Survival of Salmonellae in Pasteurized, Refrigerated Calcium-Fortified Orange Juice

2001 ◽  
Vol 64 (9) ◽  
pp. 1299-1304 ◽  
Author(s):  
MANAN SHARMA ◽  
LARRY R. BEUCHAT ◽  
MICHAEL P. DOYLE ◽  
JINRU CHEN
Keyword(s):  
Polymerase Chain Reaction ◽  
Orange Juice ◽  
Storage Period ◽  
Animal Origin ◽  
Calcium Citrate ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Acidic Environments ◽  
Number Of Cells ◽  
Salmonella Heidelberg

Studies were done to determine the survival of salmonellae in orange juice as affected by fortification with calcium. Four brands of commercially pasteurized orange juice fortified with calcium (350 mg/240-ml serving) and nonfortified juice were inoculated separately with three types of inocula: strains of Salmonella Muenchen (inoculum 1), serotypes of human and animal origin (inoculum 2), and isolates from raw produce- and juice-associated outbreaks (inoculum 3). Juice inoculated with populations of 6.6 to 7.0 log10 CFU of Salmonella per ml was held at 4°C for up to 32 days. The number of cells of inoculum 1 that survived in juice fortified with calcium lactate/tricalcium phosphate (CaL/TCP) was significantly lower (P ≤ 0.05) (2.80 log10 CFU/ml) than in nonfortified juice (3.50 log10 CFU/ml) after 32 days' storage. Death of salmonellae in inocula 1 and 2 was less in juice fortified with TCP (3.21 and 3.33 log10 CFU/ml, respectively) than in the nonfortified juice (3.75 and 4.15 log10 CFU/ml, respectively). During the 32-day storage period, populations in inocula 1 and 3 showed significantly less inactivation (2.62 and 3.12 log10 CFU/ml, respectively) in juice fortified with calcium citrate (CC) than in nonfortified juice (3.14 and 3.60 log10 CFU/ml, respectively).There were no significant differences in the survival of Salmonella in juice fortified with calcium citrate malate (CCM) and nonfortified juice. Polymerase chain reaction (PCR) typing of randomly selected Salmonella colonies revealed that Salmonella Heidelberg in inoculum 2 and Salmonella Baildon and Salmonella Poona in inoculum 3 were the most prevalent at the end of the 32-day storage period at 4°C, suggesting that serotypes selected for use in inocula differed in tolerance to acidic environments. This study reveals that the form of calcium used to fortify orange juice may affect the survival of Salmonella.

scholarly journals Evaluation of a Polymerase Chain Reaction-Based Heteroduplex Assay for Detecting the Adulteration of Processed Orange Juice with Mandarin Juice

2006 ◽  
Vol 89 (4) ◽  
pp. 1052-1060 ◽  
Author(s):  
Rachel Mooney ◽  
Louise Chappell ◽  
Angus I Knight
Keyword(s):  
Polymerase Chain Reaction ◽  
Orange Juice ◽  
Polyacrylamide Gel Electrophoresis ◽  
Pcr Amplification ◽  
Intergenic Spacer ◽  
Target Sequence ◽  
Chain Reaction ◽  
Good Repeatability ◽  
Polymerase Chain ◽  
Mandarin Juice

Abstract A polymerase chain reaction (PCR)-based heteroduplex assay was evaluated for the detection of mandarin juice in processed orange juice. PCR amplification of a fragment of the chloroplast trnT-trnL intergenic spacer derived from mixtures of DNA extracted from orange and mandarin juice resulted in heteroduplex formation. The heteroduplex resulted from the co-amplification of a fragment containing an 8 base-pair indel that distinguished mixtures of orange and mandarin juice from orange juice and mandarin juice alone. The heteroduplex assay was evaluated against authentic juices obtained from different citrus species and confirmed that the marker was homogeneous within Citrus. The data obtained demonstrated maternal inheritance of chloroplast type in Citrus sp. and allowed the identification and confirmation of the maternal parentage of unknown and known citrus hybrids. Analysis of the quantitative potential of the PCR and polyacrylamide gel electrophoresis (PAGE) analysis demonstrated good repeatability with a coefficient of variation of 7.5%. Greatest sources of variance in experimental results were attributable to species and varietal differences in the levels of the PCR target. Mandarin juice contained approximately 18% (w/v) less PCR target sequence than did orange juice. The assay was tested in a blind trial using processed juices and correctly identified 20/22 samples with no false-positive results.

Application of polymerase chain reaction for detection of goats' milk adulteration by milk of cow

2001 ◽  
Vol 68 (2) ◽  
pp. 333-336 ◽  
Author(s):  
JACEK BANIA ◽  
MACIEJ UGORSKI ◽  
ANTONI POLANOWSKI ◽  
ERYK ADAMCZYK
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Analysis ◽  
Marker Gene ◽  
Meat Products ◽  
Universal Primers ◽  
Animal Origin ◽  
Chain Reaction ◽  
Specific Primers ◽  
Single Stranded Conformational Polymorphism ◽  
Polymerase Chain

Numerous methods based on DNA analysis have been employed in the food industry to monitor adulterations of food products of animal origin. Among them the most frequently used are: polymerase chain reaction (PCR) amplification of a marker gene fragment(s) with universal primers, or amplification of DNA with species-specific primers. PCR-products of different origin can be discriminated by size, restriction fragment length polymorphism (RFLP) or single stranded conformational polymorphism (SSCP) analysis. These methods have been used for identification, and differentiation between, the animal origins of raw or heat-treated meat and meat products (Chikuni et al. 1994; Meyer et al. 1994, 1995; Zehner et al. 1998; Behrens et al. 1999; Guoli et al. 1999; Hopwood et al. 1999; Matsunaga et al. 1999; Wolf et al. 1999). These approaches are also applicable to the analysis of dairy products. However, adulterations of goats' milk and its products are traditionally tested by immunological and/or electrophoretic methods (Amigo et al. 1992; Levieux & Venien, 1994; Mimmo & Pagani, 1998). So far, only a few DNA-based techniques designed to detect the presence of bovine DNA in goats' milk have been described (Plath et al. 1997; Branciari et al. 2000). This paper presents a one-step PCR procedure for detection of adulteration of goats' milk with cows' milk. The method, employing bovine-specific primers for amplification of a 274 bp fragment of cytochrome b DNA, seems to be simple, fast, specific and sensitive.

Use of duplex polymerase chain reaction (duplex-PCR) technique to identify bovine and water buffalo milk used in making mozzarella cheese

2001 ◽  
Vol 68 (4) ◽  
pp. 689-698 ◽  
Author(s):  
STEFANO REA ◽  
KOICHI CHIKUNI ◽  
RAFFAELLA BRANCIARI ◽  
RAM SUKASI SANGAMAYYA ◽  
DAVID RANUCCI ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Species Identification ◽  
Water Buffalo ◽  
Bovine Milk ◽  
Buffalo Milk ◽  
Animal Origin ◽  
Duplex Pcr ◽  
Mozzarella Cheese ◽  
Chain Reaction ◽  
Polymerase Chain

Molecular biology techniques have been used for species identification in food of animal origin in relatively recent years. A polymerase chain reaction (PCR) based method, the multiplex PCR, was recently applied to species identification in meat and meat products. It allows co-amplification of separate regions of a single gene or specific fragments, each typical of a different animal species in a single PCR reaction, using different pairs of primers in the same reaction mix. In the present paper, the duplex-PCR technique is proposed to identify bovine and water buffalo DNA in a single PCR assay in milk and mozzarella cheese (a typical Italian cheese, originally made from pure water buffalo milk). Because of its lower cost, undeclared bovine milk is added to water buffalo milk for making different kinds of mozzarella cheese. The results of this experiment indicate the applicability of this method, which showed an absolute specificity for the two species and a high sensitivity even down to low DNA concentrations (1 pg). In bovine and water buffalo mixtures of both milk and mozzarella cheese, the minimum concentration tested was 1% of bovine in water buffalo milk and water buffalo in bovine milk. The importance of the somatic cell content in raw milk is also discussed with special reference to the evaluation of mixtures (milk or cheese) of the two species.

scholarly journals Oxytetracycline induces DNA damage and epigenetic changes: a possible risk for human and animal health?

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3236 ◽  
Author(s):  
Adriana Gallo ◽  
Rosaria Landi ◽  
Valentina Rubino ◽  
Alessandro Di Cerbo ◽  
Angela Giovazzino ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Damage ◽  
Molecular Mechanisms ◽  
Mononuclear Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Animal Health ◽  
Animal Origin ◽  
Chain Reaction ◽  
Polymerase Chain

Background Oxytetracycline (OTC), which is largely employed in zootechnical and veterinary practices to ensure wellness of farmed animals, is partially absorbed within the gastrointestinal tract depositing in several tissues. Therefore, the potential OTC toxicity is relevant when considering the putative risk derived by the entry and accumulation of such drug in human and pet food chain supply. Despite scientific literature highlights several OTC-dependent toxic effects on human and animal health, the molecular mechanisms of such toxicity are still poorly understood. Methods Here, we evaluated DNA damages and epigenetic alterations by quantitative reverse transcription polymerase chain reaction, quantitative polymerase chain reaction, chromatin immuno-precipitation and Western blot analysis. Results We observed that human peripheral blood mononuclear cells (PBMCs) expressed DNA damage features (activation of ATM and p53, phosphorylation of H2AX and modifications of histone H3 methylation of lysine K4 in the chromatin) after the in vitro exposure to OTC. These changes are linked to a robust inflammatory response indicated by an increased expression of Interferon (IFN)-γ and type 1 superoxide dismutase (SOD1). Discussion Our data reveal an unexpected biological in vitro activity of OTC able to modify DNA and chromatin in cultured human PBMC. In this regard, OTC presence in foods of animal origin could represent a potential risk for both the human and animal health.

scholarly journals DETECTION OF CAMPYLOBACTER SPP. WITH REAL-TIME POLYMERASE CHAIN REACTION

2019 ◽  
pp. 3-7
Author(s):  
G. S. Skitovich ◽  
K. V. Serova ◽  
N. B. Shadrova ◽  
O. V. Pruntova
Keyword(s):  
Polymerase Chain Reaction ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Real Time ◽  
Carbon Dioxide Concentration ◽  
Ion Concentration ◽  
Animal Origin ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Polymerase Chain

Bacteria of Campylobacter genus are ones of the main zoonotic pathogens causing human and animal diseases. Campylobacter organisms are microaerophiles and, therefore, require low oxygen concentration (3–5%) and high carbon dioxide concentration (3–10%) for their growth. They use amino acids rather than carbons as a source of energy. Classical bacteriological methods for Campylobacter spp. detection are not always successful due to diffi culties in creating optimal conditions for their growth. Therewith, development and implementation of molecular methods for Campylobacter detection and identifi cation are of current importance. Assay for qualitative Campylobacter spp. detection with real-time polymerase chain reaction using CFX-96 thermocycler was optimized. Highly specifi c segment of 16S rRNA gene allowing identifi cation of 6 Campylobacter species: C. jejuni, C. coli, C. lari, C. upsaliensis, C. helveticus и C. hyointestinalis was selected as an amplifi cation target. Optimal magnesium ion concentration (2.5 мМ) and primer annealing temperature (58 °С) were determined. Eighteen reference strains of various bacteria were tested. Only tests of Campylobacter genus strains gave positive results. The method sensitivity was 40 target molecules. The said method was used for testing 76 samples of raw materials of animal origin. Campylobacter spp. genome was detected in 18 samples. Obtained results showed that the optimized variant of real-time polymerase chain reaction based on 16S rRNA gene amplifi cation was a specifi c, sensitive, rapid, reproducible and accurate method for qualitative detection of Campylobacter spp. in samples of raw animal materials.

scholarly journals Molecular Identification of Vancomycin Resistance and Virulence Genes in Foodborne Enterococci

2019 ◽  
Vol 70 (2) ◽  
pp. 1487
Author(s):  
T. E. MUS ◽  
F. CETINKAYA ◽  
R. CIBIK ◽  
G. DEGIRMENCI ◽  
F. B. DILER
Keyword(s):  
Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Epidemiological Surveillance ◽  
Animal Origin ◽  
Surveillance Systems ◽  
Chain Reaction ◽  
Vana Gene ◽  
Virulence Traits ◽  
Vancomycin Resistant ◽  
Polymerase Chain

The study was performed to determine the presence of vancomycin phenotyping genes and some virulence traits in enterococci species. For this purpose, a total of 42 enterococci including 6 vancomycin-resistant and 36 vancomycin-susceptible strains originated from meat/meat products and milk/dairy products were assessed for the vanA, vanB and vanC genes and agg, esp, gelE, ace and efaA virulence genes by using polymerase chain reaction or multiplex polymerase chain reaction. The vanA gene was found in 12% (n=5) of the strains and vanC gene in 50% (n=21). From these, three vanA- (E. faecalis, E. durans, E. casseliflavus) and two vanC-positive (E. durans) strains had a minimum inhibitory concentration of > 256 μg/ml as previously determined with the E-test. The strains expressing vancomycin susceptibility originating from ready-to-eat food were found to carry vanA (n=1) and vanC (n=5) genes. On the other hand, the vanB gene was not detected among strains. Moreover, no strain was found to harbor virulence traits studied. Our results indicated that resistant or susceptible enterococci from foods of animal origin can be a possible reservoir for resistance genes and may have a potential role for transfer of genetic elements among enterococci or to other bacteria. Furthermore, to develop epidemiological surveillance systems for foodborne antibiotic resistant pathogens as vancomycin-resistant enterococci and their genes responsible for resistance, primarily vanA, vanB, continues to be an essential issue all around the world. The present work provides data for foodborne enterococci isolates harboring vanA gene from Turkey.

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