scholarly journals Detection Tuna and Processed Products Based Protein and DNA Barcoding

2015 ◽  
Vol 18 (2) ◽  
Author(s):  
Nuring Wulansari ◽  
Mala Nurilmala ◽  
N. Nurjanah
Keyword(s):  
Dna Barcoding ◽  
Pcr Amplification ◽  
Cyt B ◽  
High Demand ◽  
Fresh Fish ◽  
Meat Ball ◽  
Canned Tuna ◽  
Sds Page ◽  
Processed Products ◽  
Cyt B Gene

<p>Tuna is the second largest fishery commodity in Indonesia after the shrimp. Since the<br />high demand and the limited stock of tuna resulted in fraudulent chance. Authentication<br />is required to meassure consumers regarding the accuracy of its labeling and food<br />safety. In this study, the authentication was based on protein and DNA barcoding using<br />cytochrome-b gene (cyt-b) of the mitochondrial DNA as the target of gene. Primer of<br />cyt b gene was designed based on the tuna species. This study aimed to identify the<br />authenticity of tuna fresh and its processed products through protein using SDS-PAGE </p><p>and DNA barcoding techniques. The phases of this research were protein electrophoresis<br />by SDS-PAGE, DNA extraction, PCR amplification, electrophoresis and sequencing.<br />Samples of fresh fish (Tu1, Tu2, Tu3, Tu4, and Tu5) and processed tuna (canned and<br />steak) were successfully extracted. Result showed that SDS-PAGE proved the damage of<br />proteins in the processed tuna, so this method was not appropriate if it is used to identify<br />the authenticity of tuna. PCR electrophoresis results showed that the samples of tuna,<br />tuna steak, sushi, meat ball, abon, and caned tuna were successfully amplified in the range<br />of 500-750 bp except Ka3, which was in line with the target of DNA (620 bp). Resulted<br />sequences of Tu2, Tu3, Tu4 and Tu5 were identified according the results of morphometric<br />namely T. albacares, while Tu1 was identified as T. obesus with homology level of 99%.<br />Processed tunas (steak and canned tuna) were identified as T. albacares, as stated on the<br />labels.<br />Keywords: Authentication, cytb, DNA barcoding, design primer, SDS-PAGE</p>

scholarly journals Multiplex PCR Method of Detecting Pork to Guarantee Halal Status in Meat Processed Products

2019 ◽  
Vol 7 (3) ◽  
pp. 96-101
Author(s):  
M. Indriati ◽  
E. Yuniarsih
Keyword(s):  
Multiplex Pcr ◽  
Meat Product ◽  
Dna Amplification ◽  
Processed Meat ◽  
Target Sequence ◽  
Cyt B ◽  
Traditional Markets ◽  
Pcr Multiplex ◽  
Processed Products ◽  
Cyt B Gene

One of the halal parameters of food is must be free from pork among from the basic ingredients, additives and the manufacturing process. The aim of this study was ensure the halal status in the form of presence or absence of pork in processed meat product (meatballs and sausages) in traditional markets in Pandeglang Regency. PCR multiplex was a PCR technique that used several primers together in one reaction to amplify several target sequence. The genes often used as markers of animal or meat types include cytochrome b (cyt b), the existence of sequence variations in cyt b causes these genes are widely used as markers to distinguish material from different types of animals. The results showed that the cyt b gene proved successful in amplified DNA from beef and pork with different fragment lengths in the DNA mix of the 2 types of livestock in one reaction so that 2 DNA bands formed with different lengths according to the length of the fragment from each animal. From the results of research showed beef and pork control there was two fragments such as 274 bp for beef and 389 bp for pork. Multiplex PCR testing on meatball samples showed that the meatballs were tested 100% positive containing beef and 0% containing pork DNA. While testing on sausage products, there were one sausage brand that showed the results of DNA amplification of 398pb, which means the product was positive containing pork.

scholarly journals Morpho-molecular identification and first report of Fusarium equiseti in causing chilli wilt from Kashmir (Northern Himalayas)

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ammarah Hami ◽  
Rovidha S. Rasool ◽  
Nisar A. Khan ◽  
Sheikh Mansoor ◽  
Mudasir A. Mir ◽  
...  
Keyword(s):  
Dna Barcoding ◽  
Pcr Amplification ◽  
Its Region ◽  
Fungal Species ◽  
Pathogenicity Test ◽  
Kashmir Valley ◽  
Fusarium Equiseti ◽  
Infected Root ◽  
Fusarium Sp ◽  
High Degree

AbstractChilli (Capsicum annuum L.) is one of the most significant vegetable and spice crop. Wilt caused by Fusarium Sp. has emerged as a serious problem in chilli production. Internal transcribed spacer (ITS) region is widely used as a DNA barcoding marker to characterize the diversity and composition of Fusarium communities. ITS regions are heavily used in both molecular methods and ecological studies of fungi, because of its high degree of interspecific variability, conserved primer sites and multiple copy nature in the genome. In the present study we focused on morphological and molecular characterization of pathogen causing chilli wilt. Chilli plants were collected from four districts of Kashmir valley of Himalayan region. Pathogens were isolated from infected root and stem of the plants. Isolated pathogens were subjected to DNA extraction and PCR amplification. The amplified product was sequenced and three different wilt causing fungal isolates were obtained which are reported in the current investigation. In addition to Fusarium oxysporum and Fusarium solani, a new fungal species was found in association with the chilli wilt in Kashmir valley viz., Fusarium equiseti that has never been reported before from this region. The studies were confirmed by pathogenicity test and re-confirmation by DNA barcoding.

scholarly journals Genetic characterization and phylogenetic study of Indonesian indigenous catfish based on mitochondrial cytochrome B gene

2020 ◽  
Vol 13 (1) ◽  
pp. 96-103
Author(s):  
Dorothea Vera Megarani ◽  
Herjuno Ari Nugroho ◽  
Zahrah Prawita Andarini ◽  
Yura Dwi Risa B. R. Surbakti ◽  
Rini Widayanti
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Cytochrome B ◽  
Genetic Characterization ◽  
Phylogenetic Study ◽  
Cyt B ◽  
Phylogenetic Structure ◽  
Mitochondrial Cytochrome B ◽  
Sperata Seenghala ◽  
Cyt B Gene

Aim: This study aimed to determine the genetic characterization and phylogenetic structure of Indonesian indigenous catfish using cytochrome B (Cyt B) sequences. Materials and Methods: The genomes of 26 catfishes caught from nine rivers from nine different geographical locations around Indonesia were analyzed. The tissue isolation method was used to isolate the total genome of the fishes. Furthermore, polymerase chain reaction was done to amplify the mtDNA Cyt B using the CytBF and CytBR primers. Following sequencing, the analysis of genetic variation and the phylogenetic relationship was performed using MEGA version X software. Results: Cyt B gene sequencing attained a total of 1139 nucleotides encrypting 379 amino acids for all samples. The ClustalW alignment program using MEGA X software revealed 395 substituted nucleotides, which then translated into 63 amino acid variation sites among all 26 samples. No amino acids in catfish BB were different compared to catfish PM, MP, and KR2,3. Catfish MS had one modified amino acid; KR1 and KS had two different amino acids; BF had 38 different amino acids; EM had 31 different amino acids; and BSBJ had 26 different amino acids compared to catfish BB. The most significant alteration of amino acids was between catfish EM and BF (49 amino acids). Conclusion: Indonesian catfish were divided into five clades based on the Cyt B gene. Samples KR and MP (Sumatra); MS and BB (Kalimantan); and PM (Java) were clustered with Hemibagrus nemurus and Hemibagrus wyckioides (Bagridae family). Samples from Kalimantan (KS) and one sample of KR (KR1) from Sumatra were clustered with Sperata seenghala and Hemibagrus spilopterus (Bagridae family). Samples from Java (BSBJ) were clustered with Pseudolais pleurotaenia (Pangasiidae family). Samples EM (Java) were together with Mystus cavasius (Bagridae family). Samples from West Papua were clustered with Potamosilurus latirostris (Ariidae family).

scholarly journals Kajian Penanda Genetik Gen Sitokrom b DNA Mitokondria Ikan Lais dari Sungai Kampar Riau

2018 ◽  
Vol 10 (1) ◽  
pp. 6
Author(s):  
Roza Elvyra ◽  
Dedy Duryadi Solihin
Keyword(s):  
Species Diversity ◽  
Molecular Marker ◽  
Phylogenetic Relationship ◽  
Cytochrome B ◽  
Mitochondrial Cytochrome ◽  
Cyt B ◽  
Phylogenetic Marker ◽  
Mitochondrial Cytochrome B ◽  
Cyt B Gene

The mitochondrial cytochrome b (cyt-b) gene as a phylogenetic marker of lais fish Kryptopterus schilbeides from Kampar River in Riau has been studied. This is a prelimininary research on the utility of cyt-b gene as a molecular marker to obtain species diversity and phylogenetic relationship among Kryptopterus fishes from Kampar River. The primers of L14841 and H15149 were used to amplify the cyt-b gene. The results showed that K. schilbeides has isoleusine at site-81 and metionine at site-114; K. schilbeides from Kampar River and K. schilbeides from GenBank form a phylogeny cluster at 45% value.

scholarly journals Microbiological and Nutritional Properties of Frankfurter-Type Fish Sausage

2018 ◽  
pp. 27-34
Author(s):  
Nkrumah T ◽  
Akwetey WY
Keyword(s):  
Nutritive Value ◽  
Frozen Storage ◽  
Nutritional Composition ◽  
Negative Effects ◽  
Nutritional Values ◽  
Fresh Fish ◽  
E Coli ◽  
Fish Meat ◽  
Preservation Technology ◽  
Processed Products

Fish meat is a common and broadly used food due to its high nutritional values yet the bones and flavour of fresh fish can be unpleasant. The unpleasant fishy flavour makes it difficult to handle and process. The fishy flavour is as a result of microbial presence and growth. Studies associated with microbial contaminations have concentrated more on the carcass. Such contaminations which affect the microbiological status of processed products can emanate from spices and other non-meat ingredients, environment, equipment and handlers. The successful application of processing/preservation technology results in the conservation of desirable qualities in stabilized and varietal fish products. This study sought to use fish in the manufacture of frankfurter-type sausages, which could have improved preservation characteristics without any adverse effects on sensory properties. The nutritional composition and microbiological safety of fresh fish and sausages were determined using the methods described by AOAC and ICMSF respectively. The study showed that, catfish sausages were higher in protein (15.69 %) and were lower in fat (10.66%) compared to the other sausages. Total Viable Counts (TVC) were within the accepted limits (106 and 107cfu/g) for fish and pork respectively. E. coli was not detected in any of the treatments during frozen storage for 6 weeks. It was concluded that catfish frankfurter has high nutritive value because it contained less fat but with higher crude protein. Like pork frankfurters, both catfish and mackerel sausages could be stored for six weeks without any negative effects on microbial quality.

Histamine in Fish Products Randomly Collected in Southern Italy: A 6-Year Study

2020 ◽  
Vol 83 (2) ◽  
pp. 241-248 ◽  
Author(s):  
ANTONELLO CICERO ◽  
GAETANO CAMMILLERI ◽  
FRANCESCO GIUSEPPE GALLUZZO ◽  
ILARIA CALABRESE ◽  
ANDREA PULVIRENTI ◽  
...  
Keyword(s):  
Southern Italy ◽  
Street Vendors ◽  
Mean Values ◽  
Fresh Fish ◽  
Smoked Fish ◽  
Canned Tuna ◽  
Fish Samples ◽  
Fish Product ◽  
Array Detection ◽  
Ultrahigh Performance Liquid Chromatography

ABSTRACT In total, 4,615 fresh and processed fish samples collected from 2010 to 2015 were analyzed for histamine by ultrahigh-performance liquid chromatography with diode array detection. Histamine levels were detected in 352 (7.6%) samples, with a maximum of 4,110 mg kg−1 and mean values of 908.9 ± 1,226.79 and 344.01 ± 451.18 mg kg−1 for fresh and processed fish samples, respectively. No histamine levels were found in canned tuna and smoked fish samples in contrast to most of the data reported in the literature. A low percentage (2.79%) of noncompliant samples was found. The highest mean values were found during 2011 and 2015 for fresh and processed fish samples, respectively, showing a significant (P &lt; 0.05) difference between the sampling years. The histamine contents found in fresh fish samples were significantly higher (P &lt; 0.05) than those of processed samples. Most of the positive samples came from street vendors, suggesting the need to improve inspection measures in these commercial categories to ensure fish product safety. HIGHLIGHTS

scholarly journals Partial sequence analysis of mitochondrial cytochrome B gene of Labeo calbasu of Bangladesh

2019 ◽  
Vol 5 (1) ◽  
pp. 25-30
Author(s):  
RA Begum ◽  
MT Alam ◽  
H Jahan ◽  
MS Alam
Keyword(s):  
Genetic Diversity ◽  
Cytochrome B ◽  
Tissue Sample ◽  
Sequence Similarity ◽  
Cytochrome B Gene ◽  
Mitochondrial Cytochrome ◽  
Cyt B ◽  
Multiple Sequence ◽  
Mitochondrial Cytochrome B ◽  
Cyt B Gene

Labeo calbasu (Family Cyprinidae) was studied at DNA level to know genetic diversity within and between species. The mitochondrial cytochrome b (cyt-b) gene of L. calbasu was sequenced and compared to the corresponding sequences of other Labeo species. DNA was isolated from the tissue sample of L. calbasu using phenol: chloroform extraction method. Forward and reverse primers were designed to amplify the target region of cytochrome b gene. A standard PCR protocol was used for the amplification of the desired region. Then, the forward and reverse sequences obtained were aligned and edited to finalize a length of 510 nucleotides which was submitted to NCBI genbank database. Nucleotide BLAST of this sequence at NCBI resulted 100% sequence similarity with L. calbasu sequence of the same region of cyt-b gene. Multiple sequence alignment of the sequence with seven more Labeo species sequences revealed 120 polymorphic sites, which have been mark of diversity among the species and might be used in molecular identification of the Labeo species. A constructed phylogenetic tree has shown relationship among the Labeo species. This research demonstrated the usefulness of mitochondrial DNA-based approach in species identification. Further, the data will provide appropriate background for studying genetic diversity within-species of the Labeo species in general and of L. calbasu in particular. J. Biodivers. Conserv. Bioresour. Manag. 2019, 5(1): 25-30

Alternaria alternata (Fr.) Keissl with mutation G143A in the Cyt b gene is the source of a difficult-to-control allergen

2017 ◽  
Vol 25 (1) ◽  
pp. 469-478
Author(s):  
Adrian Duba ◽  
Klaudia Goriewa ◽  
Urszula Wachowska ◽  
Marian Wiwart
Keyword(s):  
Alternaria Alternata ◽  
Cyt B ◽  
Cyt B Gene

scholarly journals Allele-Specific PCR for the Detection of Azoxystrobin Resistance in Didymella bryoniae

Plant Disease ◽  
2014 ◽  
Vol 98 (12) ◽  
pp. 1681-1684 ◽  
Author(s):  
Mavis J. Finger ◽  
Venkatesan Parkunan ◽  
Pingsheng Ji ◽  
Katherine L. Stevenson
Keyword(s):  
Dna Sequences ◽  
Cyt B ◽  
Didymella Bryoniae ◽  
Specific Pcr ◽  
Pcr Product ◽  
Allele Specific ◽  
Polymerase Chain ◽  
Sensitive Allele ◽  
Cyt B Gene ◽  
Allele Specific Pcr

Gummy stem blight (GSB), caused by the fungus Didymella bryoniae, is considered the most widespread and destructive disease of watermelon in the southeastern United States. The quinone outside-inhibiting (QoI) fungicide azoxystrobin (AZO), which inhibits mitochondrial respiration by binding to the outer, quinone-oxidizing pocket of the cytochrome bc1 (cyt b) enzyme complex, was initially very effective in controlling GSB. However, resistance to AZO has been observed in D. bryoniae in many watermelon-producing regions. In this study, the DNA sequences of partial cyt b genes of four AZO-resistant (AZO-R) and four AZO-sensitive (AZO-S) isolates of D. bryoniae confirmed the amino acid substitution of glycine by alanine at the 143 codon (G143A) in the AZO-R isolates tested. Allele-specific primers were designed to detect the resistant or sensitive allele at codon 143 of the cyt b gene, which amplified a 165-bp polymerase chain reaction (PCR) product from genomic DNA of nine AZO-R and nine AZO-S isolates of D. bryoniae, respectively. The primer pairs did not amplify DNA from other pathogens tested in the study. The results indicated that the PCR assays developed in the study were specific in differentiating AZO-R and AZO-S isolates and could facilitate AZO resistance detection in D. bryoniae.

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