H. Hasan ORUÇ, K. Wilson RUMBEIHA, Steve ENSLEY, Chris OLSEN, E. Dwayne SCHRUNK

Simultaneous detection of six different groups of antimicrobial drugs in porcine oral fluids using A biochip array-based immunoassay

Çalışmanın amacı Biochip array-based immunoassay yöntemi ile 6 farklı grup antibakteriyel ilacın deneysel şartlarda toplanmış domuz ağız sıvısında tespit edilip edilemediğini; temiz ve kirli domuz ağız sularında bu antimikrobiyallerin tespit edilebilirliğini ve bu metotla, rasyonlarında sürekli olarak klortetrasiklin bulunan domuzların ağız sıvılarında klortetrasiklinin tespit edilebilirliğini belirlemektir. Bu metod, aynı anda altı farklı grup antimikrobiyalin süt, idrar, et, bal ve yem numunelerinde miktarlarının belirlenmesinde kullanılmaktadır, ancak ağız sıvılarında tespit edilebilirlikleri henüz araştırılmamıştır. Yapılan farklı ekstraksiyon çalışmalarından sonra bir santrifüj metodu seçildi. Bu metodla hedef ilaç olan norfloksasin, seftifor, florfenikol, streptomisin, taylosin ve tetrasiklin temiz ve kirli domuz ağız sıvısında tespit edildi. Ancak, kirli ağız sularında tetrasiklinin geri kazanımı oldukça düşük olarak elde edildi. Klortetrasiklin çiftlikteki domuzların ağız sıvılarında 176.0-698.6 ppb arasında tespit edildi. Sonuç olarak, bu test ile kirli ağız sıvısında tetrasiklin aranması dışında, altı farklı gruptaki hedef antibakteriyel ilaç domuzların ağız sıvısında kolaylıkla tespit edilebilmektedir.

Anahtar Kelimeler:

ilaç kalıntıları, doğruluk, klortetrasiklin, diyetler, yemler, gıda hijyeni, gıda güvenirliği, bal, bağışıklık deneyi, domuz eti, domuz, teknikler, idrar

Domuz ağız sıvısında altı farklı grup antimikrobiyal ilacın aynı anda biochip array-based immunoassay ile tayini

The objectives of this study were 1) to determine whether a biochip array-based immunoassay could be used to detect 6 different group antimicrobials in porcine oral fluids collected under experimental conditions; 2) to determine the feasibility of antimicrobial detection in clean versus dirty oral fluid samples; and 3) to determine if the assay could be used to detect chlortetracycline in oral fluids of swine ingesting a routine diet containing chlortetracycline. Biochip array-based immunoassay is currently used for simultaneous detection and quantitation of different group six antimicrobials in milk, urine, meat, honey, and feed. The assay had not been investigated for potential use in oral fluids. Following evaluation of different extraction procedures, a centrifugation method was chosen. Results showed that of the six target drugs, norfloxacin, ceftiofur, florfenicol, streptomycin, tylosin and tetracycline could be detected in both clean and dirty oral fluid samples. However, tetracycline was not well recovered in dirty samples. Chlortetracycline from tetracycline group was detected in all oral fluid samples collected from the field at concentrations ranging from 176.0-698.6 ppb. In conclusion, this assay can detect all target antibacterials from different groups in clean and dirty oral fluid samples with the exception of tetracycline which was not well-recovered from in dirty samples.

Keywords:

drug residues, accuracy, chlortetracycline, diets, feeds, food hygiene, food safety, honey, immunoassay, pigmeat, pigs, techniques, urine,

PDF

___

1. De Almeida Pdel V, Gregio AM, Machado MA, de Lima AA, Azevedo LR: Saliva composition and functions: A comprehensive review. J Contemp Dent Pract, 9, 72-80, 2008.
2. Pink R, Simek J, Vondrakova J, Faber E, Michl P, Pazdera J, Indrak K: Saliva as a diagnostic medium. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub, 153 (2): 103-110, 2009.
3. Haeckel R: Factors influencing the saliva/plasma ratio of drugs. Ann Ny Acad Sci, 20 (694): 128-142, 1993.
4. Barza M: Challenges to antibiotic activity in tissue. Clin Infec Dis, 19, 910- 915, 1994.
5. Haeckel R, Hanecke P: Application of saliva for drug monitoring. An in vivo model for transmembrane transport. Eur J Clin Chem Clin Biochem, 34, 171-191,1996.
6. Randox Manual: Antimicrobial Array II (AM II), EV 3524 Manual, 2011.
7. Prickett JR, Zimmerman JJ: The development of oral fluid-based diagnostics and applications in veterinary medicine. Anim Health Res Rev, 11 (2): 207-116, 2010.
8. Prickett J, Simer R, Christopher-Hennings J, Yoon KJ, Evans RB, Zimmerman JJ: Detection of porcine reproductive and respiratory syndrome virus infection in porcine oral fluid samples: A longitudinal study under experimental conditions. J Vet Diagn Invest, 20 (2): 156-63, 2008.
9. Kittawornrat A, Prickett J, Chittick W, Wang C, Engle M, Johnson J, Patnayak D, Schwartz T, Whitney D, Olsen C, Schwartz K, Zimmerman J: Porcine reproductive and respiratory syndrome virus (PRRSV) in serum and oral fluid samples from individual boars: Will oral fluid replace serum for PRRSV surveillance? Virus Research, 154 (1-2): 170-176, 2010.
10. Meiszberg A, Karriker L, Zimmerman J, Irwin C, Coetzee J: Detection of ceftiofur and oxytetracycline in oral fluids of swine with a pen-side competitive ELISA test after intramuscular injection. J Vet Pharm Ther, 34 (5): 515-517, 2011.
11. APVMA: Australian Pesticides and Veterinary Medicines Authority Guidelines, 2004.
12. Minitab13: Minitab Inc. State College, PA, USA, 2001.
13. Kahn CM: Antibacterial agents, tetracyclines general properties. In, Kahn CM (Ed): The Merck Veterinary Manual. 10th ed., pp. 2289-2293, Merck and Co, Whitehouse Station, New Jersey, 2010.