journal_list | How to participate | E-utilities
Cha, Chae, Park, Yoo, Son, Kim, and Lee: Effects of reduced glutathione on stress and inflammatory response in Korean native calves vaccinated with foot-and-mouth disease vaccine

Abstract

This study evaluated the effect of reduced glutathione (GSH) for the reduction of stress and inflammatory response in calves inoculated with foot-and-mouth disease (FMD) vaccine. Twenty-five calves were divided into five groups of 5 calves. The negative control (NC) did not receive any vaccination or drug treatment. The positive control (PC), GSH-25, GSH-50 and GSH-100 were intramuscularly injected with GSH at concentrations of 0, 25, 50 and 100 mg / 10 kg body weight (BW), respectively, for 3 days after FMD vaccination. On day 3, 5 and 7 post-treatment, the serum cortisol and tumor necrosis factor-α (TNF-α) levels in GSH-50 and GSH-100 were significantly decreased compared with those in PC (p < 0.05). However, there was no significant difference in the serum cortisol and TNF-α levels between GSH-100 and NC 3 and 5 days post-treatment, and between GSH-50, GSH-100, and NC 7 days post-treatment. The results from this study suggest that treatment of 50 mg / 10 kg BW GSH for 3 days is useful for the reduction of stress and inflammatory response caused by FMD vaccination in calves.

FMD outbreaks occurred in the Republic of Korea in 2000, 2002, 2010-2011 and 2014-2015 [7]. Since the FMD outbreak of 2010-2011, a national blanket vaccination policy has been enforced for ruminants and pigs as a sustainable preventive measure. Although this program is somewhat controversial, a routine vaccination program is still a central control tool against FMD virus infection [8].
In Korea, typical vaccinations were started in September 2011 using a trivalent FMD vaccine [8]. The FMD vaccine consists of inactivated viruses in a double oil-based emulsion and includes structural proteins of FMD viruses (O1 Manisa + A Malaysia + Asia 1 Shamir serotypes) [7, 8]. The vaccination practice can stress animals due to the injection process, possible inflammatory response, or adverse effects resulting from the vaccination. Growth reduction, disturbance of pregnancy, and drop in milk production due to vaccination have been observed in some cases [10].
Immune responses of animals to FMD vaccines have been extensively investigated [6-8]. However, there are few published reports on the control of stress and inflammatory responses in cattle after FMD vaccination. Therefore, this study evaluated the reductive potential of reduced glutathione (GSH) on stress and inflammatory responses in Korean native calves after FMD vaccination.
The drug (Vital-Soot®, 1,000 mg of GSH per bottle) was obtained from Dae Han New Pharm Co. Ltd. (Seoul, Korea). The inactivated trivalent FMD vaccine was purchased from Komipharm International Co., Ltd. (PRO-VAC® FMD, Korea). This study was conducted at the livestock farm of Gyeongsang National University (Jinju, Korea). Twenty-five 9-month-old Korean native calves without any history of FMD vaccination were selected and randomly divided into five groups (5 calves per group). The negative control (NC) was a non-FMD vaccination and non-drug treatment group. The positive control (PC), GSH-25, GSH-50 and GSH-100 were intramuscularly injected with GSH at levels of 0.0, 25, 50 and 100 mg / 10 kg body weight (BW), respectively, for 3 consecutive days after FMD vaccination. All animal experiments were conducted under ethics approval from the Gyeongsang National University Animal Ethics Committee in accordance with the guidelines of the Korean Council on Animal Care (GNU-180112-A0003). Blood samples were collected into a heparinized vacutainer tube from the tail vein of calves at 0, 1, 3, 5 and 7 days post-drug treatment. All blood samples were centrifuged at 2,000 × g for 10 min to separate the serum. According to the manual procedures, cortisol and TNF-α concentration in the serum were analyzed using a bovine cortisol ELISA kit and TNF-α ELISA kit (Cusabio Biotech Co. Ltd., USA), respectively. All results were represented as the mean ± SD. Statistical analyses were performed using SPSS 16.0 (SPSS Inc., Chicago, IL, USA). Data were analyzed using one-way analysis of variance (ANOVA) and the value of p < 0.05 was used as the criterion for statistical significance.
On the 1st day post-drug treatment, cortisol levels in GSH-100 were significantly decreased compared with those in PC (p < 0.05). However, cortisol levels in GSH-100 and NC were not significantly different. On the 3rd and 5th day post-treatment, cortisol concentrations in GSH-50 and GSH-100 were significantly decreased compared with those in PC (p < 0.05), but there was no significant difference between GSH-100 and NC. On the 7th day post-treatment, cortisol concentrations in GSH-50 and GSH-100 were significantly decreased compared with those in PC (p < 0.05), but there was no significant difference between GSH-50, GSH-100, and NC (Fig. 1).
Fig. 1
Change in cortisol concentrations in serum. NC, negative control (non-treated); PC, positive control. PC, GSH-25, GSH-50 and GSH-100 were intramuscularly injected with reduced glutathione at concentrations of 0, 25, 50 and 100 mg / 10 kg body weight, respectively, for 3 days post- vaccination. Means within the same day with different superscripts are significantly different (p < 0.05).
JPVM_42_037_fig_1.tif
On the 3rd and 5th day post-treatment, serum TNF-α concentrations in GSH-50 and GSH-100 were significantly decreased compared with those in PC (p < 0.05), but there was no significant difference in serum TNF-α levels between GSH-100 and NC. On the 7th day post-treatment, serum TNF-α levels in GSH-50 and GSH-100 were significantly decreased compared with those in PC (p < 0.05), and there was no significant difference in serum TNF-α levels between GSH-50, GSH-100, and NC (Fig. 2).
Fig. 2
Change in tumor necrosis factor-α (TNF-α) concentrations in serum. NC, negative control (non-treated); PC, positive control. PC, GSH-25, GSH-50 and GSH-100 were intramuscularly injected with reduced glutathione at concentrations of 0, 25, 50 and 100 mg / 10 kg body weight, respectively, for 3 days post-vaccination. Means within the same day with different superscripts are significantly different (p < 0.05).
JPVM_42_037_fig_2.tif
GSH is a powerful antioxidant due to its superior free radical scavenging capability and is capable of preventing damage to important cellular components caused by reactive oxygen species [5]. According to the results from a survey on animal responses after FMD vaccination, stress, fever and pain were observed in the vaccinated animals [8]. In cattle, many conditions, such as vaccination, weaning and transportation, are considered to be stressors. Such stressors have been shown to lead to increased blood concentrations of cortisol and activated free radical oxidation [4].
The drug (GSH-50 and GSH-100) in the present study demonstrated a potential effect on the reduction of FMD vaccine-stress in cattle 3 days post-treatment, as GSH is capable of preventing damage to important cellular components caused by excessive reactive oxygen species due to oxidative stress, a state of imbalance between the oxidant and antioxidant defense systems [1]. In a previous study, serum cortisol concentrations in calves injected with ketoprofen (3 mg/kg BW) post-surgical castration were significantly decreased compared with those in surgical castrated-calves (p < 0.05) [2]. In addition, another previous study showed that serum cortisol levels in dehorned calves intravenously treated with flunixin (2.2 mg/kg BW) were significantly decreased compared with those in the control group every day for 7 days post-dehorning (p < 0.05) [4]. However, dehorned calves orally treated with meloxicam and gabapentin at a dose of 2.2 mg/kg BW did not show reduced cortisol concentrations. On the other hand, serum cortisol levels in beef cattle were not a significantly different between the control and the group orally administered with meloxicam (1 kg/BW daily) for 7 days before and 7 days after vaccination against respiratory pathogens [9]. Considering the dosage, route and period of administration, GSH-50 and GSH-100 were more efficient than meloxicam and gabapentin in reducing stress, but less efficient than ketoprofen and flunixin. Results from this study suggest that GSH has an effect on the reduction of serum TNF-α concentration by inhibiting the nuclear transcription factor kappa B, which is responsible for the activation of some pro-inflammatory cytokines such as TNF-α and interleukins [11].
In a previous study, serum TNF-α concentrations in beef cattle were not significantly different between the control group and the group orally administered with meloxicam (1 mg/kg BW) for 7 days from one day before to six days after vaccination against respiratory pathogens [9]. In addition, TNF-α levels in another previous study decreased in FMD vaccinated-pigs supplemented with germanium biotite (30 kg/ton feed) for 5 weeks, but there were no significant differences between the treated pigs and the control group [6]. Considering the animal species, dosage, route and period of administration, GSH-50 and GSH-100 in this study were more effective than meloxicam and germanium biotite in terms of the decrease in serum TNF-α level. GSH has an effect on the reduction of serum TNF-α concentration by inhibiting the nuclear transcription factor kappa B, which is responsible for the activation of some pro-inflammatory cytokines such as TNF-α and interleukins [3, 11].
In the present study, the efficacy of GSH-50 and GSH-100 against stress and inflammatory response caused by FMD vaccination was demonstrated by the decrease in serum cortisol and TNF-α concentrations. Results from this study suggest that intramuscular injection of GSH at a concentration of 50 mg/10 kg BW for 3 consecutive days after FMD vaccination may be useful for the release of stress and inflammation from FMD vaccination in calves.

ACKNOWLEDGEMENTS

This research was supported by Dae Han New Pharm Co. Ltd. (Seoul, Korea).

REFERENCES

1. Akyol S, Erdogan S, Idiz N, Celik S, Kaya M, Ucar F, Dane S, Akyol O. The role of reactive oxygen species and oxidative stress in carbon monoxide toxicity:an in-depth analysis. Redox Rep. 2014; 19:180–189. DOI: 10.1179/1351000214Y.0000000094. PMID: 24773392.
[CrossRef] [Google Scholar]
2. Earley B, Crowe MA. Effects of ketoprofen alone or in combination with local anesthesia during the castration of bull calves on plasma cortisol, immunological, and inflammatory responses. J Anim Sci. 2002; 80:1044–1052. DOI: 10.2527/2002.8041044x. PMID: 12002311.
[CrossRef] [Google Scholar]
3. Ghosh S, Hayden MS. New regulators of NF-kB in inflammation. Nat Rev Immunol. 2008; 8:837–848. DOI: 10.1038/nri2423. PMID: 18927578.
[CrossRef] [Google Scholar]
4. Glynn HD, Coetzee JF, Edwards-Callaway LN, Dockweiler JC, Allen KA, Lubbers B, Jones M, Fraccaro E, Bergamasco LL, KuKanich B. The pharmacokinetics and effects of meloxicam, gabapentin, and flunixin in postweaning dairy calves following dehorning with local anesthesia. J Vet Pharmacol Ther. 2013; 36:550–561. DOI: 10.1111/jvp.12042. PMID: 23473342.
[CrossRef] [Google Scholar]
5. Korge P, Calmettes G, Weiss JN. Increased reactive oxygen species production during reductive stress:The roles of mitochondrial glutathione and thioredoxin reductases. Biochim Biophys Acta. 2015; 1847:514–525.
[CrossRef] [Google Scholar]
6. Lee JA, Jung BG, Jung M, Kim TH, Yoo HS, Lee BJ. Dietary germanium biotite supplementation enhances the induction of antibody responses to foot-and-mouth disease virus vaccine in pigs. J Vet Sci. 2014; 15:443–447. DOI: 10.4142/jvs.2014.15.3.443. PMID: 24690605. PMCID: PMC4178148.
[CrossRef] [Google Scholar]
7. Lee JH, Kang IJ, Kim AR, Noh YS, Chung HC, Park BK. Increased humoral antibody response of foot-and-mouth disease virus vaccine in growing pigs pre-treated with poly-γ-glutamic acid. J Vet Sci. 2016; 17:253–256. DOI: 10.4142/jvs.2016.17.2.253. PMID: 26645341. PMCID: PMC4921674.
[CrossRef] [Google Scholar]
8. Lee HS, Lee NH, Seo MG, Ko YJ, Kim B, Lee JB, Kim JS, Park S, Shin YK. Serological responses after vaccination of growing pigs with foot-and-mouth disease trivalent (type O, A and Asia1) vaccine. Vet Microbiol. 2013; 164:239–245. DOI: 10.1016/j.vetmic.2013.02.012. PMID: 23490554.
[CrossRef] [Google Scholar]
9. Rodrigues MC, Cooke RF, Marques RS, Arispe SA, Keisler DH, Bohnert DW. Effects of oral meloxicam administration to beef cattle receiving lipopolysaccharide administration or vaccination against respiratory pathogens. J Anim Sci. 2015; 93:5018–5027. DOI: 10.2527/jas.2015-9424. PMID: 26523594.
[CrossRef] [Google Scholar]
10. Yeruham I, Yadin H, Haymovich M, Perl S. Adverse reactions to FMD vaccine. Vet Dermatol. 2001; 12:197–201. DOI: 10.1046/j.0959-4493.2001.00221.x. PMID: 11493403.
[CrossRef] [Google Scholar]
11. Zhang H, Liu H, Zhou L, Yuen J, Forman HJ. Temporal changes in glutathione biosynthesis during the lipopolysaccharide-induced inflammatory responses of THP-1 macrophages. Free Radic Biol Med. 2017; 113:304–310. DOI: 10.1016/j.freeradbiomed.2017.10.010. PMID: 28993271.
[CrossRef] [Google Scholar]
Formats:
Article | 
PDF LinksPDF(647K) | PubReaderPubReader | EpubePub | 
Download Citation
Share  |
         
METRICS
290
View
10
Save
0
Cited-By
In This Page: