Effect of Astaxanthin supplementation on liver profile in growing Vrindavani calves exposed to thermal stress

Introduction

The country economy is mainly depends upon agriculture and livestock sector. Livestock sector plays a vital and significant role in increasing the GDP of India. In 2050, the demand for animal products worldwide is predicted to nearly treble. In the meantime, current climate models predicted that temperatures may rise by 0.2 °C every ten years, bringing the global average surface temperature closer to 4.0 °C by 2100 (IPCC 2007). India now has 192.52 million cattle (20 th livestock census). Cattles are mostly raised in subtropical climates. Temperature, humidity, precipitation, air movement, radiation, barometric pressure, and ionisation are some of the components that make up climate (Maurya et al., 2015) [15] . Globally, different regions have different climates that are influenced by several elements such as latitude, dominant winds, evaporative conditions, water supply, elevation, and proximity to mountains (West, 2003) [21] . It is believed that the temperature ranges for lactating dairy cows is between -0.5 and 20 °C in temperature region of the world (West, 2003) [21] . Livestock production is expected to be negatively impacted by climate change. Climate change poses a serious threat to livestock production because of its impact on diseases, breeding, biodiversity, water availability, feed and forage quality and milk Production (Godde et al., 2021) [9] . The harsh climate badly affects the liver function which depicts in the change in various enzymes. The ALT and AST are two of the most reliable markers of liver condition (An-Qiang et al., 2009) [4] . Blood ALT and AST values are greater in cross-bred dairy calves exposed to heat stress (Berian et al., 2019) [5] . In growing and adult cattle, summer and the hot, humid season showed considerably greater plasma enzyme activity (ALT, AST) than winter and spring (Bhan et al., 2013) [6] . Astaxanthin, an oxygenated carotenoid, is included in the class of carotenoids known as xanthophylls, which is mostly composed of lutein, zeaxanthin, β-cryptoxanthin, and canthaxanthin (Majumder et al., 2024) [14] . The primary colourful pigment found in seafood, such as salmon, trout, red sea bream, prawns, lobster, etc., is called astaxanthin. Astaxanthin contains keto groups, hydroxyl groups, and conjugated double bonds. It has characteristics that are both lipophilic and hydrophilic (Majumder et al., 2024) [14] . Astaxanthin forms an internal and exterior link with the cell membrane; it exhibits higher biological activity than other antioxidants (Ambati et al., 2014) [3] . Astaxanthin molecules include both hydroxyl and ketone groups, which make them more polar and improve membrane function, the ionone rings at both ends of the molecule have a functional role in scavenging the free radicals (Majumder et al., 2024) [14] . The major source of natural Astaxanthin is Haematococcus pluvialis and most of studies it has been extensively used (Higuera-Ciapara et al., 2006) [10] . Numerous research demonstrated the beneficial effects of astaxanthin on immunity, antiinflammatory, photoprotective, and ocular health, among other critical applications in the nutraceuticals, cosmetics, and food and feed sectors (Soren et al.,2017) [20] . The antioxidant activity of astaxanthin is 100 times more than that of α-tocopherol and up to 10 times higher than that of other carotenoids such as zeaxanthin, lutein, canthaxantin, and β-carotene. As a result, astaxanthin is referred regarded as "super vitamin E." (Somagond et al.,2022) [19] . In this study dietary Astaxanthin has been used as an antioxidant to ameliorate heat stress.

Materials and Methods

The study was carried out at the ICAR-Indian Veterinary Research Institute in Bareilly (UP) at the Nuclear Research Laboratory and the Physiology and Climatology Division. The proposed experiment was conducted in a psychrometric chamber by the Physiology and Climatology Division of the Indian Veterinary Research Institute, Izatnagar Bareilly (UP). A total of thirty crossbred calves bearing good health, aged between one and one and a half years, were used in the experiment. Animals were divided into 5 groups. Group I was non heat stress control group (NHSC). Group II was Heat stress group, where animals were exposed to heat stress (HS). Group III was heat stress with Astaxanthin supplemented group (HS+A). Group IV was Astaxanthin supplemented group and animals were kept in normal shed without any heat stress. Group V where animal kept under direct sunlight with Astaxanthin supplementation. Dietary supplementation of Astaxanthin was given at 150mg/day/animal. The duration of all experimental groups of animal was 28 days for each group. Blood samples were collected from animal on weekly basis. Blood samples were collected on pre exposure (7 day before exposure) and 1st, 7th, 14th, 21st, 28 th day of exposure at respective schedule of experiment. In addition to above, all the animals were given 35 th day recovery period. After blood collection serum sample were separated for the estimation of ALT and AST. The data collected throughout the experimental period was analyzed by using two way ANOVA.

Results and Discussion

The results of different parameters of Liver profile test i.e ALT and AST are being presented in the

Alanine aminotransferase (ALT)

The Mean ±SE value of control and treatment (HS, HS+A, Sh+A, and En+A) group have been presented in the Table 1 and Figure 1. Prior to exposure, there was no significant difference in the ALT levels between the control, HS, HS+A, En+A, and Sh+A, respectively. On the first day of the trial, there was a significant (p<0.05) rise in the ALT values in the HS, HS+A, and En+A groups of animals as compared to the NHSC and Sh+A groups. The 7th, 14th, 21st, and 28th days of the experiment showed the same pattern in the variation of ALT levels for the various treatments (HS, HS+A, and En+A) respecctively. After 35 days of recovery period, there was no significant trend in the variation in ALT for any of the treatments. There was no significant difference on Pre exposure, 1st, 7th, 14th, 21st, 28th and 35th day in control and Sh+A group of animal. As study progress there was significant increase (P<0.05) in ALT level on 7th, 14th, 21st day of study period compare to pre exposure and recovery period. Yadav et al. (2016) [22] reported that the cattle kept in a climate chamber showed greater serum ALT activity. As temperature rises, serum ALT activity rises as observed by Sharma et al. (2011) [18] . According to Cerutti et al. (2018) [8] , calves' serum ALT activity was higher in the summer than in the winter. Kittur et al. (2024) [13] found that goats experiencing heat stress have elevated serum ALT levels but AST levels did not increased. Astaxanthin supplementation reduces the impact of heat stress in goats (Kittur et al., 2024) [13] . All Previous study results support our finding, where we found that heat stress increase in serum ALT level and supplementation Astaxanthin causes an ameliorative effect in growing Vrindavani cattle by reduction in ALT level. Before the exposure, there was no significant (p<0.05) difference in the AST values amongst control, HS, HS+A, En+A, and Sh+A, respectively. On the first day of the trial, there was a notable (p<0.05) rise in the AST values in the HS, HS+A, and En+A groups of animals as compared to the NHSC and Sh+A groups. The 7th, 14th, 21st, and 28th days of study for HS, HS+A, and En+A showed the similar pattern in the change of AST levels. After 35 days of the recovery period, there was no discernible pattern in the variation in AST level for any of the treatments; it was essentially unchanged from the pre-exposure phase.

Following a statistical analysis of the data, variations in AST were found between the pre-exposure, 1, 7, 14, 21, 28 and 35 days. Over the course of the investigation, it was observed that there was no significant difference (P>0.05) in the animals in the control group and Sh+A group. AST was found to be significantly (p<0.05) greater in the HS group of animal throughout the duration of study. As the investigation went on, the AST increased significantly (P<0.05) on days 7th, 14th, 21st as compared to preexposure and 35 days of recovery period. The highest AST value was seen on the 7th, 14th, and 21st days of the trial.

On the 28th day of the study, the AST value was lower but remained significantly (P<0.05) greater than the preexposure and recovery period. The AST concentration for the two treatments i.e. HS+A and En+A followed same trend like HS group of animals. Bueno et al. (2017) [7] found elevated serum AST activity in broiler after their heat exposure. High summertime ambient temperatures may raise transaminase enzyme activity and have negative effects on thyroid, liver, and kidney function as reported by Ramadan et al. (2015) [16] . Rathwa et al.

(2017) [17] , also observed higher levels of AST and ALT in the summer than in the winter. Aleena et al. (2020) [2] worked on heat-stressed Osmanabadi, Malabari, and Salem Black goats and reported that the Osmanabadi had significantly higher levels of AST than the Malabari, and Salem Black goat respectively. In accordance to our research finding on Astaxanthin supplementation in heat cattle Islam et al. (2017) [12] also reported a reduced in AST and ALT level in rat treated with Astaxanthin.

Conclusion

The HS group had the highest ALT and AST values. The result showed that there is significant difference between NHSC group and heat exposed group HS, HS+A and En+A group. There was no significant (P>0.05) difference between NHSC and Sh+A group. On the fourteenth day of the research period, highest value of AST and ALT value was observed in all heat exposed and direct sunlight exposed group. The ALT and AST levels were significantly lower in the Sh+A group (as compared to HS+A and En+A) among the supplemented groups. The finding of our study showed a negative effect of heat stress on liver profile of animal by increasing the ALT and AST level in the blood serum. Astaxanthin supplementation normalize the ALT and AST level in the serum which is an indicative of ameliorative effect of Astaxanthin in heat stressed growing Vrindavani Cattle.

References

1. Aleena J, Sejian V, Krishnan G, Bagath M, Pragna P, Bhatta R. Heat stress impact on blood biochemical response and plasma aldosterone level in three different indigenous goat breeds. J Anim Behav. 2020;8(4):26675.
2. Ambati RR, Phang SM, Ravi S, Aswathanarayana RG. Astaxanthin: Sources, extraction, stability, biological activities and its commercial applications -A review. Mar Drugs. 2014;12(1):128-52. DOI:10.3390/md12010128.
3. An-Qiang L, Zhi-Sheng W, An-Guo Z. Effect of chromium picolinate supplementation on early lactation performance, rectal temperatures, respiration rates and plasma biochemical response of Holstein cows under heat stress. Pak J Nutr. 2009;8(7):940-5.
4. Berian S, Gupta SK, Sharma S, Ganai I, Dua S, Sharma N. Effect of heat stress on physiological and hematobiochemical profile of cross bred dairy cattle. Indian J Anim Res. 2019;9(1):95-101.
5. Bhan C, Singh SV, Hooda OK, Upadhyay RC, Beenam MV, Mangesh V. Influence of temperature variability on physiological, hematological and biochemical profile of growing and adult Sahiwal cattle. J Environ Res Develop. 2012;7(2):986-994.
6. Bueno JPR, de Mattos Nascimento MRB, da Silva Martins JM, Marchini CFP, Gotardo LRM, de Sousa GMR. Effect of age and cyclical heat stress on the serum biochemical profile of broiler chickens. Semina: Ciências Agrárias. 2017;38(3):1383-1392.
7. Cerutti RD, Scaglione MC, Arfuso F, Rizzo M, Piccione G. Seasonal variations of some hematochemical parameters in Holstein bovine under the same livestock conditions. Veterinarski Arhiv. 2018;88(3):309-321.
8. Godde CM, Mason-D'Croz D, Mayberry DE, Thornton PK, Herrero M. Impacts of climate change on the livestock food supply chain: A review of the evidence. Glob Food Secur. 2021;28:100488.
9. Higuera-Ciapara I, Felix-Valenzuela L, Goycoolea FM. Astaxanthin: A review of its chemistry and applications. Crit Rev Food Sci Nutr. 2006;46(2):185196.
10. Islam MA, Al Mamun MA, Faruk M, Islam MTU, Rahman MM, Alam MN. Astaxanthin ameliorates hepatic damage and oxidative stress in carbon tetrachloride-administered rats. Pharmacognosy Res. 2017;9(Suppl 1):84.
11. Kittur PM, Karthiga K, Sarma L, Gururaj AV, Lata K, Verma V, Samad HA, Chouhan VS, Singh G, Maurya VP. Astaxanthin supplementation ameliorates simulated heat stress by regulating physio-biochemical responses in Sirohi goats. Vet Res Commun. 2024;1-8.
12. Majumder S, Kant L, Kittur PM, Das P, Yadav P, Samad HA. Astaxanthin: A promising antioxidant to ameliorate heat stress and improve production potential in animals. J Exp Zool India. 2024, 27(1).
13. Ramadan TA, Sharma RK, Phulia SK, Balhara AK, Ghuman SS, Singh I. Effects of melatonin and controlled internal drug release device treatment on blood metabolites of buffalo heifers during out-ofbreeding season under tropical conditions. Egyptian J Anim Prod. 2015;52(4):9-17.
14. Rathwa SD, Vasava AA, Pathan MM, Madhira SP, Patel YG, Pande AM. Effect of season on physiological, biochemical, hormonal, and oxidative stress parameters of indigenous sheep. Vet World. 2017;10(6):650.
15. Sharma AK, Kataria N. Effect of extreme hot climate on liver and serum enzymes in Marwari goats. Indian J Anim Sci. 2011;81(3):293.
16. Somagond YM, Singh SV, Deshpande A. Effect of dietary supplementation of astaxanthin, prill fat and combination on stress indicators, milk yield and composition during heat stress in buffaloes. Biol Rhythm Res. 2022;53(5):665-675.
17. Soren S, Singh SV, Kumar S. Effect of Astaxanthin supplementation on semen (Karan Fries bulls) storage at 5°C. Int J Curr Microbiol Appl Sci. 2017;6(5):23-28.
18. West JW. Effects of heat stress on production in dairy cattle. J Dairy Sci. 2003;86(6):2131-2144.
19. Yadav B, Singh G, Wankar A, Dutta N, Chaturvedi VB, Verma MR. Effect of simulated heat stress on digestibility, methane emission and metabolic adaptability in crossbred cattle. Asian-Australas J Anim Sci. 2016;29(11):1585.