scholarly journals Penicillium Species Isolated From Cocoa, Coffee Beans, and Dried Cassava in Yogyakarta Indonesia and Their Ochratoxin Production

2014 ◽  
Vol 13 (1) ◽  
pp. 1 ◽  
Author(s):  
Mona Nur Moulia ◽  
Sigit Setyabudi ◽  
Baharuddin Salleh ◽  
Endang S. Rahayu
Keyword(s):  
Ochratoxin A ◽  
Morphological Characteristics ◽  
Penicillium Citrinum ◽  
Molecular Characteristics ◽  
Cocoa Beans ◽  
Penicillium Species ◽  
Coffee Beans ◽  
Pcr Products ◽  
Yeast Extract Sucrose ◽  
Penicillium Paneum

The presence of Penicillium in cocoa and coffee beans, and dried cassava are detrimental due to its ability in ochratoxin A (OTA) production which carcinogenic and mutagenic to human. Objectives of this study were to isolate and identify Penicillium from cocoa, coffee beans and dried cassava in Yogyakarta by morphological and molecular characteristics, as well as to observe the ability of these isolates in OTA production on Yeast Extract Sucrose Agar (YES) medium. In this study, morphological characteristics were mainly based on the growth of isolates on identification media, while molecular characteristics were based on the similarity of PCR products using ITS4 and ITS5 as primers. OTA was detected by ELISA and UPLC methods. The result showed that 15 of 16 representative isolates obtained during this study were identified as Penicillium citrinum, one of the representative isolate from cocoa beans was identified as Penicillium paneum. Surprisingly, 13 among 15 of the obtained P. citrinum isolates from cocoa and coffee beans were positive in the production of OTA in YES medium, at the concentration of 4.64 to 25.26 µg/g media, while OTA was not detected in YES grown media by P. paneum and two isolates of P. citrinum from dried cassava. Conclusion of this study, the most found species Penicillium in cocoa and coffee beans were belong to P. citrinum which likely have a capability in the production of OTA.

A Convenient Method for Assessing Mycotoxin Production in Cultures of Aspergilli and Penicillia†

1996 ◽  
Vol 59 (6) ◽  
pp. 642-644 ◽  
Author(s):  
D. ABRAMSON ◽  
R. M. CLEAR
Keyword(s):  
Liquid Chromatography ◽  
Ochratoxin A ◽  
Human Exposure ◽  
Methylene Chloride ◽  
Penicillium Species ◽  
Extraction Solvent ◽  
Volatile Solvents ◽  
Culture Surface ◽  
Layer Chromatography ◽  
Yeast Extract Sucrose

Production of aflatoxins, sterigmatocystin, citrinin, and ochratoxin A by Aspergillus and Penicillium species on agar in 50-ml flasks after 21 days was detected and confirmed by thin-layer chromatography. Toxins were extracted with 2.5 ml of acidified methylene chloride for 20 to 60 min. Mycotoxin concentrations were estimated by liquid chromatography. Addition of formic acid to the methylene chloride extraction solvent effected an increase in recovery of ochratoxin A from Czapek-yeast (CY) agar from 20% to 91%. Recoveries of added mycotoxins were generally higher from CY agar than from yeast extract-sucrose (YES) agar and ranged from 11% (citrinin from YES agar) to 91% (ochratoxin A from CY agar). The method described is convenient, rapid, and reproducible, since it samples the entire culture surface. Human exposure to volatile solvents is also minimized.

scholarly journals Molecular characterization of lternaria solani isolates on tomato plant Lycopersicum esculentum Mill

2019 ◽  
pp. 379-400
Author(s):  
Hajer Mohammed Alsouyid ◽  
Nuria Ali Elamri ◽  
Haifa Mohamed Duzan ◽  
Abdunabi Mohamed Abughania ◽  
Ammar Khalifa Aslougi
Keyword(s):  
Significant Variation ◽  
Morphological Characteristics ◽  
Molecular Characteristics ◽  
Morphological Studies ◽  
Rapd Pcr ◽  
Pcr Products ◽  
Obvious Association ◽  
Group A ◽  
Group B ◽  
Group D

Early blight disease causes severe damage to the foliar part of solanaceous crops including tomato. Fifteen isolates (12 from tomato, 2 from potato, 1 from pepper) were collected from different sources in Tripoli. Field and laboratory studies were conducted to determine cultural behaviour on PDA medium, morphological, pathogenic and molecular variation between isolates tested. Colonies of isolates revealed variation in their cultural behaviour on PDA medium ranging from cottony to appressed growth, with colour ranging between light to dark olivaceous. The pigments released by the isolates changed the medium colour to grey or brown. Morphological studies of the fungal isolates exhibited short conidiophores bearing a single or chains of paired conidia. This study revealed a significant variation in conidial size for the isolates tested ranging from 23.45 to 46.90 x 7.70 to 14.00 µm. Pathogenicity testes on fruits, plants, and detached leaves of tomato indicated a high significant variation between isolates tested ranging from highly to moderate or weak pathogenic. Genetic diversity of A. solani isolates using RAPD-PCR with oligonuclotide primers revealed significant differences in the appearance of polymorphic and monomorphic banding patterns. Three primers (OPA-07, OPA-09, OPJ-09) out of ten were able to determine the genetic fingerprints of tested isolates. Cluster analysis of RAPD-PCR products showed that primer OPA-07 was able to classify the isolates into five groups: group A (TF4,TF7, TL1, TL3), group B (TF1,TF3, TF8, PEF), group C (TF5, TF6, TF9, POL1, POL2), whereas the remaining two isolates TL2 and TF2 were unique in their patterns and were designated as group U1 and U2 respectively. Primer OPA-09 revealed four distinct genetic groups designated as: group A (TF4, TF6, TF7, TF8, TL2), group B (POL1, TL3, TF3), group C (TF5, POL2) and group D (TL1, TF1, TF2, TF9, PEF). However primer OPJ-09 was able to split the isolates tested into four distinct clusters: group A (TF1, TF7, TL3, PEF), group B (TF3, TF8, POL2), group C (TF2, TF6, TF9, TL2) and group D (TF4, TF5). The results of RAPD-PCR demonstrate existence of considerable variation in molecular characteristics of A. solan iisolates. Accordingly these isolates were classified into different groups and unique patterns with no obvious association between the pattern of clustering of the isolates and their host of origin, morphological characteristics and pathogenicity.

scholarly journals Effect of relative humidity on biodeterioration of poultry feed and ochratoxin A production by Penicillium species

2016 ◽  
Vol 2 (1) ◽  
Author(s):  
V. Koteswara Rao ◽  
B. Aruna ◽  
Md. Rafiyuddin ◽  
K. Narasimha Rao ◽  
S. Girisham ◽  
...  
Keyword(s):  
Relative Humidity ◽  
Ochratoxin A ◽  
Poultry Feed ◽  
Penicillium Species

Effect of post-harvest treatments on the occurrence of ochratoxin A in raw cocoa beans

2015 ◽  
pp. 1-10 ◽  
Author(s):  
Kra Brou Didier Kedjebo ◽  
Tagro Simplice Guehi ◽  
Brou Kouakou ◽  
Noël Durand ◽  
Philippe Aguilar ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Cocoa Beans ◽  
Post Harvest

Measuring Ochratoxin A Concentrations in Coffee Beverages with Immunoaffinity Columns and Ultra-Performance Liquid Chromatography/Tandem Mass Spectrometry

2016 ◽  
Vol 99 (2) ◽  
pp. 469-474 ◽  
Author(s):  
Wen-Ling Chen ◽  
Chiung-Wen Chang ◽  
Chia-Yang Chen
Keyword(s):  
Ochratoxin A ◽  
Daily Intake ◽  
Ultra Performance Liquid Chromatography ◽  
Instant Coffee ◽  
Chain Stores ◽  
Chromatography Tandem Mass Spectrometry ◽  
Coffee Beans ◽  
Liquid Chromatography Tandem Mass ◽  
Core Column ◽  
Immunoaffinity Columns

Abstract This study developed and validated a method for measuring concentrations of ochratoxin A (OTA) in coffee beverages, not coffee beans. The new method involved extraction using immunoaffinity columns and ultra-performance LC (UPLC)-MS/MS using isotope-dilution techniques. The combination of a fused-core column and UPLC significantly shortened chromatographic time to 3 min compared to reported UPLC methods. The method was sensitive, with an LOD and LOQ of 0.52 and 1.73 pg/mL, respectively. Quantitative intraday (n = 4) and interday (n = 4) biases and RSD were both below 15%. The OTA levels in 40 samples of freshly brewed coffee from chain stores, 24 samples of canned ready-to-drink coffee, and 6 beverages made from instant coffee granules ranged from 1.60 to 93.2 pg/mL (90% positive), 6.00 to 131 pg/mL (100% positive), and 21.8 to 59.0 pg/mL (100% positive), respectively. Based on published tolerable daily intake, men and women in Taiwan should consume no more than 6.3 and 5.1 fifteen gram packages of instant coffee per day, respectively. Specific suggestions were not made for brewed coffee and canned coffee because of their large variation in OTA concentrations. This study should be more relevant to actual human exposure than those studying OTA in green, roasted, and ground coffee beans alone.

Genetic relationships among Brazilian strains of Aspergillus ochraceus based on RAPD and ITS sequences

2004 ◽  
Vol 50 (11) ◽  
pp. 985-988 ◽  
Author(s):  
Maria Helena Pelegrinelli Fungaro ◽  
Marciane Magnani ◽  
Laurival Antônio Vilas-Boas ◽  
Patrícia Cristina Vissotto ◽  
Márcia Cristina Furlaneto ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Sequence Data ◽  
Genetic Relationships ◽  
Its Region ◽  
Aspergillus Ochraceus ◽  
Coffee Bean ◽  
Coffee Beans ◽  
Major Species ◽  
Group A ◽  
Group B

Ochratoxin A (OA) is a mycotoxin that has been found in coffee beans and coffee beverages. Its toxicological profile includes carcinogenicity, nephrotoxicity, and immunotoxicity. Aspergillus ochraceus is the major species responsible for OA production in Brazilian coffee beans. The genetic relationships among 25 A. ochraceus strains collected from Brazilian coffee-bean samples were determined based on RAPD and internal transcribed spacer (ITS) sequence data. The isolates were resolved into 2 distinct groups, one with 4 strains (group A) and the other with 21 strains (group B). Specific nucleotide variations characterizing group A and B were found for both ITS1 and ITS2 regions. Group B is a new group proposed here to accommodate the majority of the Brazilian isolates. Each group was found to contain both toxigenic and nontoxigenic strains, indicating that there is no association between molecular genotypes and the ability to produce OA.Key words: Aspergillus ochraceus, ochratoxin A, ITS region (ITS1–5.8S–ITS2), RAPD.

scholarly journals Phylogenetic Relationships of Local Durian Species based on Morphological Characteristics and PCR-RFLP Analysis of the Ribosomal Internal Transcribed Spacer (ITS) DNA

2016 ◽  
Vol 8 (3) ◽  
pp. 362
Author(s):  
Fidia Fibriana ◽  
Lutfia Nur Hadiyanti
Keyword(s):  
Phylogenetic Relationships ◽  
Internal Transcribed Spacer ◽  
Morphological Characteristics ◽  
Biology Education ◽  
Rflp Analysis ◽  
Restriction Pattern ◽  
Internal Transcribed Spacers ◽  
Central Java ◽  
Pcr Rflp ◽  
Pcr Products

<p>In this study, twenty local durian accessions obtained from Central Java in situ collection were characterized using the morphological characteristics and the restriction patterns which generated from the region spanning the internal transcribed spacers ITS LEU and ITS 4. Morphological characteristics of durian leaf, stem, tree, and fruit showed variations for the different accessions, whereas polymerase chain reaction (PCR) products of ribosomal DNA region showed a low length of variation. The size of the PCR products and the restriction analyses with the restriction endonucleases Bsp1431yielded a restriction pattern for each accessions. The results of this study can be utilized by local durian farmers as a preliminary reference for durian propagation. The data obtained need to be supported by further research using the other molecular markers to obtain more accurate data. The clear identity of durian species can help the management of propagation systems by farmers to get superior local durian.</p><p><strong>How to Cite</strong></p><p>Fibriana, F., &amp; Hadiyanti, L. N. (2016). Phylogenetic Relationships of Local Durian Species based on Morphological Characteristics and PCR-RFLP Analysis of the Ribosomal Internal Transcribed Spacer (ITS) DNA. <em>Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(3), 362-370. </p>

Occurrence of ochratoxin A in cocoa beans: Effect of shelling

2005 ◽  
Vol 22 (6) ◽  
pp. 590-596 ◽  
Author(s):  
S Amézqueta ◽  
E González-Peñas ◽  
M Murillo ◽  
A López de Cerain
Keyword(s):  
Ochratoxin A ◽  
Cocoa Beans

Agrobacterium-mediated insertional mutagenesis of the ochratoxigenic fungus Aspergillus westerdijkiae

2007 ◽  
Vol 53 (1) ◽  
pp. 148-151 ◽  
Author(s):  
Marcia M Mata ◽  
Marta H Taniwaki ◽  
Beatriz T Iamanaka ◽  
Daniele Sartori ◽  
André L.M Oliveira ◽  
...  
Keyword(s):  
Ochratoxin A ◽  
Insertional Mutagenesis ◽  
Aspergillus Ochraceus ◽  
Agrobacterium Mediated Transformation ◽  
Morphological Variations ◽  
Carcinogenic Potential ◽  
Coffee Beans ◽  
Aspergillus Westerdijkiae ◽  
Polymerase Chain ◽  
First Time

Aspergillus westerdijkiae is a potent ochratoxin A (OTA) producer that has been found in coffee beans. OTA is known to have nephrotoxic effects and carcinogenic potential in animal species. Here we report for the first time the Agrobacterium-mediated transformation for Aspergillus westerdijkiae and the generation of ochratoxin-defective mutants. Conidia were transformed to hygromycin B resistance using strain AGL-1 of Agrobacterium tumefaciens. The obtained transformation frequency was up to 47 transformants per 106 target conidia. Among 600 transformants, approximately 5% showed morphological variations. Eight transformants with consistently reduced OTA production were obtained. Two of these transformants did not produce OTA (detection limit: 0.1 µg/kg); the other six mutants produced lower amounts of OTA (1%–32%) compared with the wild-type strain. By using thermal asymmetric interlaced polymerase chain reaction, we successfully identified a putative flavin adenine dinucleotide monooxygenase gene.Key words: Aspergillus ochraceus, Aspergillus westerdijkiae, Agrobacterium-mediated transformation, Agrobacterium-mediated insertional mutagenesis, ochratoxin A.

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