Hostname: page-component-586b7cd67f-rdxmf Total loading time: 0 Render date: 2024-11-20T11:41:18.896Z Has data issue: false hasContentIssue false
  • English
  • Français

Measurement of thyroid hormones in donkey (Equus asinus) blood and milk: validation of ELISA kits and evaluation of sample collection, handling and storage

Published online by Cambridge University Press:  08 September 2010

Luca Todini ,
Alessandro Malfatti ,
Elisabetta Salimei  and
Francesco Fantuz
Luca Todini*
Affiliation:
Dipartimento di Scienze Ambientali, Università di Camerino, Italy
Alessandro Malfatti
Affiliation:
Dipartimento di Scienze Ambientali, Università di Camerino, Italy
Elisabetta Salimei
Affiliation:
Dipartimento di Scienze e Tecnologie Agro-alimentari Ambientali e Microbiologiche, Università del Molise (CB), Italy
Francesco Fantuz
Affiliation:
Dipartimento di Scienze Ambientali, Università di Camerino, Italy
*
*For correspondence; e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Donkey's milk is well tolerated by human infants with cow's milk allergy and is useful in the treatment of human immune-related diseases and in the prevention of atherosclerosis. Thyroid hormones (TH) stimulate lactation and active triiodothyronine (T3) in colostrum and milk could take paracrine action supporting lactogenesis in the mother, and play physiological roles for the suckling offspring (systemic or within the gastrointestinal tract). The aims were to measure TH concentrations in donkey blood and milk, validate ELISA methods, evaluate the effects of sample collection and post-collection handling and the stability of TH in milk and blood serum and plasma samples. In milk and blood samples obtained from lactating jennies total concentrations of TH were assayed using competitive-type ELISA kits. Good validation results were obtained for both TH concentrations in blood serum and plasma and T3 in milk samples extracted with cold (−20°C) ethanol alkalinized (pH 9·0) with NH4OH. In most of the milk extract samples, thyroxine (T4) concentrations resulted below the sensitivity threshold. Intra- and inter-assay coefficients of variations of TH concentrations in different blood and milk samples were below 10%. Parallelism tests gave displacement lines parallel to those of the calibrators for both TH in blood serum and plasma and for T3 in milk extracts. Mean recovery rates were between 95% and 123%, but the concentration values approaching the highest calibrators were overestimated. Therefore, serum and plasma samples for T3 assay must be previously diluted with buffer. Both TH concentrations in blood serum and plasma and T3 in milk did not change during storage for up to 6 months at −20°C. In conclusion, the ELISA methods tested in the present study are suitable for determination of both TH concentrations in donkey blood samples, and for T3 measurement in milk, after extraction with cold alkaline ethanol.

Keywords

donkeymilk and blood thyroid hormonesELISA validationfunctional food
Type
Research Article
Information
Journal of Dairy Research , Volume 77 , Issue 4 , November 2010 , pp. 419 - 424
Copyright
Copyright © Proprietors of Journal of Dairy Research 2010

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Akasha, MA & Anderson, RR 1984 Thyroxine and triiodothyronine in milk of cows, goats, sheep and guinea pig. Proceedings of the Society for Experimental Biology and Medicine 177 360–371CrossRefGoogle Scholar
Allen, AL, Scott, WM, Cook, SJ, Fretz, PB & Doige, CE 1997 Effects of delayed serum separation and long-term storage on the measurement of thyroid hormones in equine blood samples. Veterinary Clinical Pathology 26 10–12CrossRefGoogle ScholarPubMed
Behrend, EN, Kemppainen, RJ & Young, DW 1998 Effect of storage conditions on cortisol, total thyroxine, and free thyroxine concentrations in serum and plasma of dogs. Journal of the American Veterinary Medical Association 212 1564–1568CrossRefGoogle ScholarPubMed
Bugalia, NS, Sharma, S, Garg, SL, Kumar, V & Chander, S 2000 Plasma thyroid hormones concentrations in donkey stallions. Indian Journal of Animal Science 70 471–472Google Scholar
Fazio, E, Medica, P, Cravana, C, Messineo, C & Ferlazzo, A 2007 Total and free iodothyronine levels of growing Thoroughbred foals: Effects of weaning and gender. Livestock Science 110 207–213CrossRefGoogle Scholar
Giosuè, C, Alabiso, M, Russo, G, Alicata, ML & Torrisi, C 2008 Jennet milk production during the lactation in a sicilian farming system. Animal 2 1491–1495CrossRefGoogle Scholar
GraphPad Software 2003. San Diego California USA, www.graphpad.comGoogle Scholar
Guo, HY, Pang, K, Zhang, XY, Zhao, L, Chen, SW, Dong, ML & Ren, FZ 2007 Composition, physiochemical properties, nitrogen fraction distribution, and amino acid profile of donkey milk. Journal of Dairy Science 90 1635–1643CrossRefGoogle ScholarPubMed
Monti, G, Bertino, E, Muratore, MC, Coscia, A, Cresi, F, Silvestro, L, Fabris, C, Fortunato, D, Giuffrida, MG & Conti, A 2007 Efficacy of donkey's milk in treating highly problematic cow's milk allergic children: an in vivo and in vitro study. Pediatric Allergy and Immunology 18 258–264CrossRefGoogle ScholarPubMed
Murray, MJ & Luba, NK 1993 Plasma gastrin and somatostatin, and serum thyroxine (T4), triiodothyronine (T3), reverse triiodothyronine (rT3) and cortisol concentrations in foals from birth to 28 days of age. Equine Veterinary Journal 25 237–239CrossRefGoogle ScholarPubMed
Salimei, E & Chiofalo, B 2006. Asses: milk yield and composition. In Nutrition and Feeding of the broodmare (Eds Miraglia, N & Martin-Rosset, W) Vol. 120, pp. 117–131. Wageningen Acad. Publ., Wageningen, NLCrossRefGoogle Scholar
Sheard, NF & Walker, WA 1988 The role of breast milk in the development of the gastrointestinal tract. Nutrition Reviews 46 1–8CrossRefGoogle ScholarPubMed
Slebodzinski, AB, Brzezinska-Slebodzinska, E, Nowak, J & Kowalska, K 1998 Triiodothyronine (T3), insulin and characteristics of 5′-monodeiodinase (5′-MD) in mare's milk from parturition to 21 days post-partum. Reproduction Nutrition Development 38 235–244Google ScholarPubMed
Slebodzinski, AB & Gawecka, H 1983 Passage of thyroid hormone into milk in rabbits. Endocrinologia Experimentalis 17 243–255Google ScholarPubMed
Slebodzinski, AB, Nowak, J, Gawecka, H & Sechman, A 1986 Thyroid hormones and insulin in milk: A comparative study. Endocrinologia Experimentalis 20 247–255Google ScholarPubMed
Slebodzinski, AB, Lipczak, W & Brzezinska-Slebodzinska, E 1995 Peroral administration of triiodothyronine (T3) affects absorption of immunolactaglobulins in calves. Reproduction Nutrition Development 35 383–387CrossRefGoogle ScholarPubMed
Slebodzinski, AB, Brzezinska-Slebodzinska, E, Styczynska, E & Szejnoga, M 1999 Presence of thyroxine deiodinase in mammary gland: possible modulation of the enzyme-deiodinating activity by somatotropin. Domestic Animal Endocrinology 17 161–169CrossRefGoogle ScholarPubMed
SPSS 2003. SPSS® 12.0 applications guide. Chicago, IL, SPSS Inc.Google Scholar
Tafaro, A, Magrone, T, Jirillo, F, Martemucci, G, D'alessandro, AC, Amati, L & Jirillo, E 2007 Immunological Properties of Donkey's Milk: Its Potential Use in the Prevention of Atherosclerosis. Current Pharmaceutical Design 13 3711–3717CrossRefGoogle ScholarPubMed
Todini, L 2007 Thyroid hormones in small ruminants: effects of endogenous, environmental and nutritional factors. Animal 1 997–1008CrossRefGoogle ScholarPubMed
Vita, D, Passalacqua, G, Di Pasquale, G, Caminiti, L, Crisafulli, G, Rulli, I & Pajno, GB 2007 Ass's milk in children with atopic dermatitis and cow's milk allergy: crossover comparison with goat milk. Pediatric Allergy and Immunology 18 594–598CrossRefGoogle Scholar