Serum biochemical profile of sahiwal cows and osmanabadi goats supplemented with moringa oleifera: A systematic review

Introduction

The serum biochemical profile of a livestock animal is a precise, quantitative reflection of its nutritional status, metabolic homeostasis, and the functional integrity of key visceral organs principally the liver and kidneys. In the context of dairy ruminant production, where the demands of lactation and the peri-parturient period impose severe metabolic stress, periodic biochemical monitoring is not merely a diagnostic luxury but a productive necessity. Disruptions in hepatic protein synthesis, renal nitrogen handling, glucose metabolism, or lipid turnover directly compromise milk yield, reproductive efficiency, and immune competence, particularly during the physiologically critical transition period (Drackley, 1999; Grummer, 1995) [9,15] . Moringa Oleifera Lam. (family Moringaceae), commonly known as the drumstick or miracle tree, is a rapidly growing, drought-tolerant, multipurpose tree indigenous to the Indian subcontinent. Its leaves contain 22.07% crude protein on a dry matter basis and its stem 12-14.5% crude protein values that compare favourably with conventional concentrate ingredients (Shih et al., 2011; El-Esawy et al., 2018) [37,11] . Beyond macronutrients, M. oleifera elaborates a rich array of phytochemicals: flavonoids (quercetin, kaempferol, apigenin), phenolic acids, saponins, condensed tannins, alkaloids, and isothiocyanate glucosinolates, alongside vitamins A, C, E, and key minerals (Fe, Ca, Zn). These constituents collectively confer antioxidant, antiinflammatory, hepatoprotective, hypoglycaemic, lipidlowering, and immunomodulatory properties (Fahey, 2005 [12,4,31,25,2,33,45] . Three masters investigations at the College of Veterinary Science and Animal Husbandry, Anjora, Durg, India, systematically evaluated serum biochemical parameters in: (i) Sahiwal cows supplemented with dried M. oleifera leaves over 35 days (Study 1, Sonkar et al.); (ii) transition-phase Sahiwal cows supplemented with 20% dried M. oleifera stem (DM basis) over 42 days spanning the peri-parturient period (Study 2, Sori et al.); and (iii) Osmanabadi goats supplemented with M. oleifera leaves across four treatment levels over 42 days (Study 3, Kashyap et al.). This review synthesises, integrates, and critically contextualises the serum biochemical data from these three investigations against the existing global literature, with the objective of establishing a definitive evidence base for the biochemical safety and efficacy of M. oleifera supplementation in tropical dairy ruminants.

Experimental Designs and Treatment Protocols

Study 1 (Sonkar et al.) employed a randomised block design with three groups of Sahiwal cows T 0 (control), T 1 (Replacement of 10% DM of concentrate by dried Moringa Oleifera leaves), and T 2 (Replacement of 20% DM of concentrate by dried Moringa Oleifera leaves) over a 35day trial. Blood sampling was conducted at day 0 and day 35. Parameters evaluated: total serum protein, albumin, globulin, BUN, glucose, AST, GGT, LDH, and total triglycerides. Data was analyzed by one-way ANOVA, followed by DMRT for mean comparison, while withintreatment differences were assessed using the paired t-test. Study 2 (Sori et al.) recruited 12 Sahiwal cows (1st-5th parity; BW 300-350 kg) randomly allocated to T 0 (control) and T 1 (20% DM of concentrate replaced by dried M. oleifera stem) from 21 days pre-calving to 21 days postcalving. Blood was collected at three phases: before parturition, at parturition, and after parturition. Parameters assessed: serum glucose, AST, ALT, total protein, albumin, globulin, and total triglycerides. Mean and standard error were calculated following Snedecor and Cochran (1994). Differences between groups were assessed using the independent samples t-test in MS Excel. Study 3 (Kashyap et al.) used Osmanabadi goats distributed across T 0 (control), T 1 , T 2 , and T 3 (Animals were fed green tree leaves along with a 16% CP concentrate. In T 1 , T 2 , and T 3 , soybean meal was replaced by dried Moringa Oleifera leaves at 10%, 20%, and 30%, respectively, while maintaining iso-nitrogenous diets) over 42 days. Parameters assessed: total protein, albumin, globulin, glucose, and BUN, at day 0 and day 42. The data obtained was analyzed using one-way ANOVA following Snedecor and Cochran (1994), and significant differences among means were compared using Duncan's Multiple Range Test (DMRT) as described by Steel and Torrie (1980) [44] .

Serum Protein Fractions 3.1 Total Serum Protein

Total serum protein (TP) is the aggregate of all proteins circulating in the plasma predominantly albumin, globulin, fibrinogen, and acute-phase proteins synthesised by the liver or elaborated by plasma cells. It is the primary index of nutritional protein status, hepatic synthetic capacity, and overall metabolic health. The physiological normal range is 6.8-8.6 g/dl in cattle (Smith, 2002) [38] and 6.0-7.5 g/dl in goats (Kaneko, 1989) [18] .

In Study 1, at day 0, TP was comparable among groups (T 0: 7.06±0. [21] and Damor et al.

(2017b) [7] Similar improvements in nutrient utilization and protein metabolism have recently been reported in dairy cows receiving Moringa supplementation (Sahu et al., 2023; Kholif et al., 2022) [39,23] .. In Study 2 (transition cows), TP showed a phase-dependent pattern. Before parturition, TP was comparable (control: www.extensionjournal.com 5.89±0.34 _ scriptsizei, treatment: 5.79±0.27 g/dl; NS). At parturition, control TP was significantly higher (7.35±0.36 vs. 4.92±0.34 g/dl; P<0.001). This paradoxical finding is physiologically explained by massive immunoglobulin transcytosis from serum to the mammary gland during colostrum formation: moringasupplemented cows, secreting higher-quality, immunoglobulin-richer colostrum, experienced a more pronounced but entirely normal serum TP decline at parturition. By 21 days post-calving, TP had fully recovered to comparable values (T 0: 5.46±0.17, T 1: 5.74±0.18 g/dl; NS), confirming normal protein restitution in the moringa group. In Study 3 (goats), day-42 TP was non-significantly different across groups (T 0 colon [6] .31±0.34, T 1 colon [6] .01±0.18, T 2: 6.84±0.49, T 3: 6.56±0.40 g/dl; P = 0.41-0.91), all within the normal range. The absence of a significant response in goats at maintenance or moderate production likely reflects a lower metabolic protein demand relative to high-producing transition dairy cows, consistent with Elaidy et al. (2017) and Yusuf et al. (2018) [48] .

Serum Albumin

Albumin, synthesised exclusively in the liver and constituting ~60% of total plasma protein, maintains plasma oncotic pressure and serves as the principal carrier for fatty acids, bilirubin, hormones, and drugs. Its concentration is a sensitive, chronic indicator of hepatic synthetic capacity and nutritional protein adequacy. Normal range: 3.0-4.3 g/dl in cattle (Smith, 2002) [38] ; 2.7-3.9 g/dl in goats.

In Study 1, day-35 albumin was significantly higher in T 2 (4.40±0.10g / dl; P<0.05) compared to T 0 (4.06±0.04g / dl), while T 1 (4.25±0.88: g / dl) was intermediate. Within-group analysis confirmed significant increases in T 1 and T 2 scriptstyle (P<0.05) but not T 0 between days 0 and 35. The elevated albumin is attributed to the moringa-derived amino acid supply augmenting hepatic albumin synthetic flux. Although day-35 T 2 albumin slightly exceeds the commonly cited upper limit, elevated albumin is clinically safe and physiologically advantageous, reflecting enhanced hepatic synthetic capacity and improved protein nutrition, not disease (Meel et al., 2018b) [28] . Study 2 yielded perhaps the most clinically significant albumin finding of this review. At parturition, the moringasupplemented group maintained significantly higher albumin (3.25±0.19: g / dl) versus the control (2.38±0.30g / dl; P=0.034). Periparturient hypoalbuminaemia is a wellrecognised predisposing factor for displaced abomasum, peripheral oedema, and hepatic lipidosis in transition dairy cows conditions with major economic consequences (Drackley, 1999) [9] . The significantly higher periparturient albumin in moringasupplemented cows suggests that the sustained high-quality protein supply from M. oleifera stem buffered hepatic synthetic capacity against the albumin demand imposed by colostrum secretion. Pre-calving (3.48± 0.21 vs. 3.25±0.19 g/dl; NS) and post-calving (3.60±0.09 vs. 3.48±0.09 g/dl; NS) values were comparable. El-Esawy et al. (2018) [11] and Meel et al. (2018b) [28] similarly documented higher serum albumin in moringa-fed animals.Recent evidence also indicates improved hepatic protein synthesis and metabolic adaptation in Moringasupplemented dairy cows (Sahu et al., 2023) [39] . In Study 3 (goats), albumin did not differ significantly among groups at either time point (day 42: T 0 3.61±0.26, T 1 3.20±0.30, T 2 3.83±0.23, T 3 3.61±0.29 g/dl; all NS, all within normal range). This speciesspecific absence of response is consistent with Azzaz et al. (2016) [5] , suggesting that the albumin response to moringa-derived protein is modulated by the magnitude of the physiological protein demand greatest in high-producing peri-parturient dairy cows, least in goats at maintenance.

Serum Globulin

Globulin encompass immunoglobulin (IgG, IgM, IgA produced by plasma cells) and hepatic acute-phase proteins (alpha and beta fractions). Their aggregate concentration reflects both the humoral immune status and acute-phase inflammatory response. Normal range in cattle: 3.0-4.9 g/dl (Smith, 2002) [38] . In Study 1, between-group globulin differences were nonsignificant at both time points. However, withingroup analysis revealed a significant increase in T 2 from day 0 (3.25±0.08g / dl) today 35 (3.35±0.03: g / dl;; P<0.05), while T 0 and T 1 showed non-significant trends. All values were within the normal range. The numerical globulin increases in moringa-supplemented groups suggest immunomodulatory benefit plausibly mediated by saponins (which enhance B-lymphocyte-mediated immunoglobulin synthesis) and alkaloids (which possess adjuvant-like properties) present in M. oleifera leaves (Wina et al., 2005) [46] . Ahmad et al. (2017) corroboratively documented nonsignificantly higher globulin in moringa-supplemented buffalo calves. Recent studies suggest that Moringa bioactive compounds may enhance immune responsiveness and antioxidant status in ruminants (Abd El-Hack et al., 2023; Frontiers Nutrition Review Group, 2025) [1,14] In Study 2, globulin showed the expected periparturient variation. The dramatic numerical difference at parturition (control: 4.97±0.54 vs. treatment: 1.70±0.24 g/dl; NS due to high within-group variance) reflects the colostrum-transfer physiology discussed under total protein. Post-calving globulin recovery was comparable and within the normal range in both groups. In Study 3, globulin was nonsignificantly different across goat treatment groups and time www.extensionjournal.com points (day 42: T 0 2.70±0.16, T 1 2.90±0.11, T 2 3.01±0.24, T 3 2.94±0.19 g/dl; NS), consistent with Mahmoud (2013) [27] and Meel et al. (2018b) [28] .

Renal Function: Blood Urea Nitrogen

Blood urea nitrogen (BUN) is the primary nitrogenous endproduct of the hepatic urea cycle, generated from the catabolism of dietary and endogenous amino acids. In ruminants, it also reflects rumen urea recycling a physiologically beneficial process in which blood urea reenters the rumen via salivary secretion and direct epithelial transfer to serve as non-protein nitrogen for rumen microorganisms. Elevated BUN beyond the reference range (20-30 mg/dl in cattle; 10-30 mg/dl in goats; Kaneko et al., 2008) [19] in isolation may indicate excess dietary protein, while concomitant elevation of both BUN and creatinine signals primary renal failure. In Study 1, BUN at day 35 was non-significantly different across groups (T 0: 24.45±0.62, T 1: 25.47±0.25, T 2: 24.87±1.20 mg/dl; all within normal range; NS). The slight numerical increase in moringa groups is the expected physiological consequence of increased dietary protein catabolism not renal impairment. Importantly, this modest urea elevation stimulates rumen urea recycling, improving microbial protein synthesis efficiency (Yusuf et al., 2018) [48] . In Study 3, BUN on day 42 was comparable across goat treatment groups (T 0: 47.18±2.9, T 1: 46.85±3. [48] in goats, and with Haridas (2018) [16] in growing sheep. The consistent maintenance of BUN within physiological reference ranges across both species and all doses definitively confirms renal safety of M. oleifera supplementation. Comparable findings have been reported in recent goat and cattle studies supplemented with Moringa foliage (Afzal et al., 2022; Din et al., 2026) [3,8]

Carbohydrate Metabolism: Serum Glucose

In ruminants, blood glucose is derived predominantly from hepatic gluconeogenesis primarily from propionate, lactate, glycerol, and glucogenic amino acids rather than from intestinal glucose absorption. Normal range: 45-75 mg/dl in cattle; 50-75 mg/dl in goats (Kaneko et al., 2008) [19] . Hypoglycaemia during early lactation is a primary trigger of ketosis and negative energy balance. In Study 1, serum glucose at day 35 was within normal range and non-significantly different across groups (T 0: 52.15±0.73, T 1: 53.32±1.42, T 2: 52.83±3.27 mg/dl; NS). The slight non-significant increase in moringasupplemented groups is attributed to chlorogenic acid facilitating glucose bypass absorption from the small intestine (Khalel et al., 2014) [21] . Results agree with Zeng et al. (2018) [49] in Holstein dairy cows. Study 2 presented a more nuanced glucose response. Before parturition, serum glucose was significantly lower in the moringa-supplemented group (58.00±2.38 mg/dl) versus control (73.50±2.55 mg/dl; P = 0.001). This is mechanistically explained by: (a) chlorogenic acid inhibiting intestinal glucose-6-phosphatase, reducing postabsorptive glucose release; (b) flavonoids inhibiting αamylase and α-glucosidase; and (c) the higher acetate-topropionate ratio of moringa stem fermentation products reducing hepatic gluconeogenic substrate supply (Farooq et al., 2007) [13] . Crucially, even the lower pre-partum glucose value (58 mg/dl) remained within the normal range (45-75 mg/dl), confirming this was a modulated physiological response and not pathological hypoglycaemia. Post-calving values were non-significantly different between groups (P = 0.205). In Study 3, all glucose values were within the normal range and non-significantly different across goat groups at day 42 (T 0: 60.8±1.4, T 1: 61.44±3.9, T 2: 58.54±3.8, T 3: 59.38±4.30 mg/dl; NS). The slight decrease in T 2 from its initial value reflects flavonoid-mediated glycaemic modulation, consistent with Babekar and Abdalbagi (2015) [6] . The collective glucose data across all studies confirm that M. oleifera supplementation does not induce pathological hypoglycaemia or hyperglycaemia at any evaluated dose in either ruminant species.Recent investigations attribute this glycaemic modulation to chlorogenic acid and polyphenolic compounds present in Moringa leaves (Louisa et al., 2022; Din et al., 2026) [26,8] . [19] . All values within normal physiological range. NS = Nonsignificant; \\ P<0.01.

Hepatic Enzyme Markers 6.1 Aspartate Aminotransferase (AST)

Aspartate aminotransferase (AST; EC 2.6.1.1) is a pyridoxal phosphate-dependent transaminase distributed in hepatocytes, cardiac myocytes, skeletal muscle, and erythrocytes. Its release into circulation indicates loss of cellular membrane integrity specifically hepatocellular damage when other hepatic markers co-elevate (Molander et al., 1957) [29] . Normal range in cattle: 78-132 U/L (Kaneko et al., 2008) [19] . In Study 1, AST values were within the normal range throughout. At day 0: T 0 79.25±6.66, T 1 86.28±3.59, T 2 88.21±6.68 U/L (NS). At day 35: T 0 85.03±7.16, T 1 83.65±5.43, T 2 84.66±7.70 U/L (NS). Notably, T 0 AST increased slightly from day 0 to day 35, while T 1 and T 2 values were numerically stable or declining a pattern consistent with the hepatoprotective action of quercetin and other flavonoids in M. oleifera, which stabilise hepatocyte membranes and inhibit lipid peroxidation (Nascimento et al., 2014; Anwar et al., 2007) [31,4] . These results concur with Khalel et al. (2014) [21] and El-Esawy et al. (2018) [11] . Recent studies further support the hepatoprotective role of www.extensionjournal.com Moringa through antioxidant-mediated stabilization of hepatocyte membranes (Pareek et al., 2023; Abd El-Hack et al., 2023) [33,1] . In Study 2 (transition cows), AST was non-significantly different between groups at all three phases: before parturition (72.86±3.64vs. 66.70±4.70U / L; P=0.325), at parturition (82.69±9.67vs. 78.31±6.94U / L; P =0.72), and after parturition (89.99±3.81 vs. 97.67±8.22 U/L; P = 0.416). All values were within the normal range. The physiological post-calving AST increase in both groups reflects parturient uterine involution and early-lactation hepatic lipid mobilisation not primary hepatocellular disease. Normal AST in all moringa-supplemented animals unambiguously confirms hepatocellular integrity and the absence of hepatotoxicity.

Gamma-Glutamyl Transferase (GGT)

GGT (EC 2.3.2.2) is concentrated in the liver's biliary canalicular membrane and is considered the most sensitive serum marker of biliary obstruction, cholestasis, and early hepatocellular injury in dairy cattle. Chronically elevated serum GGT may also indicate hepatic fatty infiltration or toxic hepatopathy (Moore, 1997) [30] . Normal range: 6.1-17.4 U/L (Kaneko et al., 2008) [19] . In Study 1, GGT concentrations were within normal range at all time points, with non-significant betweengroup differences. Day 0: T 0 11.67±1.36,, T 1 11.82±1.88, T 2 11.39±3.38 U/L (NS). Day 35: T 0 11.06±1.17, T 1 10.72±1.22, T 2 11.09±1.45U / L (NS). All three groups showed a non-significant decreasing trend from day 0 to day 35, with moringa-supplemented groups exhibiting numerically lower values. This hepatoprotective pattern is attributable to the membrane-protective action of M. oleifera flavonoids particularly their free radical scavenging activity that targets oxidants threatening biliary canalicular membrane permeability (Sharifudin et al., 2012) [36] . Normal GGT in all supplemented animals conclusively rules out biliary toxicity or cholestatic hepatopathy from M. oleifera supplementation. Similar observations have been reported in modern livestock nutrition studies utilizing Moringa-based feed additives (Abdoun et al., 2023; Su et al., 2023) [2,45] . [19] . In Study 1, LDH concentrations were within the normal range throughout, with non-significant betweengroup differences at both time points. Day 0colonT 0 805.10±24.04., T 1 813.98±25.36, T 2 811.67±26.46 U/L (NS). Day 35: T 0 809.15±28.05, T 1 802.92±24.46, T 2 808.12±25.17 U/L (NS). The T 0 control showed a slight non-significant increase in LDH from day 0 to day 35, while both moringasupplemented groups showed non-significant decreases mirroring the AST and GGT patterns and pointing to a generalised antioxidant-mediated tissue-protective effect of M. oleifera phytochemicals (Johnson-Davis and McMillin, 2010) [17] . Normal LDH values across all groups confirm efficient multi-organ function and the complete absence of cytotoxic effects from M. oleifera supplementation. Recent reviews likewise indicate that Moringa supplementation does not adversely affect biochemical indicators of tissue integrity in ruminants (Din et al., 2026; Abd El-Hack et al., 2023) [8,1] [19] . All values within normal limits throughout. 7. Lipid Metabolism: Serum Triglycerides Total serum triglycerides (TG) in ruminants originate primarily from hepatic de novo synthesis (using acetate, propionate, and glycerol) and from lipolysis of adipose tissue during negative energy balance periods. Elevated serum TG is associated with impaired hepatic lipid export and fatty liver risk a major periparturient metabolic disorder in dairy cows. Normal range in cattle: 0-14 mg/dl (Kaneko et al., 2008) [19] . The triglyceride data from Study 1 (Sonkar et al.) represent one of the most statistically and metabolically significant findings of this review. At day 0, TG was comparable across groups (T 0: 11.75±0.77, T 1: 11.87±0.83, T 2: 11.13±1.32 mg/dl; NS). By day 35, T 2 exhibited a significantly lower TG (8.27±0.79mg / dl; P<0.05) compared to T 0 (11.08±0.88mg / dl), with T 1 intermediate (9.13±0.40 mg/dl). Within-group analysis revealed highly significant (P<0.01) TG reductions in both T 1 and T 2 from day 0 to day 35, while T 0 showed no significant change. All values remained within the normal range throughout. This dose-dependent triglyceride-lowering effect is mechanistically grounded in the phenolic chemistry of M. oleifera. Saponins, condensed tannins, and flavonoids form insoluble complexes with intestinal cholesterol and triglycerides via hydrophobic interactions and hydrogen bonding, preventing their emulsification, micelle formation, and subsequent intestinal absorption. The resulting insoluble lipid-phytochemical complexes are excreted in faeces, reducing portal TG delivery to the liver (Saxena et al., 2013; Adriani et al., 2014) [35] . Additionally, quercetin has been reported to inhibit pancreatic lipase activity, further reducing triglyceride hydrolysis and absorption, while upregulating hepatic lipoprotein lipase to enhance TG clearance from circulation. These findings are consistent with Kholif et al. (2016) [22] , who documented lower serum TG in moringa-leaf-meal-supplemented Anglo-Nubian goats. Recent evidence confirms the hypolipidaemic effects of Moringa bioactive compounds through modulation of lipid metabolism and antioxidant pathways (Sahu et al., 2023; Louisa et al., 2022) [39,26] . In Study 2 (transition cows), TG before parturition was significantly higher in the moringa-supplemented group (85.30±3.61g / dl) versus control (59.10±3.88g / dl; www.extensionjournal.com P=0.0005). This reflects heightened adipose mobilisation fuelling the elevated energy demands of both late pregnancy and the augmented pre-partum milk protein synthesis in the moringa group. Post-calving TG was numerically lower in the moringa group (70.23±12.80 g/dl) versus control (83.65±5.59g / dl; P=0.359; NS), consistent with the gradual expression of M. oleifera's lipid-lowering phytochemistry over time. All post-calving values were within the normal range. [19] . \ P<0.05 (between-groups at day 35); \\ scriptstyleP<0.01 (withingroup day 0 vs. day 35). NS = Non-significant. Table 6 presents the complete master biochemical profile from Study 1 (Sonkar et al.), encompassing all nine parameters evaluated at both time points. This integrated view permits comprehensive assessment of the overall metabolic health trajectory and the systematic influence of moringa leaf supplementation across multiple biochemical axes. [38] and Kaneko et al. (2008) [19] . All values within physiological reference limits throughout the experimental period.

Lactate

Integrated Biochemical Profile Master Summary Table

Mechanistic Basis of Biochemical Effects

The constellation of biochemical effects documented across the three investigations elevated protein fractions, reduced triglycerides, glycaemic modulation, and maintenance of all hepato-renal safety markers can be coherently unified through four principal mechanistic pathways:  Enhanced protein supply and hepatic synthetic capacity: The 22.07% CP (leaves) and 12-14.5% CP (stem) content of M. oleifera provides rumendegradable nitrogen for microbial protein synthesis and rumen-undegradable protein enriching duodenal amino acid flow. This dual protein supply elevates hepatic amino acid availability, directly stimulating albumin and total protein synthesis in a dose-responsive manner (Soliva et al., 2005). The magnitude of the albumin response scales with physiological protein demand greatest in peri-parturient dairy cows, negligible in goats at maintenance (Kholif et al., 2022; Sahu et al., 2023) [23,39] [13,21,8,25] .

Conclusion

This systematic review of serum biochemical data from three independent, rigorously conducted masters investigations provides a comprehensive and definitive evidence base for the biochemical safety and efficacy of Moringa Oleifera supplementation in Sahiwal dairy cows www.extensionjournal.com and Osmanabadi goats. The following conclusions are unambiguously supported by the collective data: Serum total protein and albumin were significantly elevated (P<0.05) at higher M. oleifera doses in Sahiwal dairy cows. Albumin was significantly protected against periparturient decline in transition cows (P = 0.034 at parturition). All values remained within physiological norms, confirming nutritional benefit without hyperviscosity risk.  Serum globulin showed numerically higher concentrations in supplemented groups, with a significant within-group increase in the high-dose cow group (P<0.05), suggesting immunomodulatory benefit. Between-group differences were non-significant, ruling out pathological hypergammaglobulinaemia. [39,3,8] .

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