Comparative evaluation of dietary supplementation with turmeric, aloe vera, moringa and shatavari for mitigation of heat stress and improvement of production parameters in lactating Murrah buffaloes

Introduction

Buffaloes contribute significantly to the dairy economy of many tropical and subtropical countries, particularly India, which possesses the largest buffalo population in the world (Darji, et al., 2024) [4] . Buffalo milk is highly valued due to its higher fat, protein, and total solids content (Chen et al., 2024) [14] . However, buffaloes are highly susceptible to heat stress because of their dark skin colour, sparse hair coat, lower density of sweat glands, and limited capacity for evaporative cooling (Jasinski et al., 2023) [17] . Under conditions of elevated ambient temperature and humidity, buffaloes experience considerable physiological stress, leading to reduced feed intake, impaired metabolic functions, compromised immune responses, and decreased milk production (Rashid et al., 2024) [16] .

International Journal of Advanced Biochemistry Research https://www.biochemjournal.com

Heat stress has emerged as a major challenge to sustainable buffalo production owing to climate change and the increasing frequency of heat waves (Sejian et al., 2018 [18] . When the Temperature-Humidity Index (THI) exceeds the comfort threshold, buffaloes exhibit increased rectal temperature, respiration rate, pulse rate, and oxidative stress (Polsky & Keyserlingk 2017) [28] . These physiological alterations disrupt endocrine and metabolic homeostasis, resulting in reduced nutrient utilization and productivity. Economic losses associated with heat stress include decreased milk yield, fat and SNF% and reduced overall farm profitability (Gujar et al., 2023) [19] . Conventional approaches to alleviate heat stress, such as cooling systems, fans, sprinklers, foggers, and wallowing facilities, are often expensive and difficult to implement for smallholder farmers (Das et al., 2016) [23] . Consequently, there is growing interest in natural and cost-effective nutritional interventions that can enhance the resilience of dairy animals to thermal stress (Wang et al., 2024) [33] .

Medicinal plants and herbal feed additives have gained considerable attention due to their antioxidant, antiinflammatory, adaptogenic, immunomodulatory, and metabolic regulatory properties (Biswas et al., 2024) [22] . Turmeric (Curcuma longa) is widely recognized for its bioactive compound curcumin, which possesses potent antioxidant and anti-inflammatory activities (Kunnumakkara et al., 2017) [27] . Curcumin has been reported to scavenge free radicals, reduce lipid peroxidation, improve immune function, and protect tissues from oxidative damage induced by environmental stressors. Supplementation of turmeric may therefore help maintain physiological stability and improve productivity in heat-stressed buffaloes (Biswas et al., 2024) [22] . Shatavari (Asparagus racemosus) is an important medicinal herb in traditional Indian medicine and is considered a natural adaptogen (Sharma et al., 2021) [31] . It contains steroidal saponins, flavonoids, and polyphenolic compounds that enhance stress tolerance, improve immune competence, and support endocrine balance (Alok et al., 2013) [21] . [32] . The plant exhibits anti-inflammatory, immunomodulatory, and antimicrobial properties (Sahu et al., 2013) [29] . Aloe vera supplementation has been reported to improve digestive efficiency, enhance nutrient absorption, and reduce physiological stress responses (Surjushe et al., 2008) [32] . Its antioxidant activity may help mitigate cellular damage caused by excessive heat exposure (Choi & Chung 2021) [20] . Moringa (Moringa oleifera) is often referred to as a "miracle tree" due to its exceptional nutritional and medicinal value (Sebola, et al., 2019) [30] . The leaves are rich in proteins, essential amino acids, vitamins, minerals, and natural antioxidants such as flavonoids and phenolic compounds (Falowo et al., 2018) [25] . Moringa supplementation has been associated with improved feed intake, enhanced immune function, better antioxidant status, and increased milk production in dairy animals (Khan & Wang 2024). Its ability to counteract oxidative stress makes it a promising dietary intervention for animals subjected to high environmental temperatures (Falowo et al., 2018) [25] .

Although individual studies have reported beneficial effects of these herbal supplements in livestock, information regarding their comparative efficacy in alleviating heat stress in buffaloes remains limited. Therefore, the present study was undertaken to evaluate the effects of dietary supplementation with turmeric, Shatavari, aloe vera, and moringa on productive performance of heat-stressed buffaloes. It was hypothesized that supplementation with these medicinal herbs would improve thermotolerance, reduce stress-induced physiological alterations, and enhance productive performance during periods of elevated environmental temperature.

Materials and Methods

Study area, context and Design

The present study was conducted in Hardoi district, Uttar Pradesh, India. The district is located in the central Gangetic plains and is characterized by a subtropical monsoon climate with hot summers and a humid rainy season. During the summer months (May-June), ambient temperatures frequently range between 40°C and 46°C, accompanied by intense solar radiation and occasional heat waves, resulting in substantial thermal stress in livestock. Relative humidity remains moderate during early summer and increases progressively with the approach of the monsoon. Consequently, the Temperature-Humidity Index (THI) often exceeds the critical threshold for dairy buffaloes, indicating severe heat stress conditions (GOI 2025) [26] . A total of 50 healthy lactating Murrah graded buffaloes in their second parity were selected for the study. All experimental animals were clinically healthy and maintained under similar managemental conditions. At the commencement of the experiment during non-heat stress conditions, the buffaloes produced an average milk yield of 8.2 kg/day, with mean milk fat and solid-not-fat (SNF) contents of 8.3% and 10.5%, respectively. Throughout the experimental period, all animals were provided with a uniform basal ration formulated to meet their maintenance and production requirements. The concentrate mixture consisted of maize/barley/wheat (35 parts), mustard cake or groundnut cake (25 parts), wheat bran (35 parts), mineral mixture (2 parts), common salt (1 part), and dicalcium phosphate (DCP) or limestone powder (2 parts). In addition to the concentrate mixture, animals had free access to clean drinking water and were managed according to standard dairy husbandry practices. A total of 50 lactating Murrah graded buffaloes were randomly allocated into five experimental groups comprising 10 animals each. Group I served as the control and was maintained under ambient environmental conditions without any herbal supplementation. Animals in Group II, Group III, Group IV, and Group V received dietary supplementation of turmeric (Curcuma longa), aloe vera (Aloe barbadensis), shatavari (Asparagus racemosus), and moringa (Moringa oleifera), respectively under ambient environmental conditions. The experiment was conducted for a period of 31 consecutive days during the peak summer season (may 2026), and productive responses associated with heat stress were monitored throughout the study. Ambient temperature (°C) and relative humidity (%) were recorded three times daily,in the morning, afternoon, and evening, using a calibrated digital thermo-hygrometer. The Temperature-Humidity Index (THI) was calculated for each observation using the following equation:

where T represents ambient temperature (°C) and RH denotes relative humidity (%). The daily THI value was obtained by averaging the morning, afternoon, and evening THI measurements. Subsequently, the mean THI for the experimental period was calculated from the daily THI values and used as an indicator of the degree of thermal stress experienced by the animals.

Sample collection

Milk yield was recorded individually for each buffalo on a daily basis throughout the 31-day experimental period. Animals were milked twice daily (morning and evening), and the average daily milk yield was calculated as the mean of the morning and evening milk yields. Group-wise average milk yield was subsequently determined by averaging the daily milk production data of the ten buffaloes within each treatment group over the entire study period. Milk composition analysis was performed at weekly intervals. Representative pooled milk samples, comprising proportional aliquots from the morning and evening milkings, were collected from each animal and analysed for milk fat and solid-not-fat (SNF) percentages using a calibrated electronic milk analyser. Weekly values obtained for each animal were averaged to derive treatment-wise mean fat and SNF percentages for the experimental period. These parameters were used to evaluate the effects of herbal supplementation on milk production and composition in lactating Murrah graded buffaloes subjected to severe heat stress conditions. Fresh leaves, were harvested and spines and outer rind, was removed gel was collected, dried at 50-60°C in hot air oven and then grinded into powder. Mixed with basal ration.

Herbal additive preparation and administration

gm powder/day/animal

All the herbs were fed for 31 consecutive days.

Statistical analysis

Two-way ANOVA was applied to evaluate the effects of herbal supplementation on milk yield, fat percentage, and SNF percentage in heat-stressed buffaloes. The analysis tested the main effects of treatment (herbal supplements) and heat stress category, as well as their interaction, on each response variable. Results were interpreted to determine whether supplementation significantly influenced production and milk composition traits under heat stress conditions. Significance was declared at P< 0.05, and highly significant effects were considered at P< 0.01 and P< 0.0001, as appropriate.

Results and Discussion

The average Temperature-Humidity Index (THI) recorded during the month of May in Hardoi district, Uttar Pradesh, was categorized into four levels, namely low, moderate, high, and severe (Dash et al., 2021) [24] . The effects of treatment, THI level, and their interaction on milk yield, milk fat percentage, and solids-not-fat (SNF) percentage were analysed using a two-way analysis of variance (ANOVA). The statistical model assessed the main effects of dietary treatment and THI category, as well as the interaction between these factors. The results obtained from the analysis are presented below. Milk fat percentage was significantly influenced by both herbal supplementation and THI category. Two-way ANOVA showed a highly significant effect of treatment (P< 0.0001) and THI category (P< 0.0001), whereas the treatment × THI interaction was non-significant (P> 0.05). Among the treatments, Moringa supplementation resulted in the highest fat percentage (8.11±0.05%), followed by Shatavari (7.93±0.04%), Turmeric (7.51±0.07%) and Aloe vera (7.51±0.07%), while the Control group recorded the lowest value (6.86±0.10%). Fat percentage decreased progressively with increasing THI, with the highest values observed under low THI conditions (7.94±0.13%) and the lowest under severe THI conditions (7.35±0.11%). The present findings are consistent with previous reports indicating that phytogenic supplementation enhances milk fat percentage in dairy cattle and buffaloes. Supplementation with Moringa oleifera has been associated with improved rumen fermentation efficiency and increased acetate availability, thereby supporting higher milk fat synthesis in crossbred cows and buffaloes (Dong et al., 2019) [3] . Similarly, improvements in milk composition following Asparagus racemosus (Shatavari) supplementation have been reported in dairy animals, where enhanced lactogenic activity and metabolic modulation were observed (Kumar et al., 2011; Muwal et al., 2020) [5,1] . The significant decline in milk fat percentage with increasing THI is strongly supported by heat stress physiology studies demonstrating that elevated THI reduces dry matter intake, alters rumen volatile fatty acid profiles, and decreases acetate production, ultimately suppressing de novo fatty acid synthesis in the mammary gland (Liu et al., 2017) [2] . Recent large-scale analyses further confirm that heat stress can reduce milk yield and alter milk composition in dairy systems globally, particularly in tropical regions (Rashid et al., 2024) [16] . The non-significant treatment × THI interaction observed in the present study suggests that herbal supplementation improved baseline metabolic efficiency but did not differentially modify milk fat responses under varying thermal stress conditions. It may be due to selection of short period of 31 days. [10] and by Dikmen & Hansen (2009) [8] , who observed significant declines in milk yield with rising THI in dairy cattle. (Bouraoui et al., 2002 & Chen et al., 2024) [7,14] also demonstrated a negative linear relationship between THI and milk production.

The ameliorative effect of herbal supplementation is partially supported by studies reporting improved lactation performance with phytogenic additives. Moringa-based supplementation has been associated with enhanced nutrient utilization and milk yield improvement in ruminants (Fahey, 2005; Taiwo & Aye 2013) [9,6] . However, evidence for herbs such as Shatavari and Aloe vera remains limited and species-dependent, indicating variability across studies. Overall, the present results are consistent with literature confirming heat stress impairment and potential benefits of phytogenic interventions. [13] . Likewise, Shatavari (Asparagus racemosus) has been associated with improved metabolic efficiency and lactational performance due to its phytoestrogenic and adaptogenic properties (Singh et al., 2023) [15] . The positive response observed with Aloe vera supplementation aligns with findings suggesting its bioactive compounds improve gut health and nutrient absorption, indirectly supporting milk solids synthesis (Barman et al., 2017). Similarly, the moderate improvement observed with Turmeric (Curcuma longa) may be attributed to curcumin-mediated antioxidant and anti-inflammatory effects, which mitigate oxidative stress induced by elevated THI (Akbarian et al., 2016) [11] . The decline in SNF percentage with increasing THI is in agreement with the established literature indicating that heat stress reduces feed intake, alters endocrine regulation, and impairs mammary nutrient uptake, ultimately lowering milk solids (Chen et al., 2024) [14] . However, the ability of herbal supplementation to maintain higher SNF levels across THI categories suggests a buffering effect against heat-induced metabolic disturbances. Overall, these results reinforce the hypothesis that phytogenic feed additives enhance thermotolerance and sustain milk quality in heat-stressed buffaloes.

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