roqueforti. Results: Here we describe the successful implementation of an efcient AMA1-based CRISPR/Cas9 genome edit- ing system developed for Aspergillus niger in P. variotii and P. roqueforti in order to create melanin defcient strains. Additionally, kusA− mutant strains with a disrupted non-homologous end-joining repair mechanism were created to further optimize and facilitate efcient genome editing in these species. The efect of melanin on the resistance of conidia against the food preservation stressors heat and UV-C radiation was assessed by comparing wild-type and melanin defcient mutant conidia. Conclusions: Our fndings show the successful use of CRISPR/Cas9 genome editing and its high efciency in P. vari- otii and P. roqueforti in both wild-type strains as well as kusA− mutant background strains. Additionally, we observed that melanin defcient conidia of three food spoiling fungi were not altered in their heat resistance.
    [Show full text]
  • The Brla Gene Deletion Reveals That Patulin Biosynthesis Is Not Related to Conidiation in Penicillium Expansum
    International Journal of Molecular Sciences Article The brlA Gene Deletion Reveals That Patulin Biosynthesis Is Not Related to Conidiation in Penicillium expansum Chrystian Zetina-Serrano, Ophélie Rocher, Claire Naylies, Yannick Lippi , Isabelle P. Oswald , Sophie Lorber and Olivier Puel * Toxalim (Research Centre in Food Toxicology), Université de Toulouse, INRAE, ENVT, INP-Purpan, UPS, 31027 Toulouse, France; [email protected] (C.Z.-S.); [email protected] (O.R.); [email protected] (C.N.); [email protected] (Y.L.); [email protected] (I.P.O.); [email protected] (S.L.) * Correspondence: [email protected]; Tel.: +33-582-066-336 Received: 7 August 2020; Accepted: 8 September 2020; Published: 11 September 2020 Abstract: Dissemination and survival of ascomycetes is through asexual spores. The brlA gene encodes a C2H2-type zinc-finger transcription factor, which is essential for asexual development. Penicillium expansum causes blue mold disease and is the main source of patulin, a mycotoxin that contaminates apple-based food. A P. expansum PeDbrlA deficient strain was generated by homologous recombination. In vivo, suppression of brlA completely blocked the development of conidiophores that takes place after the formation of coremia/synnemata, a required step for the perforation of the apple epicarp. Metabolome analysis displayed that patulin production was enhanced by brlA suppression, explaining a higher in vivo aggressiveness compared to the wild type (WT) strain. No patulin was detected in the synnemata, suggesting that patulin biosynthesis stopped when the fungus exited the apple. In vitro transcriptome analysis of PeDbrlA unveiled an up-regulated biosynthetic gene cluster (PEXP_073960-PEXP_074060) that shares high similarity with the chaetoglobosin gene cluster of Chaetomium globosum.
    [Show full text]
  • Synthetic Studies of Roquefortine C: Synthesis of Isoroquefortine
    Synthetic studies of roquefortine C: SPECIAL FEATURE Synthesis of isoroquefortine C and a heterocycle David J. Richard, Bruno Schiavi, and Madeleine M. Joullie´ * Department of Chemistry, University of Pennsylvania, 231 South 34th Street, Philadelphia, PA 19104-6323 Edited by Kyriacos C. Nicolaou, The Scripps Research Institute, La Jolla, CA, and approved April 14, 2004 (received for review April 9, 2004) The syntheses of isoroquefortine C and a related heterocycle were achieved by implementation of both intra- and intermolecular vinyl amidation reactions. These accomplishments represent a signifi- cant advance in the use of these strategies in the generation of complex molecules. etabolites produced by fungi represent one of the largest Mclasses of natural products. Such compounds vary widely in their structural composition and have found application in pharmaceuticals such as antibiotics, immunosuppressants, anti- fungal agents, and growth promoters (1). Other congeners Fig. 1. The roquefortine class of natural products. within this class display biological properties harmful to humans and other animals. These natural products have been classified as mycotoxins and are produced as secondary metabolites by an containing hemins but no potency toward Gram-negative or- CHEMISTRY array of soilborne and airborne fungi (2). Mycotoxins have been ganisms (15). Additional studies led to the finding that roque- isolated as contaminants of a variety of grain products and have fortine C inhibited bacterial RNA synthesis but only modestly been a topic of great interest for scientists concerned with affected DNA and protein production (16). veterinary health and agricultural safety. The lack of consistent toxicological data, along with the In most cases, eradication of the fungal infection responsible ubiquitous nature of P.
    [Show full text]
  • Patulin Fact Sheet
    Patulin Fact Sheet Background Information: Patulin takes it’s name from the fungus from which it was first isolated, Penicillium patulum. It is most likely to occur in moldy fruits, such as apples, but may also be found in grains, especially wet grains, and silages. Patulin may be associated with problems in silages, however the data on that aspect has not been well established. Since Patulin is associated with storage molds, it should be considered when investigating silage problems, specifically regarding dairy catte. Can co-contaminate feedstuffs with other Penicillium toxins, such as, Ochratoxin A (OTA), Citrinin, Roquefortine C, Mycophenolic Acid (MPA), and Cyclopiazonic Acid. Major crops affected: Corn, Barley, Wheat, & Rye, their associated silages, and Fruits. Associated Mold: Penicillium sp., Aspergillus sp., and Byssochlamys sp. Conditions favoring production: Penicillium is a major silage mold and may be a greater silage problem because it can grow at lower pH than do other molds. Considered to be more prevalent from a storage mold situation, rather than field/growing condition mold situation. Symptoms: Patulin is an antibiotic against gram-positive bacteria. In ruminants, Patulin has been shown to reduce VFA production, fiber digestion, and bacterial yield. Also, nephrotoxic (kidney) and immunotoxic effects. Gastrointestinal symptoms like gastric ulcers, intestinal hemorrages, lesions in the duodenum, and alteration of intestinal barrier function. Detection Limit: 200 ppb Dairyland Labs Packages that include Patulin: • Mycotoxin Select Package • Mycotoxin Complete Package Sources Diaz, D.E., W.M. Hagler, and L.W. Whitlow. “Mycotoxins in Feeds.” Feedstuffs. 15 Sep. 2010. Gallo, A., G. Giubuerti, J.C. Frisvad, T. Bertuzzi, and K.F.
    [Show full text]
  • Targeted and Untargeted Analysis of Secondary Fungal Metabolites by Liquid Chromatography-Mass Spectrometry
    Targeted and untargeted analysis of secondary fungal metabolites by liquid chromatography-mass spectrometry Svetlana V. Malysheva Promoter: Prof. Dr. Sarah De Saeger Co-promoters: Prof. Dr. Irina Yu. Goryacheva Dr. José Diana Di Mavungu 2013 THESIS SUBMITTED IN FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR IN PHARMACEUTICAL SCIENCES PROEFSCHRIFT VOORGELEGD TOT HET BEKOMEN VAN DE GRAAD VAN DOCTOR IN DE FARMACEUTISCHE WETENSCHAPPEN Targeted and untargeted analysis of secondary fungal metabolites by liquid chromatography-mass spectrometry Svetlana V. Malysheva Promoter: Prof. Dr. Sarah De Saeger Co-promoters: Prof. Dr. Irina Yu. Goryacheva Dr. José Diana Di Mavungu 2013 THESIS SUBMITTED IN FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR IN PHARMACEUTICAL SCIENCES PROEFSCHRIFT VOORGELEGD TOT HET BEKOMEN VAN DE GRAAD VAN DOCTOR IN DE FARMACEUTISCHE WETENSCHAPPEN PhD thesis Title: Targeted and untargeted analysis of secondary fungal metabolites by liquid chromatography – mass spectrometry Author: Svetlana V. Malysheva Year: 2013 Pages: 338 ISBN: 978-94-6197-128-9 Printer: University Press, Zelzate, Belgium Refer to this thesis as follows: Malysheva SV (2013). Targeted and untargeted analysis of secondary fungal metabolites by liquid chromatography – mass spectrometry. Thesis submitted in fulfilment of the requirements of the degree of Doctor (Ph.D.) in Pharmaceutical Sciences. Faculty of Pharmaceutical Sciences, Ghent University Promoter: Prof. Sarah De Saeger (Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium) Co-promoters: Prof. Irina Yu. Goryacheva (Chemistry Institute, Saratov State University, Russia) Dr. José Diana Di Mavungu (Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium) Members of the reading committee: Prof. Ann Van Schepdael (Faculty of Pharmaceutical Sciences, KU Leuven, Belgium) Prof.
    [Show full text]
  • Modelling Fungal Growth, Mycotoxin Production and Release in Grana Cheese
    microorganisms Article Modelling Fungal Growth, Mycotoxin Production and Release in Grana Cheese Marco Camardo Leggieri 1 , Amedeo Pietri 2 and Paola Battilani 1,* 1 Department of Sustinable Crop Production, Università Cattolica del Sacro Cuore, Via E. Parmense, 84, 29122 Piacenza, Italy; [email protected] 2 Department of Animal Science, Food and Nutrition, Università Cattolica del Sacro Cuore, Via E. Parmense, 84, 29122 Piacenza, Italy; [email protected] * Correspondence: [email protected]; Tel.: +39-0523-599-254 Received: 29 November 2019; Accepted: 31 December 2019; Published: 2 January 2020 Abstract: No information is available in the literature about the influence of temperature (T) on Penicillium and Aspergillus spp. growth and mycotoxin production on cheese rinds. The aim of this work was to: (i) study fungal ecology on cheese in terms of T requirements, focusing on the partitioning of mycotoxins between the rind and mycelium; and (ii) validate predictive models previously developed by in vitro trials. Grana cheese rind blocks were inoculated with A. versicolor, P. crustosum, P. nordicum, P. roqueforti, and P. verrucosum, incubated at different T regimes (10–30 ◦C, step 5 ◦C) and after 14 days the production of mycotoxins (ochratoxin A (OTA); sterigmatocystin (STC); roquefortine C (ROQ-C), mycophenolic acid (MPA), Pr toxin (PR-Tox), citrinin (CIT), cyclopiazonic acid (CPA)) was quantified. All the fungi grew optimally around 15–25 ◦C and produced the expected mycotoxins (except MPA, Pr-Tox, and CIT). The majority of the mycotoxins produced remained in the mycelium (~90%) in three out of five fungal species (P. crustosum, P. nordicum, and P.
    [Show full text]