Effects of Bedding Change Frequency on Lipid Peroxidation, Antioxidant Status, and Histopathological Alterations in Rats

Abstract

The aim of this research was to evaluate the effect of bedding change frequency on oxidative stress parameters of rats. In the experiment; animals were randomly and equally divided into 4 groups, each group consisted of 6 male rats. Group 1; bedding change every day, Group 2; bedding change once in 2 days, Group 3; bedding change once in 4 days, and Group 4; bedding change once in a week. Dust-free wood shavings were used as bedding material and the study lasted for 2 months. At the end of experimental period, MDA levels were found to be increased in blood, liver, kidney, heart, brain, and lung of groups with longer intervals of bedding change whereas GSH levels of these tissues were decreased. It was also found that SOD and CAT activities were higher in erythrocyte and lung tissues in Group 1 than the other groups. Moreover, notable histopathological alterations were observed in the tissues of longer intervals of bedding change (especially, group 3 and 4). As a result, it has been determined that long periods of bedding change in animals causes oxidative stress, tissues damages, and these alterations adversely affect the life quality of laboratory animals.

Keywords

• bedding
• rat
• oxidative stress

Sıçanlarda Altlık Değişim Sıklığının Lipid Peroksidasyon, Antioksidan Durum ve Histopatolojik Değişiklikler Üzerine Etkileri

Öz

Bu araştırmanın amacı sıçanlarda altlık değişim sıklığının oksidatif stres parametreleri üzerine etkisi değerlendirmektir. Çalışmada hayvanlar her grupta 6 erkek sıçan olacak şekilde rastgele ve eşit olarak 4 gruba ayrıldı. Gruplar: Grup 1; her gün altlığı değişen, Grup 2; 2 günde bir altlığı değişen, Grup 3; 4 günde bir altlığı değişen ve Grup 4; haftada bir altlığı değişen şeklinde oluşturuldu. Çalışma 2 ay olacak şekilde planlandı ve altlık malzemesi olarak tozsuz ağaç talaşı kullanıldı. Çalışmanın sonunda, daha uzun aralıklarla altlık değişimi olan gruplarda kan, karaciğer, böbrek, kalp, beyin ve akciğer dokularında MDA düzeylerinin arttığı, bu dokulardaki GSH düzeylerinin ise azaldığı görülmüştür. Ayrıca, eritrosit ve akciğer dokularında SOD ve CAT aktivitelerinin Grup 1'de, diğer gruplardan daha yüksek olduğu belirlendi. Ayrıca, daha uzun aralıklarla altlıkları değişen grupların (özellikle grup 3 ve 4) beyin, akciğer, kalp, karaciğer ve böbrek dokularında histopatolojik değişiklikler gözlendi. Sonuç olarak, hayvanlarda uzun aralıklarla altlık değişiminin oksidatif strese, doku hasarlarına yol açarak laboratuvar hayvanlarının yaşam kalitesini olumsuz etkilediği belirlenmiştir.

Anahtar Kelimeler

• altlık
• sıçan
• oksidatif stres

References

1. Aebi H. Catalase in vitro. In: Bergmeyer, U. (Ed.), Methods of Enzymatic Analysis.Academic Press, New York and London, 1974; pp. 673–677.
2. Beutler E, Duron O, Kelly BM, Improved method for the determination of blood glutathione. J Lab Clin Med. 1963; 61: 882–888.
3. Burn CC, Peters A, Day MJ, Mason GJ. Long-term effects of cage-cleaning frequency and bedding type on laboratory rat health, welfare, and handleability: a cross-laboratory study. Lab anim. 2006; 40(4): 353-370.
4. Castelhano-Carlos MJ, Baumans V. The impact of light, noise, cage cleaning and in-house transport on welfare and stress of laboratory rats. Laboratory animals, 2009; 43(4): 311-327.
5. Dean SW. Environmental enrichment of laboratory animals used in regulatory toxicology studies. Lab Anim. 1999; 33(4): 309–327.
6. Domer DA, Erickson RL, Petty, JM, Bergdall VK, Hickman-Davis JM. Processing and treatment of corncob bedding affects cage-change frequency for C57BL/6 mice. J Am Assoc Lab Anim Sci. 2012; 51(2): 162-169.
7. Drabkin DL, Austin JH. Spectrophotometric studies. II. Preparations from washed blood cells; nitric oxide hemoglobin and sulfhemoglobin. J Biol Chem. 1935; 112:51–65.
8. Draper HH, Hardley M. Malondialdehyde determination as index of lipid peroxidation. Methods Enzymol. 1990; 186: 421–431.

9. Durackova Z. Some current insights into oxidative stress. Physiol Res. 2010; 59: 459–469.
10. Ferrecchia CE, Jensen K, Van Andel R. Intracage ammonia levels in static and individually ventilated cages housing C57BL/6 mice on 4 bedding substrates. JAALAS. 2014; 53(2): 146-151.
11. Horn MJ, Hudson SV, Bostrom LA, Cooper DM. Effects of cage density, sanitation frequency, and bedding type on animal wellbeing and health and cage environment in mice and rats. JAALAS. 2012; 51(6): 781-788.
12. Kucukkurt I, Ince S, Fidan AF, Ozdemir A. The effects of dietary supplementation of different amount of Yucca schidigera powder (Sarsaponin 30 ®) on blood and tissue antioxidant defense systems and lipid peroxidation in rats. J Anim Vet Adv. 2008; 7: 1413–1417.
13. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193: 265–275.
14. Luck H. Catalase. In: Bergmeyer, H.U. (Ed.), Methods in Analysis. Academy Press, London, 1955.
15. Mason GJ, Burn C, Peters A. Acute effects of cage cleaning at different frequencies on laboratory rat behaviour and welfare. Anim Welf. 2006; 15: 161-171.
16. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979; 95: 351–358.
17. Potgieter FJ, Wilke PI. Laboratory animal bedding and nesting materials for rodents and lagomorphs. J S Afr Vet Assoc. 1993; 64: 144–148.
18. Sisein EA. Biochemistry of free radicals and antioxidants. SAJB. 2014; 2: 110–118.
19. Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem. 1988; 34: 497–500.
20. Valko M, Rhodes C, Moncol J, Izakovic MM, Mazur M. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem-biol Interact. 2006; 160(1): 1-40.
21. Winterbourn CC, Hawkins RE, Brain M, Carrell RW. The estimation of red cell superoxide activity. J Lab Clin Med. 1975; 55: 337–341.
22. Yildirim F, Yildirim BA, Yildiz A, Terim K, Kübra A, Cengiz S, Özdemir S. Evaluation of perlite, wood shavings and corncobs for bedding material in rats. J S Afr Vet Assoc. 2017; 88(1): 1-7.