Chemical composition of feeds and herb
The chemical composition of wheat straw, concentrate mixture and green maize fodder (Table 1) was in accordance with
Chaudhary et al., (2021). Chemical composition of
T. cordifolia,
W. somnifera and
A. racemosus was within the normal range as reported by earlier workers
(Chavan et al., 2021; Verma et al., 2024).
Phytochemicals in herbs
The substantially higher TPC and TFC values were found in
T. cordifolia relative to
W. somnifera and
A. racemosus (Table 2). These values were within the reported standard ranges
(Royani et al., 2023). In contrary,
Shwetha et al., (2016) reported lower levels of TPC and TFC in
T. cordifolia, while
Udayakumar et al., (2010) observed higher TPC and TFC in
W. somnifera. Such discrepancies in phyto-constituent levels may be ascribed to variations in extraction protocols as well as ecological factors, including soil type, climate, sunlight exposure and geographical variation (
Nobossé et al., 2018). The IC
50 values were higher (P≤0.001) in
A. racemosus, followed by
W. somnifera and
T. cordifolia, indicating that
T. cordifolia exhibited the strongest antioxidant potential among the three, as lower IC
50 values signify enhanced antioxidant activity
(Olugbami et al., 2014).
Feed intake
DM and OM intake were higher (P<0.01) in T1 and T2 groups, consistent with earlier reports that increased energy density of rations or concentrate levels enhanced DM intake in ruminants
(Hailu et al., 2011; Tufarelli et al., 2011; Chaudhary et al., 2020). Moreover, supplementation of
A. racemosus @ 125 and 150 mgkg
-1 BW led to higher DM intake in Sahiwal heifers
(Kumawat et al., 2017). Mir et al., (2015) reported significant increase in DMI in lactating Murrah buffaloes fed with
T. cordifolia @ 120 gd
-1. The elevated DMI was likely associated with increased concentrate intake, leading to greater availability of fermentable carbohydrates and protein that promoted more efficient rumen fermentation.
Metabolic profile
Early post-partum cows frequently suffer with negative energy balance due to enhanced energy requirements and reduced DMI
(Bradford et al., 2015). The noteworthy decline of NEFA levels in T2 group (P=0.002) aligned with that of
Sharma et al., (2014), who reported decreased NEFA levels in Karan Fries cows given
W. somnifera,
A. racemosus and
T. cordifolia mixture (200-250 mg kg
-1 BW). These herbs could inhibit lipolysis, enhance lipid metabolism and improve metabolic status of cows (
Hashemzadeh-Cigari et al., 2015). Serum urea and cholesterol levels were uninfluenced and were coherent with
Sharma et al., (2018). However,
Kumar et al., (2014) observed reduced cholesterol in Karan Fries cows with
A. racemosus (100 mgkg
-1 BW). Overall, the herbal feed additive with high plane of nutrition maintained stable homeostasis without altering the blood biochemical profile.
Serum hormonal profile
There was a substantial (P<0.05) rise in serum T3, T4 and progesterone (Table 3). Previous reports
(Patel et al., 2016; Rajneesh et al., 2020) showed that supplementation of
T. cordifolia based herbal mixture positively affected serum T4 level in buffalo calves. The positive effect of addition of herbal feed additive could be explained by thyroid regulating and stimulating effect of
T. cordifolia and Withaferin A in
W. somnifera (
Tewari and Tewari 2020) and the higher conception rate in T2 group. The reduced serum cortisol level was likely due to reduction in adrenocorticotropic hormone caused by
W. somnifera and
A. racemosus (Singh et al., 2023).
Antioxidant profile
The present results exhibited higher (P≤0.001) SOD, CAT, GPx and TAOC, along with a reduced (P=0.04) LPO in T2 group (Fig 1). These findings are compatible with
Vaswani et al., (2022) and
Oviya (2023) who reported elevated SOD activity while reduced LPO levels in Sahiwal heifers and increased SOD, GST and GPx activity in goat kids by supplementation of
T. cordifolia at 1% of DMI and herbal mixture (
W. somnifera and
A. racemosus; 2:1) @150 mgkg
-1 BW, respectively.
The antioxidant consequences of
W. somnifera and
A. racemosus are attributed to its bioactive constituents, notably withanolides (Sitoindosides VII-X), withaferin A, racemofuran, asparagamine A and racemosol (
Ojha and Arya, 2009;
Wiboonpun et al., 2004). Moreover, the flavonoids present in these herbs regulate the expression of gamma-glutamylcysteine synthetase
(Moskaug et al., 2005). Therefore, the beneficial effects may result from the synergistic action of phytobiotics in the herbal mixture.
CMI response
The cows in T2 group showed a noticeably enhanced (P≤0.001) CMI response (Fig 2). This suggests a potent immunomodulatory effect of the herbal feed additive which is in line with
Mallick and Prakash (2011) who stated considerably greater lymphocyte counts in
T. cordifolia fed (@ 60 gd
-1 for 45 d before parturition and @ 120 g d
-1 for 45 d post-calving) Karan Fries cows. The immunomodulatory action of herbal feed additive is primarily endorsed to its active components like α-D-glucan, Withaferin A, 3-β-hydroxy-2, 3-dihydro withanolide F, Shatavaroside A and Shatavaroside B
(Thakur et al., 2021). Moreover, carotenoids existing in both
A. racemosus and
W. somnifera have been stated to enhance the proliferation of T and B lymphocytes, stimulate macrophage activity and promote the production of pro-inflammatory cytokines
(Milani et al., 2007). Thus, the synergistic action of these phytoconstituents likely contributed to the substantially elevated CMI response.
Lactation performance
Supplementation in the T1 and T2 groups significantly (P≤0.001) increased daily and total milk yield (Table 4), 4% FCM, ECM (kg day
-1) and fat and protein yields (g day
-1), without affecting milk composition. Our results line up with
Choudhary et al., (2024) who reported increased milk production, similar milk composition and improved 4% FCM in milching cows with
T. cordifolia and
A. racemosus feeding.
Dori et al. (2026) and
Prasanna et al. (2023) also reported similar milk composition in cows fed dried moringa stem. In contrary,
Sharma et al., (2017) observed similar milk yield in crossbred cows fed
T. cordifolia stem powder. The role of dietary energy and protein in supporting lactation is well established with enhanced production linked to increased availability of rumen-fermentable carbohydrates, degradable N and efficiency of microbial protein synthesis
(Zhou et al., 2015). Thus, the improved milk yield was likely due to higher concentrate feeding.
Furthermore, the enhanced milk DPPH activity in T-2 group was supported by
Stobiecka et al., (2023), who witnessed enhanced antioxidant potential following herbal administration. Herbal feed additives are positively correlated with DPPH radical scavenging activity
(Aryal et al., 2019).
Reproduction performance
The cows in the T2 group exhibited an earlier onset of estrus (P=0.043), a higher conception rate (P≤0.01) and required significantly (P=0.01) less artificial inseminations per conception (Fig 3). These findings align with
Kumar et al., (2012) who documented earlier onset of estrus and improved conception rates in post-partum cows provided
A. racemosus @ 125-200 mgkg
-1 BW. Similarly,
Muwal et al., (2020) reported reduced service periods, services per conception and earlier estrus induction in dairy cows
. Likewise,
Mallick and Prakash (2011) reported earlier onset of estrus in post-partum Karan Fries cows by dietary inclusion of
T. cordifolia stem powder @ 60 g d
-1 which was mainly due to its immunomodulatory properties that promote early resumption of ovarian cyclicity.
Singh et al., (2022) reported reduced service periods and improved conception rates in Sahiwal cows given polyherbal mixture containing
W. somnifera @ 20 g d
-1 from 3 d pre-partum to 21 d post-partum. Previous studies have confirmed that
A. racemosus supports postpartum uterine involution and reproductive health through stimulation of epithelial cell growth and ovarian function. Thus, the presence of bioactives in herbal feed additive could have enhanced the reproductive performance by supporting follicular development and hormonal balance
(Kumar et al., 2014).
