scholarly journals The Quality of DNA Isolated from Processed Food and Feed via Different Extraction Procedures

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1188 ◽  
Author(s):  
Zora Piskata ◽  
Eliska Servusova ◽  
Vladimir Babak ◽  
Michaela Nesvadbova ◽  
Gabriela Borilova
Keyword(s):  
Meat Products ◽  
Pcr Amplification ◽  
Single Species ◽  
Extraction Methods ◽  
Optimal Choice ◽  
Pcr Analysis ◽  
Processed Food ◽  
Extraction Procedures ◽  
Heat Treated ◽  
Food And Feed

The extraction of DNA is a critical step for species identification by PCR analysis in processed food and feed products. In this study, eight DNA extraction procedures were compared—DNeasy Blood and Tissue Kit, DNeasy mericon Food Kit, chemagic DNA Tissue 10 Kit, Food DNA Isolation Kit, UltraPrep Genomic DNA Food Mini Prep Kit, High Pure PCR Template Preparation Kit, phenol—chloroform extraction, and NucleoSpin Food—Using self-prepared samples from both raw and heat-processed and/or mechanically treated muscles and different types of meat products and pet food (pork, beef, and chicken). The yield, purity, and suitability of DNA for PCR amplification was evaluated. Additionally, comparisons between the effectiveness of various extraction methods were made with regard to price, and labor- and time-intensiveness. It was found that the DNeasy mericon Food Kit was the optimal choice for the extraction of DNA from raw muscle, heat-treated muscle, and homemade meat products from multiple and single species.

Detection of Pork in Heated Meat Products by the Polymerase Chain Reaction

1994 ◽  
Vol 77 (3) ◽  
pp. 617-622 ◽  
Author(s):  
Rolf Meyer ◽  
Urs Candrian ◽  
Jürg Lüthy
Keyword(s):  
Polymerase Chain Reaction ◽  
Meat Products ◽  
Ribosomal Gene ◽  
Pcr Analysis ◽  
Growth Hormone Gene ◽  
Chain Reaction ◽  
Immunological Tests ◽  
Heat Treated ◽  
Polymerase Chain ◽  
Meat Samples

Abstract A new method for the specific, sensitive, and semiquantitative detection of pork (Sus scrota) in heat-treated meat products has been developed. DNA was isolated from meat samples by using a DNA-binding resin and subjected to polymerase chain reaction (PCR) analysis. First, oligonu-cleotides yielding a specific 137-base-pair (bp) fragment from eucaryotic DNA amplified from a highly conserved region of the 18-S ribosomal gene was used to assess DNA quality. Second, the presence of pork DNA was determined with specific oligonu-cleotides yielding a 108-bp fragment amplified from the porcine growth hormone gene. The test detected pork in fresh or heated meat mixtures of pork in beef at levels below 2%. This approach was superior to commercially available immunological tests that were not able to detect levels of pork less than 20% in cooked meat or less than 10% in fresh meat.

scholarly journals Selection of the most suitable method for the extraction of DNA from foods and feeds for species identification

2010 ◽  
Vol 58 (2) ◽  
pp. 169-174
Author(s):  
Michaela Nesvadbová ◽  
Aleš Knoll ◽  
Anna Vašátková
Keyword(s):  
Species Identification ◽  
Genomic Dna ◽  
Pcr Amplification ◽  
Degraded Dna ◽  
Dna Purification ◽  
Pcr Analysis ◽  
Isolation System ◽  
Dna Yield ◽  
Pcr Products ◽  
Food And Feed

High quality and purity of DNA isolated from food and feed is essential for species identification and has unpredictable influences an effect of analysis. In this study, the efficiency of eight different methods for DNA isolation was investigated. For DNA extraction, the raw chicken meet, ham, sausages, tinned lunch meat, pate, tinned feeds for dogs, complete granulated feeds for dogs and chicken flour were used. Kits of several different producers, i.e.: NucleoSpin Food (Marchery-Nagel), Wizard Genomic DNA Purification Kit (Promega), Invisorb Spin Food Kit I (Invitek), Wizard SV Genomic DNA Purification System (Promega), JetQuick Tissue DNA Spin Kit (Genomed), RNA Blue (Top-Bio), JetQuick Blood & Cell Culture Kit (Genomed), QIAamp DNA Mini Kit and QIAamp DNA Blood Mini Kit (Qiagen) were employed in the study. Gel agarose electrophoresis for primary verification of DNA quality was performed. The isolates were subsequently assessed for quantity and quality using by spectrophotometer Nanodrop 2000 (Thermo Scientific). To verify of template usability and quality of isolated DNA, the polymerase chain reaction (PCR) was used.Differences between isolated DNA from tinned products and meat, ham, sausage, granulated dog feed and chicken flour were found. In tinned food and feed, the DNA was more degraded, DNA content and DNA purity was lower and also PCR amplification was the most difficult. Overall DNA yield and quality have important influence on PCR products amplification. The best results were obtained with NucleoSpin Food and JetQuick Tissue DNA Spin Kit. DNA extracted by these methods proved highest yields, purity and template quality in all foods and feeds and the results of PCR analysis are excellent reproducible. Analyses showed that results depended on different food or feed using and dif­fe­rent isolation system.The results of this work will be utilized to choose the suitable isolating kit for educational course, which is designed for students and also for following research and analyses.

Rapid quantification of Salmonella Typhimurium inoculated to meat products by real-time PCR

2009 ◽  
Vol 57 (1) ◽  
pp. 25-38 ◽  
Author(s):  
Ching-Yang Cheng ◽  
Jing-Ruei Chi ◽  
Sin-Rong Lin ◽  
Chi-Chiang Chou ◽  
Chin-Cheng Huang
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Pcr Amplification ◽  
Food Samples ◽  
Pcr Analysis ◽  
Target Cells ◽  
Chicken Nuggets ◽  
Reliable Technique ◽  
Isolation And Purification

The objective of this study was to use a 5′-nuclease (TaqMan) real-time PCR method with primers and probe specific to thespaQgene as a rapid approach to quantitatively determineSalmonellaTyphimurium. The result showed that the correlation coefficient between real-time PCR estimates and bovine serum albumin (BSA) plate counts ofS. Typhimurium was 0.99, independently of 105-fold numbers of bystanderEscherichia coliO157:H7 or total viable counts. The sensitivity of the real-time quantitative PCR assay was 10 CFU/mL for pureS. Typhimurium culture without enrichment. A known number ofS. Typhimurium target cells were inoculated to dumpling fillings and chicken nuggets and DNA was extracted for real-time PCR analysis. The sensitivity was 60 CFU/g forS. Typhimurium inoculated to the food samples without any preceding procedure of enrichment. The duration of the entire experiment from DNA isolation and purification to PCR amplification was less than 12 h. This study demonstrated that realtime PCR is a rapid and reliable technique for quantifyingS. Typhimurium possessing thespaQgene in pure culture and in meat products.

scholarly journals Suitability of different tissue fixatives for subsequent PCR analysis of Cysticercus ovis

2012 ◽  
Vol 49 (2) ◽  
pp. 67-70 ◽  
Author(s):  
M. Kolesárová ◽  
R. Herich ◽  
M. Levkut ◽  
J. Čurlík ◽  
M. Levkut
Keyword(s):  
Retrospective Studies ◽  
Pcr Amplification ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Fresh Tissue ◽  
Carnoy’S Solution ◽  
Negative Results ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Cysticercus Ovis

AbstractPCR amplification of specific DNA regions is a powerful tool for retrospective studies, but not all preservation or fixation methods render DNA that is suitable for subsequent amplification. Several factors affect sensitivity of polymerase chain reaction (PCR) amplification. There were reported the effects of commonly used fixation solutions — 10 % neutral buffered formalin, 20 % neutral buffered formalin and Carnoy’s solution and the efficiency of PCR amplification in fresh tissue and paraffin (or wax) embedded samples of Cysticercus ovis. DNA from samples was isolated and PCR product of 1300 bp was amplified. Results indicated that the samples fixed in Carnoy’s solution produced reliable amplification of desired fragments. The samples that were fixed in 10 % and 20 % neutral buffered formalin brought negative results.

scholarly journals Salmonella spp. in RASFF notifications involving Croatia in the period from 01/01/2014 to 31/12/2018

2019 ◽  
Vol 2 (5-6) ◽  
pp. 24-36
Author(s):  
Emilija Friganović ◽  
Nikolina Tokmakčija ◽  
Ančica Sečan Matijaščić ◽  
Mirko Kelava ◽  
Mladenka Šarolić ◽  
...  
Keyword(s):  
Meat Products ◽  
Product Category ◽  
Food Products ◽  
Poultry Meat ◽  
Alert System ◽  
Salmonella Spp ◽  
Fast Exchange ◽  
Food And Feed ◽  
Foodborne Outbreaks ◽  
Rapid Alert System

The Rapid Alert System for Food and Feed (RASFF) enables a fast exchange of information between bodies and institutions involved in the system (EU Member State national food safety authorities, Commission, EFSA, ESA, Norway, Liechtenstein, Iceland and Switzerland) in order to respond promptly to the health risks associated with food and feed. Salmonella is an important cause of EU foodborne outbreaks, most frequently reported pathogenic microorganism in food in the last few years. The aim of this study was to analyze RASFF notifications on food products contaminated with Salmonella spp. involving Croatia in the period from 01/01/2014 to 31/12/2018. All data were downloaded from the RASFF database (RASFF portal) and processed in MS Excel 2010. The collected data provided information on the: country(ies) of origin and distribution of the contaminated product, notifying country, product and product category, notification type, risk decision, action taken, distribution status and, for some incidents, a Salmonella spp. serovar. Notifications mainly concerned "poultry meat and poultry meat products". Just over half of the reported food products originated from Poland, Brazil and Italy. Croatia was notifying country in almost half of the published notifications. Majority of the notifications were classified as alert notifications and of serious risk. Most of the Salmonella spp. notifications were based on official controls on the market and on company's own check.

scholarly journals Molecular identification of four Sarcocystis species in cattle from Lithuania, including S. hominis, and development of a rapid molecular detection method

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Petras Prakas ◽  
Živilė Strazdaitė-Žielienė ◽  
Vytautas Januškevičius ◽  
Francesco Chiesa ◽  
Agnė Baranauskaitė ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Light Microscope ◽  
Bos Taurus ◽  
Geographical Location ◽  
Single Species ◽  
Diaphragm Muscle ◽  
Common Finding ◽  
Pcr Analysis ◽  
Sarcocystis Species ◽  
Specific Pcr

Abstract Background Six Sarcocystis species are known to use cattle (Bos taurus) as the intermediate host, two of which, S. hominis and S. heydorni, are zoonotic. There is a need for a method that will enable rapid identification of the Sarcocystis species in cattle. Methods The diaphragm muscles of 102 cattle from Lithuania were examined for the presence of Sarcocystis spp., using two different methods for species identification. Individual sarcocysts were isolated from squash preparations of the diaphragm muscle under the light microscope, followed by genetic characterisation of excised cysts using sequence analysis of the 18S rRNA (18S rRNA) and cytochrome c oxidase subunit I (cox1) genes. The same cattle muscle samples were digested and species-specific PCR analyses targeting cox1 were developed to identify the Sarcocystis isolates to the species level. Results Under the light microscope, sarcocysts were detected in 87.3% of animals, and Sarcocystis infection was verified in all digested samples. Three species, namely S. cruzi (n = 20), S. bovifelis (n = 23) and S. hirsuta (n = 6), were identified by DNA sequence analysis of isolated sarcocysts. Based on sequence analysis of cox1, the level of genetic variability depended on Sarcocystis species and geographical location. Four Sarcocystis species, S. cruzi (96.1%), S. bovifelis (71.6%), S. hirsuta (30.4%) and S. hominis (13.7%), were confirmed in the digested samples. In individual samples, the most common finding was two species of Sarcocystis (44.1%), followed by three species (26.5%), a single species (24.5%) and four species (4.9%). Conclusions Although examination of tissue preparations under the light microscrope did not detect any sarcocysts belonging to S. hominis, this species was identified in the digested samples subjected to a cox1-specific PCR analysis. These results demonstrate the need for effective molecular diagnosis techniques to detect Sarcocystis spp., which may be present at a lower prevalence and not detectable among the limited number of sarcocysts identified individually under the light microscope.

scholarly journals Outbreak of trichinellosis in eastern Slovakia

2009 ◽  
Vol 46 (4) ◽  
pp. 209-213 ◽  
Author(s):  
Z. Paraličová ◽  
J. Kinčeková ◽  
I. Schréter ◽  
P. Jarčuška ◽  
P. Dubinský ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Meat Products ◽  
Meat Processing ◽  
Igg Antibodies ◽  
Raw Meat ◽  
Laboratory Parameters ◽  
Reactive Protein ◽  
Heat Treated ◽  
Periorbital Oedema ◽  
Laboratory Changes

AbstractTrichinellosis is a zoonosis caused by ingestion of undercooked raw meat from animals that harbour infectious larvae. In most of the Slovak regions there is ongoing life cycle of circulating trichinellosis in wild carnivores and wild boar population. The outbreak of trichinellosis occured in Rožňava district east Slovakia during spring in 2008. Ten members of farmer’s family and their relatives got ill while processing meat from home-made pig-slaughter for meals and meat products intended for wedding dinner. During the meat processing all of them tasted raw meat. Moreover, another 45 persons were exposed to this infection by eating heat-treated meat products. The most common predominant clinical signs were: myalgias, fever, fatigue, exanthema and periorbital oedema. On the 40th day after infection there were intermediate to high titres of trichinella IgG antibodies detected (10 patients), high levels of eosinophilia (10 patients) with maximum of 6.76 × 109/l (55 %) and profound changes in selected laboratory parameters: decreased levels of total proteins, increased levels of alpha 1-globulin and C reactive protein. Presence of IgG antibodies as well as aforementioned laboratory parameters was important markers of trichinellosis in our study, whereas other laboratory changes (leukocytosis, high levels of activity lactate dehydrogenase and creatine kinase) were detected only in few hospitalized patients.

Determination of Histamine in Japanese Spanish Mackerel (Scomberomorus niphonius) Meat Implicated in a Foodborne Poisoning

2019 ◽  
Vol 82 (10) ◽  
pp. 1643-1649 ◽  
Author(s):  
CHIU-CHU HWANG ◽  
PEI-HUI TSENG ◽  
YI-CHEN LEE ◽  
HSIEN-FENG KUNG ◽  
CHUN-YUNG HUANG ◽  
...  
Keyword(s):  
Meat Products ◽  
Pcr Amplification ◽  
Enterobacter Aerogenes ◽  
Potential Hazard ◽  
Bacterial Strains ◽  
Spanish Mackerel ◽  
Rdna Sequencing ◽  
Fish Samples ◽  
Raw Fish ◽  
Scomberomorus Niphonius

ABSTRACT An incident of foodborne poisoning causing illness in seven victims due to ingestion of fried Japanese Spanish mackerel (JS mackerel; Scomberomorus niphonius) meat occurred in September 2014 in Hualien County, eastern Taiwan. Of the two suspected fish meats, one raw sample contained 3,318 ppm of histamine and one fried sample contained 1,906 ppm of histamine, levels which are greater than the potential hazard action level (500 ppm) in most illness cases. Given the allergy-like symptoms of the victims and the high histamine content in the suspected fish samples, this foodborne poisoning was strongly suspected to be caused by histamine intoxication. In addition, five histamine-producing bacterial strains isolated from suspected raw fish samples, capable of producing 152 to 1,020 ppm of histamine in Trypticase soy broth supplemented with 1.0% l-histidine, were identified as Hafnia alvei (one strain), Enterobacter aerogenes (two strains), Raoultella ornithinolytica (one strain), and Morganella morganii (one strain) by 16S rDNA sequencing with PCR amplification. Moreover, 12 raw fish samples and 39 fried fish samples from retail stores were collected and tested to determine the occurrence of histamine. Two of 12 commercial raw fish samples (16.7%) had histamine levels greater than the U.S. Food and Drug Administration guideline for decomposition of 50 ppm for scombroid fish or product or a combination of both. To our knowledge, this is the first report in Taiwan to demonstrate that the JS mackerel meat products could cause histamine intoxication.

Sign in / Sign up

Export Citation Format

Share Document