Detection of ovine milk adulteration using taqman real-time pcr assay

Šnirc, Marek; Fekete, T.; Belej, Ľubomí­r; Židek, Radoslav; Golian, Jozef; Haščí­k, Peter; Zajác, Peter; Čapla, Jozef

doi:10.5219/782

Cited by 4 publications

(3 citation statements)

References 27 publications

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“…Po separácii a zafarbení sa vykonáva denzitometrické vyhodnotenie elektroforetických pásov pomocou softvéru (Sienkiewicz, Dogan a Kroh, 2006). Na detekciu falšovania ovčieho syra sú k dispozícii aj iné metódy (Zeleňáková et al, 2016;Fekete et al, 2017;Šnirc et al, 2017). a možno ich použiť v kombinácii s touto metódou na podporu platnosti výsledkov.…”

Section: Materiál a Metodikaunclassified

Food safety and quality. Scientific proceedings

2023

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We tested 68 strains of Staphylococci, (S. aureus -29, S. epidermidis -34, S. haemolyticus -23, S. warneri -16, and S. chromogenes -6). They have been isolated from various types of mastitis. Staphylococci in milk samples were isolated from acute form (27,9 %), subacute form (10,3 %), subclinical form (25 %) and latent form (36,8 %). The most common form of mastitis in dairy cows was latent mastitis. Formation of biofilm was evaluated by results of two methods: quantitative growth on CongoRed agar (CRA) and semiquantitative growth on microtiter plates (MTP). At the significance level of α = 0.05, the dependence of biofilm formation in bacteria positive using both phenotypic methods in S. aureus, S. epidermidis and S. haemolyticus was confirmed. The positivity in S. warneri and S. chromogenes determined by both methods was random. Based on our results and found dependence in comparison both methods of biofilm detection, we can recommend the simultaneous use of these methods in ideal conditions. It should be supplemented by determining the presence of genes encoding biofilm production. Keywords production of biofil:, staphylococci m, cow´s mastitis, milk ÚVOD Tvorba biofilmu ako významný faktor virulencie pomáha baktériám vyhnúť sa imunitnej obrane hostiteľa a antimikrobiálnej terapii. V chovoch prežúvavcov má biofilm vplyv na intenzitu a pretrvávanie mastitíd spôsobených v prevažnej miere baktériami z rodu Staphylococcus spp., ktoré sú najčastejšou príčinou mastitíd u prežúvavcov (Vasiľ et al., 2017).Vzhľadom na úlohu biofilmu pri perzistencii a kolonizácii stafylokokov v epiteli mliečnej žľazy, zníženie imunologickej odpovede organizmu, rezistencie na antibiotickú liečbu a ťažkosti pri eradikácii patogénov má podrobné preskúmanie a pochopenie mechanizmu tvorby biofilmu zásadný význam pri terapii príp. prevencii zápalov mliečnej žľazy. Je všeobecne známe, že baktérie produkujúce biofilm je ťažké z mliečnej žľazy odstrániť, sú často rezistentné na antibiotickú liečbu, ktorá sa na liečbu mastitíd bežne používa (Farkašová et al., 2020).Cieľom našej práce bolo stanovenie biofilmu u stafylokokov izolovaných z mlieka mastitídnych dojníc počas jedného roka. MATERIÁL A METODIKAVyšetrované vzorky mlieka sme odobrali v priebehu jedného roka od dojníc na PD Kluknava na východe Slovenska. Odbery sme uskutočnili u dojníc v štvrťročných intervaloch (marec, jún, september december). Celkom bolo vyšetrených 120 kusov dobytka. Mlieko bolo hneď po odbere vyšetrené maštaľným NK testom na detekciu typu mastitídy (akútna, subakútna, subklinická, latentná), a následne bakteriologickou kultiváciou na 5 % krvnom agare a na špeciálnych selektívnych pôdach. Následne boli stafylokokové kmene identifikované pomocou dvoch rôznych identifikačných metód: na základe biochemických enzymatických vlastností baktérií STAPHYtest 24 s identifikačným programom TNW 7.0 (Erba-Lachema, Brno, Česká republika) s presnosťou detekcie viac ako 90,0 % a na základe stanovenia

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Section: Materiál a Metodikaunclassified

Food safety and quality. Scientific proceedings

2023

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“…The presence of modified γ2-casein (with pI = 6.7), which derives from the interaction between γ2-casein and aldehydes coming from microbial metabolism, could be present on the gel . [28] There are several methods available to detect adulteration in ewe's cheese, [29][30][31] and these can be used in combination with this method to validate the results.…”

Section: 000mentioning

confidence: 99%

Detection of adulteration of traditional Slovak bryndza ewe’s cheese with cow’s lump cheese by isoelectric focusing of gamma caseins

Zajác

Beňová

Židek

et al. 2021

International Journal of Food Properties

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Slovak bryndza" is a traditional Slovak cheese manufactured by milling a lump of matured ewe's cheese or by milling a mixture of lump ewe's cheese and lump cow's cheese. The percentage of lump ewe's cheese is greater than 50% w/w dry matter. Differences in the purchase prices of ewe's and cow's milk create the potential for adulteration. The detection of cow's milk compounds by isoelectric focusing of γ-caseins after plasmolysis is a suitable method for the determination of adulteration of this traditional cheese. The analysis and final results may be affected by factors affecting laboratory activity and biological factors. Biological factors are manifested primarily as variations in the casein content of ewe's and cow's milk. Despite this variation, this method may also be used for quantification purposes. We were able to establish a reliable calibration line from the internal reference materials prepared in our laboratory. The mean composition of Slovak bryndza cheese determined in our research was protein 18.7 ± 1.9 g 100 g −1 , fat 24.0 ± 1.8 g 100 g −1 , lactose 1.5 ± 1.9 g 100 g −1 , ash 4.5 ± 0.3 g 100 g −1 , fat in dry matter 49.3 ± 2.6 g 100 g −1 , casein 18.4 ± 1.9 g 100 g −1 , gamma casein 0.60 ± 0.19 g 100 g −1, and bovine-to-ovine gamma casein ratio 0.65 ± 0.12 g 100 g −1 .

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“…Efforts have been made to develop low-cost and user-friendly PCR devices with accuracy and stability comparable to commercial alternatives [278]. Commercial PCR-based assays designed for the detection and quantitative authentication of animal species in a specific dairy product are also available in the market [279,280].…”

Section: Milk or Dairy Products Authenticity Issue Analytical Techniqmentioning

confidence: 99%

Fraud in Animal Origin Food Products: Advances in Emerging Spectroscopic Detection Methods over the Past Five Years

Hassoun

Måge

Schmidt

et al. 2020

Foods

103

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Animal origin food products, including fish and seafood, meat and poultry, milk and dairy foods, and other related products play significant roles in human nutrition. However, fraud in this food sector frequently occurs, leading to negative economic impacts on consumers and potential risks to public health and the environment. Therefore, the development of analytical techniques that can rapidly detect fraud and verify the authenticity of such products is of paramount importance. Traditionally, a wide variety of targeted approaches, such as chemical, chromatographic, molecular, and protein-based techniques, among others, have been frequently used to identify animal species, production methods, provenance, and processing of food products. Although these conventional methods are accurate and reliable, they are destructive, time-consuming, and can only be employed at the laboratory scale. On the contrary, alternative methods based mainly on spectroscopy have emerged in recent years as invaluable tools to overcome most of the limitations associated with traditional measurements. The number of scientific studies reporting on various authenticity issues investigated by vibrational spectroscopy, nuclear magnetic resonance, and fluorescence spectroscopy has increased substantially over the past few years, indicating the tremendous potential of these techniques in the fight against food fraud. It is the aim of the present manuscript to review the state-of-the-art research advances since 2015 regarding the use of analytical methods applied to detect fraud in food products of animal origin, with particular attention paid to spectroscopic measurements coupled with chemometric analysis. The opportunities and challenges surrounding the use of spectroscopic techniques and possible future directions will also be discussed.

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Detection of ovine milk adulteration using taqman real-time pcr assay

Cited by 4 publications

References 27 publications

Food safety and quality. Scientific proceedings

Food safety and quality. Scientific proceedings

Detection of adulteration of traditional Slovak bryndza ewe’s cheese with cow’s lump cheese by isoelectric focusing of gamma caseins

Fraud in Animal Origin Food Products: Advances in Emerging Spectroscopic Detection Methods over the Past Five Years

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