December 1, 2021


Hypothesis: Betaine decreases the pathogenicity of Covid-19.

Dietary Betaine Improves Intestinal Barrier Function and Ameliorates the Impact of Heat Stress in Multiple Vital Organs as Measured by Evans Blue Dye in Broiler Chickens

Majid Shakeri

Simple Summary

Heat stress alters the normal physiological status, compromising the function of organs such as the small intestine. However, evidence exists of a wider distribution of organ dysfunction, stemming from factors such as a reduction in blood flow due to redistribution to the skin for increased radiant heat loss to the environment. Simultaneously, assessing organ dysfunction at multiple locations presents technical difficulties, and hence studies are lacking. Therefore, the aim of this experiment was to determine the pattern of Evans Blue Dye distribution as a cost-effective indicator of organ dysfunction in HS chickens supplemented with betaine. The results showed that Evans Blue Dye concentration increased in the kidney and muscle during heat stress, while such concentration was reduced with betaine. Therefore, betaine could improve the broiler’s tolerance to heat stress, and Evans Blue Dye may be a useful tool for investigating the effects of heat stress on broiler organ dysfunction.


In a 2 × 2 factorial design, 60 male Ross-308 broilers were fed either a control or 1 g/kg betaine diet and housed under thermoneutral (TN) or heat stress (HS) conditions. Broilers were acclimated to diets for 1 week under TN (25 °C), then either kept at TN or HS, where the temperature increased 8 h/day at 33 °C and 16 h/day at 25 °C for up to 10 days. Respiration rate (RR) was measured at four time points, and on each of 1, 2, 3, 7 and 10 days of HS, 12 broilers were injected with 0.5 mg/kg of Evans Blue Dye (EBD) solution to quantify regional changes in tissue damage. Betaine was quantified in tissues, and ileal damage was assessed via morphometry and transepithelial resistance (TER). Heat stress elevated RR (p < 0.001) and resulted in reduced villous height (p = 0.009) and TER (p < 0.001), while dietary betaine lowered RR during HS (p < 0.001), increased betaine distribution into tissues, and improved ileal villous height (p < 0.001) and TER (p = 0.006). Heat stress increased EBD in the muscle and kidney of chickens fed the control diet but not in those receiving betaine. Overall, these data indicate that supplemented betaine is distributed to vital organs and the gastrointestinal tract, where it is associated with improved tolerance of HS. Furthermore, EBD markers help reveal the effects of HS on organs dysfunction.