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volume 45 safe, sustainable, and innovative dairy & food systems for one health : 42-50,   Doi: 10.18805/ajdfr.DR-2468

Assessment of The Digestive Efficacy of a Nutritional Supplement in Bovines

M
M.P. Sakhare1
S
S.P. Risbud2,*
M
M.F.M.F. Siddiqui1
P
P.C. Rathi2
C
C.L. Rathi2
V
V.K. Munde1
A
A.M. Chappalwar1
A
A.M. Syed1
S
S.R. Shaikh1
1Department of Veterinary Epidemiology and Preventive Medicine College of Veterinary and Animal Sciences, Maharashtra Animal and Fishery Science Unviersity, Parbhani-431 402, Maharashtra, India.
2Advenza Global limited (formerly known as Advanced Vital Enzymes Pvt. Ltd.), Wagle Estate, Thane(W)-400 604, Maharashtra, India.
Cite article:- Sakhare M.P., Risbud S.P., Siddiqui M.F.M.F., Rathi P.C., Rathi C.L., Munde V.K., Chappalwar A.M., Syed A.M., Shaikh S.R. (2026). Assessment of The Digestive Efficacy of a Nutritional Supplement in Bovines . Asian Journal of Dairy and Food Research. 45: 42-50. doi: 10.18805/ajdfr.DR-2468.

ABSTRACT

Background: Digestive disorders in cattle impair rumen function, feed utilization and productivity. Nutritional strategies such as probiotics, enzymes and herbal extracts may enhance ruminal health. This study evaluated the efficacy of a rumenotonic bolus containing probiotics, enzymes and herbal extracts in managing ruminal disorders in bovines.

Methods: A clinical trial was conducted on 18 lactating cows with ruminal disorders at the Department of Veterinary Medicine, Parbhani (February-June 2025). Animals were divided into three groups: Group I (healthy control), Group II (anorexia treated with standard therapy) and Group III (anorexia treated with rumenotonic bolus, 2 boli BID for 5 days). Clinical, ruminal, hematological, fecal and milk parameters were assessed on days 0, 5 and 7.

Result: Ruminal motility, pH and fermentation parameters improved significantly in treated groups, with the greatest response in Group III. Hematological indices normalized and fecal consistency improved, with faster resolution of diarrhoea in Group III. Milk yield, fat and solids-not-fat increased significantly, while protein remained stable. Group III showed the most rapid recovery (Days 3-5), compared to slower improvement in Group II (Days 5-7). Overall, the rumenotonic supplement enhanced ruminal function and clinical recovery more effectively than standard treatment.

INTRODUCTION

India possesses one of the largest bovine populations globally, with 192.49 million cattle and 109.85 million buffaloes (Government of India, 2019). Maharashtra ranks fifth in cattle and seventh in buffalo population nationally. Cattle and buffaloes constitute 35.94% and 20.45% of India’s livestock population, respectively, highlighting their vital role in the rural economy and dairy sector. However, productivity is often compromised by health disorders, particularly digestive ailments, among which ruminal disorders are most prevalent.
       
In ruminants, conditions such as anorexia, tympany and indigestion are characterized by reduced feed intake, altered ruminal pH, decreased motility and reduced protozoal counts. Simple indigestion commonly arises from sudden dietary changes, poor-quality fodder, imbalanced roughage-to-concentrate ratios, or inadequate water intake (Biswal et al., 2016). These disturbances may lead to ruminal acidosis or alkalosis, depending on fermentation patterns (Constable et al., 2017). Additionally, indigestible feed accumulation and poor feeding practices further aggravate ruminal dysfunction.
       
The rumen, the largest and most vital digestive organ of ruminants, functions as a fermentation chamber housing a complex microbial ecosystem that maintains physiological homeostasis. The rumen is responsible for fermenting fibrous feeds into volatile fatty acids, the primary energy source for ruminants (Moran, 2005; Smith and Jones, 2009). Disruption of this microbial balance adversely affects nutrient utilization and predisposes animals to metabolic and digestive disorders such as acidosis, diarrhea, ketosis, mastitis and laminitis (Asrat et al., 2015; Pan, 2016; Zhao et al., 2023). Maintaining ruminal health is therefore essential for optimal productivity (Beauchemin, 2002).
       
The rationale for this study is based on the need for effective nutritional interventions that can restore ruminal microbial balance, improve digestion and enhance recovery from digestive disorders. Probiotics and exogenous enzymes have gained importance as nutritional interventions to restore ruminal function. These supplements enhance microbial balance, improve digestion and increase feed efficiency (Anadon et al., 2006; Uyeno et al., 2015). Enzymes, amino acids and herbal additives further support nutrient utilization and recovery during anorexia (Arowolo et al., 2018; Mazinani et al., 2022; Chaudhary et al., 2021; Ramdani et al., 2025).
       
Considering the high incidence and economic impact of ruminal disorders, the present study was undertaken with an objective to evaluate the efficacy of a multi-component nutritional supplement (bolus) containing probiotics, enzymes, amino acids and herbal extracts in improving ruminal function and managing ruminal disorders in bovines.

MATERIALS AND METHODS

Study area
 
The present clinical trial was conducted on 18 lactating cows presented to the Department of Veterinary Medicine, Veterinary Clinical Complex (VCC), Instructional Livestock Farm Complex (ILFC), College of Veterinary and Animal Sciences, Maharashtra Animal and Fishery Sciences University (MAFSU), Parbhani and nearby veterinary dispensaries in Parbhani district during February to June 2025. The study evaluated the effect of a nutritional supplement on ruminal disorders in lactating cows, with emphasis on rumen function, milk yield and milk composition.
 
Test product
 
The investigational product is a Rumenotonic with digestive enzymes, amino acids, probiotic and herbal extract supplement containing Amylase, Cellulase, Xylanase - Lysine-DL, Methionine, Histidine, Leucine, Valine, Threonine, Lactobacillus acidophilus, Bacillus coagulans, Saccharomyces boulardii, Aspergillus oryzae, ginger powder and soyabean meal, supplied by Advenza Global Limited in the form of a bolus.
 
Study design
 
Lactating cows diagnosed with ruminal disorders, including simple indigestion, acidic indigestion, alkaline indigestion and anorexia of unknown origin, irrespective of age and breed, were selected based on detailed history and thorough clinical examination. Observations were recorded on day 0 (before treatment), day 5 and day 7 (after treatment).
       
Eighteen cattle were divided into three groups (n = 6 each): Group I comprised healthy animals maintained under identical feeding and management conditions (control); Group II included clinical cases of anorexia of unknown origin treated with a standard protocol and Group III consisted of similar anorexic cases treated with a nutritional supplement administered as two boli twice daily for five days.
 
Sample size
 
The sample size was determined based on feasibility, ethical considerations and availability of animals with similar clinical conditions. A total of 18 animals were included and divided into three groups (Group I, II and III; n = 6 each). This size was considered sufficient to detect biologically meaningful differences while minimizing animal use, consistent with previous studies (Charan and Kantharia, 2013).
 
Clinical and physiological parameters
 
A detailed anamnesis and clinical examination were conducted for all animals. Heart rate was recorded by auscultation over the left thoracic region (3rd-5th intercostal space) and expressed as beats per minute. Respiratory rate was measured by observing nostril movement and thoracic auscultation, expressed as breaths per minute. Rectal temperature was recorded using a digital thermometer (°F). Rumen motility was assessed by palpation and auscultation of the left paralumbar fossa and expressed as contractions per two minutes. Bloat and anorexia were graded using standardized 0-3 scoring scales.
 
Rumen fluid collection and examination
 
Rumen fluid was collected aseptically from the left paralumbar fossa using an 18G, 6-inch spinal needle and extraction pump (Fig 1 and Fig 2). Physical characteristics (colour, odour, consistency) were visually assessed (Fig 3 and Fig 4). pH was measured using indicator paper (HiMedia Laboratories) and categorized as mild (5.5-6.5), moderate (4.5-5.5) and severe (4.0-4.5) acidity (Fig 5 and Fig 6). Total protozoal count was estimated using the method of Moir (1951) and expressed as ×105 /ml.

Fig 1: Collection of ruminal fluid by stomach tube.



Fig 2: Collection of ruminal fluid from left paralumbar fossa.



Fig 3: Brownish green rumen fluid.



Fig 4: Yellowish green rumen fluid.



Fig 5: Normal rumen fluid pH hematological, fecal and milk analysis.



Fig 6: Alkaline pH of rumen fluid.


       
Blood samples were collected on day 0, 5 and 7 into EDTA (3.5%) vacutainers and analyzed using an automatic hematology analyzer (Spincell 5). Fecal examination assessed colour, odour, consistency and diarrhea. Milk yield and composition (pH, fat, protein, Solid -Non-Fat) were recorded and analyzed using a LactoScan analyzer (Fig 7 and Fig 8).

Fig 7: Automatic milk analyzer.



Fig 8: Estimation of milk samples.


 
Statistical analysis
 
Data obtained were subjected to statistical analysis following the methods described by Snedecor and Cochran (1994). Analysis was performed using Completely Randomized Design (CRD) with SPSS version 16. Results were expressed as mean ± standard error (SE).

RESULTS AND DISCUSSION

The present study evaluated efficacy of a multi-component nutritional supplement on ruminal disorders in lactating cows by assessing clinical, physiological, ruminal, haematological, fecal and milk production parameters. Observations recorded at different time intervals (day 0, 5 and 7) provided insights into the progression and recovery pattern in treated animals compared to controls. The findings are discussed in relation to improvements in rumen function, microbial activity and overall health status.
 
Physiological and clinical examination
 
The physiological and clinical parameters in cows affected with simple indigestion showed anorexia, mild depression, reduced feces which were initially dry and and later became softer, voluminous and malodorous consistent with Kasaralikar et al., (2014); Syed et al., (2017) and Constable (2017). The heart rate, respiration rate and rectal temperature remained within normal limits and comparable to controls as presented in Table 1. Post-treatment, no significant changes (p<0.05) occurred except a slight heart rate increase in Group III (p = 0.048), remaining within normal range, similar to previous reports (Kasaralikar et al., 2014; Syed et al., 2017; Constable et al., 2017).

Table 1: The Physiological parameters of animals before and after treatment.


       
Ruminal motility (Table 1) showed non-significant variation in Group I, while indigestion-affected cows in Groups II and III exhibited reduced baseline motility on Day 0, consistent with earlier reports (Walia et al., 2011; Kasaralikar et al., 2014). Group II showed significant improvement during treatment, whereas Group III demonstrated a highly significant progressive increase from 1.33±0.21 to 2.83±0.17 (p = 0.000), with Day 5 and Day 7 values significantly higher than baseline. This restoration approached normal ruminal motility (~3 contractions/2 minutes) as reported by Chakrabarti (2014) and Constable et al., (2017). Among all groups, Group III showed the greatest improvement.
       
In Group I, the left paralumbar fossa was normal, whereas mild distension was observed in a few cows (n = 5) in Groups II and III and the remaining (n = 13) showed a firm, doughy rumen. These findings agree with Constable et al., (2017). Post-treatment, all cows in Groups II and III showed normal paralumbar fossa, indicating resolution of distension, consistent with Walia et al., (2011) and Hussain and Uppal (2012).
       
Appetite remained normal in Group I, while reduced appetite was observed in all affected cows before treatment, as also reported by Constable et al., (2017). Post-treatment, normal appetite was restored in all cows of Groups II and III, indicating effective clinical recovery.
 
Rumen fluid examination
 
Physical examination
 
Healthy cows (Group I) exhibited greenish to yellowish-brown rumen fluid with aromatic odour and viscous-to-thick consistency (Handekar et al., 2010; Kasaralikar et al., 2014). In simple indigestion, it changed to yellowish-green to light brown with occasional ammoniacal odour and watery to semi-solid consistency (Handekar et al., 2010). Post-treatment in Groups II and III, parameters normalized, indicating restoration of rumen function (Waghmare et al., 2009). Observations are presented in Table 2.

Table 2: The physical rumen fluid examination parameters before and after treatment.


 
Physicochemical and microbiological findings
 
The mean rumen liquor pH remained within the normal range in control cows (Chakrabarti, 2014; Constable et al., 2017), while significantly higher values (p<0.01) were observed in Groups II and III before treatment, indicating ruminal alkalinization (Biswal et al., 2016). Post-treatment, pH declined significantly in Groups II and III, reaching near-normal levels by Day 5 and Day 7.
       
Protozoal counts in Group I showed a mild, non-significant increase, while Groups II and III had significantly lower counts pre-treatment with marked post-therapy improvement, especially in Group III (p<0.05), consistent with Mohan et al., (2015). This recovery is closely linked to rumen pH, as protozoa are highly pH-sensitive (Granja-Salcedo et al., 2016) and ruminal acidosis reduces microbial balance through excess volatile fatty acids (TVFA) and lactic acid formation (Biswal et al., 2016). Protozoa play a key role in fermenting carbohydrates into acetic and butyric acids (Chakrabarti, 2014), so their restoration indicates improved fermentation efficiency (Petrovski, 2016).
       
TVFA levels remained normal in Group I (6-12 mEq/dl; Chakrabarti, 2006), while Groups II and III showed significant changes (p<0.01), with Group III exhibiting increased levels (Bhatt et al., 2009). Ammonia nitrogen remained stable in Group I but improved significantly in Group III, indicating enhanced microbial protein synthesis (Shen et al., 2023). Mean values are presented in Table 3.

Table 3: The rumen fluid examination parameters before and after treatment.


 
Hematological findings
 
The results of hematological examination of all groups are presented in Table 4. Haemoglobin and total erythrocyte count increased significantly in Groups II and III, while total leukocyte count decreased significantly, indicating recovery. Other leukocyte parameters remained mostly unchanged. These findings align with earlier studies (Kasaralikar et al., 2014; Syed et al., 2017).

Table 4: Hematological parameters of all groups before and after treatment.


 
Fecal examination
 
Fecal examination showed normal greenish-brown, semisolid feces in Group I (Biswal et al., 2016). In Groups II and III, feces were yellowish-brown, loose and foul-smelling, indicating impaired fermentation (Biswal et al., 2016; Petrovski, 2016). Post-treatment, Group II partial improvement, while Group III showed rapid normalization and fastest resolution of diarrhea which was clinically significant (Handekar et al., 2010; Bhatt et al., 2009; Kasaralikar et al., 2014; Syed et al., 2017).
 
Milk production and quality
 
Milk examination results (Table 5) showed stable milk yield in Group I (P = 0.547), reflecting normal variation in healthy cows. In contrast, Groups II and III exhibited highly significant increases in milk yield (P = 0.000), with Group III showing the highest production by Day 7, consistent with reports on probiotic and herbal supplementation improving lactation (Bhatt et al., 2009; Yalcin et al., 2011; Garg et al., 2025).

Table 5: Mean ± SE values of milk parameters before and after treatment.


       
Milk pH showed a mild, significant rise in Groups II (P = 0.087) and III (P = 0.041), while remaining stable in Group I (P = 0.361), indicating improved udder health. Milk protein remained largely unchanged across all groups (3.07–3.77%) (Lingathurai et al., 2009; Hossain et al., 2010), with no significant variation (P>0.05), though Group III showed a slight increase.
       
Milk fat increased significantly in Groups II (P = 0.002) and III (P = 0.000), with highest values in Group III, reflecting improved metabolic status. Similarly, SNF increased significantly only in Group III (P = 0.002), while Groups I and II showed no significant change (P = 0.547; P = 0.397), aligning with Jawale et al., (2022); Chappalwar et al., (2014) and Ali Ibrahim et al. (2012). Overall, Group III showed the most consistent improvement across all milk parameters.
       
The therapeutic efficacy of all treatment regimens was assessed based on clinical recovery, normalization of rumen function and milk parameters. Group III (nutritional supplement) showed the fastest and most effective recovery, with all cows fully recovered by days 3-5, whereas Group II (standard treatment) recovered by Days 5-7, indicating effectiveness but slower response. Overall, nutritional supplementation significantly improved rumen function, digestion and productivity, demonstrating superior efficacy in managing ruminal disorders in cattle.

CONCLUSION

Nutritional supplement used in this research, a Rumenotonic with digestive enzymes, amino acids, probiotic and herbal extract, offer an effective strategy for managing simple indigestion in lactating cows by modulating ruminal pH, enhancing rumen motility and promoting the growth and activity of beneficial microbial populations, thereby improving overall digestion and milk production. The supplement was safe and well-tolerated at recommended dose, showing no adverse effects, while also facilitating the restoration and stabilization of appetite and normal digestive functions, supporting both animal health and productivity.

ACKNOWLEDGEMENT

Authors are thankful to Associate Dean, College of Veterinary and Animal Sciences, MAFSU, Parbhani, Maharashtra, India and all staffs, PG and Ph.D. Students of Department of Veterinary Medicine for their valuable help and support during project work.
 
Funding
 
The clinical trial project was funded by M/s Advenza Global Limited Mumbai, Maharashtra, India.
 
Disclaimers
 
The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.

CONFLICT OF INTEREST

The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.

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    Full Research Article

    Assessment of The Digestive Efficacy of a Nutritional Supplement in Bovines

    M
    M.P. Sakhare1
    S
    S.P. Risbud2,*
    M
    M.F.M.F. Siddiqui1
    P
    P.C. Rathi2
    C
    C.L. Rathi2
    V
    V.K. Munde1
    A
    A.M. Chappalwar1
    A
    A.M. Syed1
    S
    S.R. Shaikh1
    1Department of Veterinary Epidemiology and Preventive Medicine College of Veterinary and Animal Sciences, Maharashtra Animal and Fishery Science Unviersity, Parbhani-431 402, Maharashtra, India.
    2Advenza Global limited (formerly known as Advanced Vital Enzymes Pvt. Ltd.), Wagle Estate, Thane(W)-400 604, Maharashtra, India.
    Cite article:- Sakhare M.P., Risbud S.P., Siddiqui M.F.M.F., Rathi P.C., Rathi C.L., Munde V.K., Chappalwar A.M., Syed A.M., Shaikh S.R. (2026). Assessment of The Digestive Efficacy of a Nutritional Supplement in Bovines . Asian Journal of Dairy and Food Research. 45: 42-50. doi: 10.18805/ajdfr.DR-2468.

    ABSTRACT

    Background: Digestive disorders in cattle impair rumen function, feed utilization and productivity. Nutritional strategies such as probiotics, enzymes and herbal extracts may enhance ruminal health. This study evaluated the efficacy of a rumenotonic bolus containing probiotics, enzymes and herbal extracts in managing ruminal disorders in bovines.

    Methods: A clinical trial was conducted on 18 lactating cows with ruminal disorders at the Department of Veterinary Medicine, Parbhani (February-June 2025). Animals were divided into three groups: Group I (healthy control), Group II (anorexia treated with standard therapy) and Group III (anorexia treated with rumenotonic bolus, 2 boli BID for 5 days). Clinical, ruminal, hematological, fecal and milk parameters were assessed on days 0, 5 and 7.

    Result: Ruminal motility, pH and fermentation parameters improved significantly in treated groups, with the greatest response in Group III. Hematological indices normalized and fecal consistency improved, with faster resolution of diarrhoea in Group III. Milk yield, fat and solids-not-fat increased significantly, while protein remained stable. Group III showed the most rapid recovery (Days 3-5), compared to slower improvement in Group II (Days 5-7). Overall, the rumenotonic supplement enhanced ruminal function and clinical recovery more effectively than standard treatment.

    INTRODUCTION

    India possesses one of the largest bovine populations globally, with 192.49 million cattle and 109.85 million buffaloes (Government of India, 2019). Maharashtra ranks fifth in cattle and seventh in buffalo population nationally. Cattle and buffaloes constitute 35.94% and 20.45% of India’s livestock population, respectively, highlighting their vital role in the rural economy and dairy sector. However, productivity is often compromised by health disorders, particularly digestive ailments, among which ruminal disorders are most prevalent.
           
    In ruminants, conditions such as anorexia, tympany and indigestion are characterized by reduced feed intake, altered ruminal pH, decreased motility and reduced protozoal counts. Simple indigestion commonly arises from sudden dietary changes, poor-quality fodder, imbalanced roughage-to-concentrate ratios, or inadequate water intake (Biswal et al., 2016). These disturbances may lead to ruminal acidosis or alkalosis, depending on fermentation patterns (Constable et al., 2017). Additionally, indigestible feed accumulation and poor feeding practices further aggravate ruminal dysfunction.
           
    The rumen, the largest and most vital digestive organ of ruminants, functions as a fermentation chamber housing a complex microbial ecosystem that maintains physiological homeostasis. The rumen is responsible for fermenting fibrous feeds into volatile fatty acids, the primary energy source for ruminants (Moran, 2005; Smith and Jones, 2009). Disruption of this microbial balance adversely affects nutrient utilization and predisposes animals to metabolic and digestive disorders such as acidosis, diarrhea, ketosis, mastitis and laminitis (Asrat et al., 2015; Pan, 2016; Zhao et al., 2023). Maintaining ruminal health is therefore essential for optimal productivity (Beauchemin, 2002).
           
    The rationale for this study is based on the need for effective nutritional interventions that can restore ruminal microbial balance, improve digestion and enhance recovery from digestive disorders. Probiotics and exogenous enzymes have gained importance as nutritional interventions to restore ruminal function. These supplements enhance microbial balance, improve digestion and increase feed efficiency (Anadon et al., 2006; Uyeno et al., 2015). Enzymes, amino acids and herbal additives further support nutrient utilization and recovery during anorexia (Arowolo et al., 2018; Mazinani et al., 2022; Chaudhary et al., 2021; Ramdani et al., 2025).
           
    Considering the high incidence and economic impact of ruminal disorders, the present study was undertaken with an objective to evaluate the efficacy of a multi-component nutritional supplement (bolus) containing probiotics, enzymes, amino acids and herbal extracts in improving ruminal function and managing ruminal disorders in bovines.

    MATERIALS AND METHODS

    Study area
     
    The present clinical trial was conducted on 18 lactating cows presented to the Department of Veterinary Medicine, Veterinary Clinical Complex (VCC), Instructional Livestock Farm Complex (ILFC), College of Veterinary and Animal Sciences, Maharashtra Animal and Fishery Sciences University (MAFSU), Parbhani and nearby veterinary dispensaries in Parbhani district during February to June 2025. The study evaluated the effect of a nutritional supplement on ruminal disorders in lactating cows, with emphasis on rumen function, milk yield and milk composition.
     
    Test product
     
    The investigational product is a Rumenotonic with digestive enzymes, amino acids, probiotic and herbal extract supplement containing Amylase, Cellulase, Xylanase - Lysine-DL, Methionine, Histidine, Leucine, Valine, Threonine, Lactobacillus acidophilus, Bacillus coagulans, Saccharomyces boulardii, Aspergillus oryzae, ginger powder and soyabean meal, supplied by Advenza Global Limited in the form of a bolus.
     
    Study design
     
    Lactating cows diagnosed with ruminal disorders, including simple indigestion, acidic indigestion, alkaline indigestion and anorexia of unknown origin, irrespective of age and breed, were selected based on detailed history and thorough clinical examination. Observations were recorded on day 0 (before treatment), day 5 and day 7 (after treatment).
           
    Eighteen cattle were divided into three groups (n = 6 each): Group I comprised healthy animals maintained under identical feeding and management conditions (control); Group II included clinical cases of anorexia of unknown origin treated with a standard protocol and Group III consisted of similar anorexic cases treated with a nutritional supplement administered as two boli twice daily for five days.
     
    Sample size
     
    The sample size was determined based on feasibility, ethical considerations and availability of animals with similar clinical conditions. A total of 18 animals were included and divided into three groups (Group I, II and III; n = 6 each). This size was considered sufficient to detect biologically meaningful differences while minimizing animal use, consistent with previous studies (Charan and Kantharia, 2013).
     
    Clinical and physiological parameters
     
    A detailed anamnesis and clinical examination were conducted for all animals. Heart rate was recorded by auscultation over the left thoracic region (3rd-5th intercostal space) and expressed as beats per minute. Respiratory rate was measured by observing nostril movement and thoracic auscultation, expressed as breaths per minute. Rectal temperature was recorded using a digital thermometer (°F). Rumen motility was assessed by palpation and auscultation of the left paralumbar fossa and expressed as contractions per two minutes. Bloat and anorexia were graded using standardized 0-3 scoring scales.
     
    Rumen fluid collection and examination
     
    Rumen fluid was collected aseptically from the left paralumbar fossa using an 18G, 6-inch spinal needle and extraction pump (Fig 1 and Fig 2). Physical characteristics (colour, odour, consistency) were visually assessed (Fig 3 and Fig 4). pH was measured using indicator paper (HiMedia Laboratories) and categorized as mild (5.5-6.5), moderate (4.5-5.5) and severe (4.0-4.5) acidity (Fig 5 and Fig 6). Total protozoal count was estimated using the method of Moir (1951) and expressed as ×105 /ml.

    Fig 1: Collection of ruminal fluid by stomach tube.



    Fig 2: Collection of ruminal fluid from left paralumbar fossa.



    Fig 3: Brownish green rumen fluid.



    Fig 4: Yellowish green rumen fluid.



    Fig 5: Normal rumen fluid pH hematological, fecal and milk analysis.



    Fig 6: Alkaline pH of rumen fluid.


           
    Blood samples were collected on day 0, 5 and 7 into EDTA (3.5%) vacutainers and analyzed using an automatic hematology analyzer (Spincell 5). Fecal examination assessed colour, odour, consistency and diarrhea. Milk yield and composition (pH, fat, protein, Solid -Non-Fat) were recorded and analyzed using a LactoScan analyzer (Fig 7 and Fig 8).

    Fig 7: Automatic milk analyzer.



    Fig 8: Estimation of milk samples.


     
    Statistical analysis
     
    Data obtained were subjected to statistical analysis following the methods described by Snedecor and Cochran (1994). Analysis was performed using Completely Randomized Design (CRD) with SPSS version 16. Results were expressed as mean ± standard error (SE).

    RESULTS AND DISCUSSION

    The present study evaluated efficacy of a multi-component nutritional supplement on ruminal disorders in lactating cows by assessing clinical, physiological, ruminal, haematological, fecal and milk production parameters. Observations recorded at different time intervals (day 0, 5 and 7) provided insights into the progression and recovery pattern in treated animals compared to controls. The findings are discussed in relation to improvements in rumen function, microbial activity and overall health status.
     
    Physiological and clinical examination
     
    The physiological and clinical parameters in cows affected with simple indigestion showed anorexia, mild depression, reduced feces which were initially dry and and later became softer, voluminous and malodorous consistent with Kasaralikar et al., (2014); Syed et al., (2017) and Constable (2017). The heart rate, respiration rate and rectal temperature remained within normal limits and comparable to controls as presented in Table 1. Post-treatment, no significant changes (p<0.05) occurred except a slight heart rate increase in Group III (p = 0.048), remaining within normal range, similar to previous reports (Kasaralikar et al., 2014; Syed et al., 2017; Constable et al., 2017).

    Table 1: The Physiological parameters of animals before and after treatment.


           
    Ruminal motility (Table 1) showed non-significant variation in Group I, while indigestion-affected cows in Groups II and III exhibited reduced baseline motility on Day 0, consistent with earlier reports (Walia et al., 2011; Kasaralikar et al., 2014). Group II showed significant improvement during treatment, whereas Group III demonstrated a highly significant progressive increase from 1.33±0.21 to 2.83±0.17 (p = 0.000), with Day 5 and Day 7 values significantly higher than baseline. This restoration approached normal ruminal motility (~3 contractions/2 minutes) as reported by Chakrabarti (2014) and Constable et al., (2017). Among all groups, Group III showed the greatest improvement.
           
    In Group I, the left paralumbar fossa was normal, whereas mild distension was observed in a few cows (n = 5) in Groups II and III and the remaining (n = 13) showed a firm, doughy rumen. These findings agree with Constable et al., (2017). Post-treatment, all cows in Groups II and III showed normal paralumbar fossa, indicating resolution of distension, consistent with Walia et al., (2011) and Hussain and Uppal (2012).
           
    Appetite remained normal in Group I, while reduced appetite was observed in all affected cows before treatment, as also reported by Constable et al., (2017). Post-treatment, normal appetite was restored in all cows of Groups II and III, indicating effective clinical recovery.
     
    Rumen fluid examination
     
    Physical examination
     
    Healthy cows (Group I) exhibited greenish to yellowish-brown rumen fluid with aromatic odour and viscous-to-thick consistency (Handekar et al., 2010; Kasaralikar et al., 2014). In simple indigestion, it changed to yellowish-green to light brown with occasional ammoniacal odour and watery to semi-solid consistency (Handekar et al., 2010). Post-treatment in Groups II and III, parameters normalized, indicating restoration of rumen function (Waghmare et al., 2009). Observations are presented in Table 2.

    Table 2: The physical rumen fluid examination parameters before and after treatment.


     
    Physicochemical and microbiological findings
     
    The mean rumen liquor pH remained within the normal range in control cows (Chakrabarti, 2014; Constable et al., 2017), while significantly higher values (p<0.01) were observed in Groups II and III before treatment, indicating ruminal alkalinization (Biswal et al., 2016). Post-treatment, pH declined significantly in Groups II and III, reaching near-normal levels by Day 5 and Day 7.
           
    Protozoal counts in Group I showed a mild, non-significant increase, while Groups II and III had significantly lower counts pre-treatment with marked post-therapy improvement, especially in Group III (p<0.05), consistent with Mohan et al., (2015). This recovery is closely linked to rumen pH, as protozoa are highly pH-sensitive (Granja-Salcedo et al., 2016) and ruminal acidosis reduces microbial balance through excess volatile fatty acids (TVFA) and lactic acid formation (Biswal et al., 2016). Protozoa play a key role in fermenting carbohydrates into acetic and butyric acids (Chakrabarti, 2014), so their restoration indicates improved fermentation efficiency (Petrovski, 2016).
           
    TVFA levels remained normal in Group I (6-12 mEq/dl; Chakrabarti, 2006), while Groups II and III showed significant changes (p<0.01), with Group III exhibiting increased levels (Bhatt et al., 2009). Ammonia nitrogen remained stable in Group I but improved significantly in Group III, indicating enhanced microbial protein synthesis (Shen et al., 2023). Mean values are presented in Table 3.

    Table 3: The rumen fluid examination parameters before and after treatment.


     
    Hematological findings
     
    The results of hematological examination of all groups are presented in Table 4. Haemoglobin and total erythrocyte count increased significantly in Groups II and III, while total leukocyte count decreased significantly, indicating recovery. Other leukocyte parameters remained mostly unchanged. These findings align with earlier studies (Kasaralikar et al., 2014; Syed et al., 2017).

    Table 4: Hematological parameters of all groups before and after treatment.


     
    Fecal examination
     
    Fecal examination showed normal greenish-brown, semisolid feces in Group I (Biswal et al., 2016). In Groups II and III, feces were yellowish-brown, loose and foul-smelling, indicating impaired fermentation (Biswal et al., 2016; Petrovski, 2016). Post-treatment, Group II partial improvement, while Group III showed rapid normalization and fastest resolution of diarrhea which was clinically significant (Handekar et al., 2010; Bhatt et al., 2009; Kasaralikar et al., 2014; Syed et al., 2017).
     
    Milk production and quality
     
    Milk examination results (Table 5) showed stable milk yield in Group I (P = 0.547), reflecting normal variation in healthy cows. In contrast, Groups II and III exhibited highly significant increases in milk yield (P = 0.000), with Group III showing the highest production by Day 7, consistent with reports on probiotic and herbal supplementation improving lactation (Bhatt et al., 2009; Yalcin et al., 2011; Garg et al., 2025).

    Table 5: Mean ± SE values of milk parameters before and after treatment.


           
    Milk pH showed a mild, significant rise in Groups II (P = 0.087) and III (P = 0.041), while remaining stable in Group I (P = 0.361), indicating improved udder health. Milk protein remained largely unchanged across all groups (3.07–3.77%) (Lingathurai et al., 2009; Hossain et al., 2010), with no significant variation (P>0.05), though Group III showed a slight increase.
           
    Milk fat increased significantly in Groups II (P = 0.002) and III (P = 0.000), with highest values in Group III, reflecting improved metabolic status. Similarly, SNF increased significantly only in Group III (P = 0.002), while Groups I and II showed no significant change (P = 0.547; P = 0.397), aligning with Jawale et al., (2022); Chappalwar et al., (2014) and Ali Ibrahim et al. (2012). Overall, Group III showed the most consistent improvement across all milk parameters.
           
    The therapeutic efficacy of all treatment regimens was assessed based on clinical recovery, normalization of rumen function and milk parameters. Group III (nutritional supplement) showed the fastest and most effective recovery, with all cows fully recovered by days 3-5, whereas Group II (standard treatment) recovered by Days 5-7, indicating effectiveness but slower response. Overall, nutritional supplementation significantly improved rumen function, digestion and productivity, demonstrating superior efficacy in managing ruminal disorders in cattle.

    CONCLUSION

    Nutritional supplement used in this research, a Rumenotonic with digestive enzymes, amino acids, probiotic and herbal extract, offer an effective strategy for managing simple indigestion in lactating cows by modulating ruminal pH, enhancing rumen motility and promoting the growth and activity of beneficial microbial populations, thereby improving overall digestion and milk production. The supplement was safe and well-tolerated at recommended dose, showing no adverse effects, while also facilitating the restoration and stabilization of appetite and normal digestive functions, supporting both animal health and productivity.

    ACKNOWLEDGEMENT

    Authors are thankful to Associate Dean, College of Veterinary and Animal Sciences, MAFSU, Parbhani, Maharashtra, India and all staffs, PG and Ph.D. Students of Department of Veterinary Medicine for their valuable help and support during project work.
     
    Funding
     
    The clinical trial project was funded by M/s Advenza Global Limited Mumbai, Maharashtra, India.
     
    Disclaimers
     
    The views and conclusions expressed in this article are solely those of the authors and do not necessarily represent the views of their affiliated institutions. The authors are responsible for the accuracy and completeness of the information provided, but do not accept any liability for any direct or indirect losses resulting from the use of this content.

    CONFLICT OF INTEREST

    The authors declare that there are no conflicts of interest regarding the publication of this article. No funding or sponsorship influenced the design of the study, data collection, analysis, decision to publish, or preparation of the manuscript.

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