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. 2025 Feb 5;15:4442. doi: 10.1038/s41598-025-89026-9

Effect of supplementation of phytoadditive mixtures on live weight, blood indices, hormonal profile and reproductive performance in Black Bengal goats

Amit Kumar Singh 1,2,✉, Champak Bhakat 1, Upali Kisku 1, Muthupalani Karunakaran 1, Tapas Kumar Dutta 1
PMCID: PMC11799428  PMID: 39910342

Abstract

The study was conducted to evaluate the effect of phytoadditive mixtures containing equal proportions of Curcuma longa & Allium sativum (PAM-1) and Curcuma longa & Ocimum sanctum (PAM-2) on live weight, blood parameters, hormonal profile and reproductive performance in Black Bengal goats. A total of 21 post weaned female kids of 3–4 months age and similar body weights maintained at Goat Research Unit of Eastern Regional Station (ERS), National Dairy Research Institute (NDRI), Kalyani, India during the year 2021–2022 were randomly distributed in three groups namely, Control, PAM-1 and PAM-2 for 10 months. Blood parameters i.e. glucose, urea, total protein, albumin, globulin, ALT, AST, ALP, total lipids, cholesterol, and Triglycerides, WBC, RBC, Hb and PCV %; NEFA levels and hormones i.e. prolactin, cortisol, T3 and T4 were estimated in all three experimental groups. Higher (P < 0.001) live weights were investigated in supplemented goats. Blood parameters including glucose, urea, total protein, albumin, globulin, ALT, AST, ALP, WBC, RBC, Hb and PCV% were similar (P > 0.05) in all groups. Lower (P < 0.001) blood lipids were found in supplemented groups. Higher Prolactin, T3 and T4 levels and lower Cortisol levels were reported in PAM-1 and PAM-2 groups (P < 0.001) than control group. Age at first estrus, first conception, first kidding and post kidding first estrus days were lower (P < 0.001) in supplemented groups. Therefore, it was concluded that supplementation of PAM-1 and PAM-2 improved reproductive performance, hormonal profile and lipid profile through improved live weights in Black Bengal goats.

Keywords: Additive supplementation, Allium sativum, Blood indices, Body growth, Curcuma longa, Goats, Hormones, Ocimum sanctum, Phytoadditive feeding, Reproduction

Subject terms: Biochemistry, Biotechnology, Zoology

Introduction

Black Bengal goats contribute to the highest number of goats in India1. They are predominant goat breed in lower Gangetic region2. Black Bengal goats are considered as prolific breed but, under poor nutritional management they do not perform well3. Earlier, antibiotics were utilized in animal feed for improving growth and reproductive performance of animals4,5. However, due to strict government rules and awareness in consumers, the use of antibiotics in animal feed for augmenting their reproductive performances has declined drastically as there is potential risk of antibiotic resistance and environmental degradation with the use of antibiotics in animal feed6,7. This has increased the interest in use of alternative feed additives in animal nutrition8,9. Recently, phytoadditives have emerged as one of the potential alternative feed additives in animal nutrition4,10.

Phytoadditives are derived from different parts of plants containing plant secondary metabolites for their natural defense against invading pathogens11,12. Earlier studies have indicated that use of phytoadditives may enhance nutrient utilization, growth performance, lipid profile and reproductive performance7,13,14 among other parameters in farm animals due to their antioxidant and hormone regulating properties15–17. Phytoadditives such as Curcuma longa, Allium sativum and Ocimum sanctum possess anti-oxidant, anti-microbial properties, improves nutrient utilization due to the presence of plant secondary metabolites such as total phenolics, tannins, saponins, flavonoids and alkaloids18–20. Phytochemicals namely, Curcumin, Allicin and Eugenol are major bio-active compounds in Curcuma longa, Allium sativum and Ocimum sanctum11,18,21. Phytoadditives like Curcuma longa, Allium sativum and Ocimum sanctum are economically and abundantly available in Indian conditions especially in West Bengal state of India22,23.

To the current knowledge, no study has been conducted to understand the effect of Phyto additive mixtures on live weight, hormonal profile and reproductive performance of Black Bengal goats in addition to blood indices. We hypothesized that phytoadditive mixtures containing equal proportions of Curcuma longa & Allium sativum and Curcuma longa & Ocimum sanctum may affect blood indices, hormonal profile and reproductive performances by improving live weight in Black Bengal goats. Therefore, the study was framed to understand the effect of Phytoadditive mixtures containing equal proportions of Curcuma longa & Allium sativum (PAM 1) and Curcuma longa & Ocimum sanctum (PAM 2) on live weight, blood indices, hormones and reproductive performance of Black Bengal goats.

Materials and methods

Location of study

The experiment was carried out at Goat Research Unit of Eastern Regional Station (ERS), National Dairy Research Institute (NDRI), Kalyani, India during the year 2021–2022. Every activity of this goat unit is supervised by scientists as per the standard farm management practices approved by Ethical Committee of the Institute (registration: 19/P/LP/09). Latitude and longitude of Kalyani being 22.975084, and 88.434509. Typically, the year-round temperature ranges from 12.78 °C to 38.89 °C with an average of 30.26 °C and receives 96.3 millimetres of rainfall annually. This area comes under alluvial regions of tropical lower Gangetic region. Kalyani lies 11 m above mean sea level. This experiment was conducted from first week of January, 2021 to May, 2022.

Experimental animals and management practices

A total of 21 post weaned female Black Bengal goat kids of 3–4 months age and body weights (5.07 ± 0.52 kg) were randomly distributed into 3 comparable groups each having 7 animals. Individual feeding of kids was practiced in this study. All the kids received similar housing and other management practices [Beuro of Indian Standards, 2733: 1985]. Ad libitum feeding was provided to the kids with 10 kg capacity buckets however each time it was measured and recorded with the help of electronic weighing scale with a least count of 5.0 g. Seasonal green and dry fodders and fresh clean water were available to each kid for 24 h. Feeding management was done as per standard feeding practices recommended by NRC, 200724. Calculated cost of concentrate mixture and roughage mixture was Rs. 32.0/kg and Rs. 2.0/kg, respectively. Natural breeding was performed with Black Bengal bucks of Goat Research Unit at ERS of NDRI. Same breeding practice was adopted for every goat after confirming oestrus symptoms. Record of individual kid was utilized for age related determinations.

Estimation of plant secondary metabolites in selected herbs

A sample of 10 g powders of three phytoadditives were taken in separate conical flasks and mixed with 100 ml of CH3OH and the conical flasks were plugged with cotton. The obtained solution was kept under continuous stirring for 24 h. After this, the supernatant was filtered with filter paper 41 and solvent was evaporated using vacuum evaporator (operated at 42 –45 °C) to obtain crude extract25. This crude extract was stored in air tight containers for storage under refrigerated conditions (5 °C) for further estimations. Plant secondary metabolites such as flavonoids26, saponins27, alkaloids28 and total phenolics, present [Table 1] in three herbs were determined as per the methods adopted by Soni and Sosa25 with the help of spectrophotometer. Tannic acid (standard phenolic compound) and Folin- Ciocalteu were used as reagent and the absorbance of final blue colour solution was measured at 760 nm under spectrophotometer for determination of total phenolics compounds. All the samples were analyzed in triplicates on every two months during growth trial in this study.

Table 1.

Nutrient composition and plant secondary metabolites in feeds and phytoadditives in diets of Black Bengal goats.

Composition Concentrate mixture Roughage mixture Curcuma longa Allium sativum Ocimum sanctum
DM % 90.73 ± 0.14 24.92 ± 0.19 22.28 ± 0.59 33.51 ± 0.02 13.21 ± 0.02
Moisture % 9.27 ± 0.14 75.08 ± 0.19 77.72 ± 0.59 66.49 ± 0.02 86.79 ± 0.02
CP %DM 20.64 ± 0.13 12.88 ± 0.24 11.17 ± 0.02 12.49 ± 0.01 18.21 ± 0.03
CF %DM 11.61 ± 0.15 23.79 ± 0.22 4.57 ± 0.01 2.06 ± 0.03 9.04 ± 0.01
EE %DM 3.42 ± 0.09 2.85 ± 0.09 12.39 ± 0.01 2.39 ± 0.02 3.33 ± 0.01
Ash %DM 3.3 ± 0.05 9.93 ± 0.10 7.79 ± 0.01 1.75 ± 0.02 13.15 ± 0.02
Plant secondary metabolites (mg/100 g)
Total phenolics - - 167.85 ± 0.08 143.19 ± 0.14 176.24 ± 0.21
Flavonoids - - 3.78 ± 0.01 5.48 ± 0.01 97.15 ± 0.26
Saponins - - 2.29 ± 0.01 0.80 ± 0.01 4.79 ± 0.01
Alkaloids - - 6.60 ± 0.01 4.16 ± 0.01 6.42 ± 0.01
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Table 2.

Nutrient utilization by Black Bengal goats.

Parameters Control PAM-1 PAM-2 SEM P value
Intake parameters
TDMI (g/day/goat) 365.73a 399.11b 398.60b 10.03 < 0.001
TDMI of %BW 3.58a 3.57a 3.58a 0.01 0.213
Nutrient digestibility (%)
DM 62.42a 63.92b 63.77b 0.19 < 0.001
CP 72.23a 73.99b 74.24b 0.11 < 0.001
EE 72.93a 74.07b 74.05b 0.14 < 0.001
ADF 46.23 46.15 46.06 0.34 0.940
NDF 60.68 60.78 60.91 0.25 0.803
Hemi cellulose 67.63a 69.05b 69.14b 0.16 < 0.001
OM 65.95a 69.29b 69.20b 0.14 < 0.001
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Means bearing different superscripts differ significantly (P < 0.05) across different columns.

Table 3.

LSQ means of blood indices of Black Bengal goats supplemented with phytoadditive mixtures containing Curcuma longa, Allium sativum and Ocimum sanctum.

Parameters Control PAM-1 PAM2 SEM P value
Blood metabolites
Glucose (mg/dL) 52.50 52.58 52.53 0.03 0.178
Total Protein (mg/dL) 7.04 7.06 7.06 0.01 0.056
Albumin (mg/dL) 3.55 3.56 3.55 0.01 0.140
Globulin (mg/dL) 3.49 3.51 3.50 0.01 0.097
Albumin: Globulin 1.02 1.01 1.01 0.01 0.559
Total urea (mg/dL) 12.76 12.76 12.77 0.01 0.087
Blood enzymes
ALT (IU/L) 33.99 33.97 34.03 0.06 0.782
AST (IU/L) 63.56 63.47 63.50 0.06 0.610
ALP (IU/L) 167.42 167.53 167.37 0.05 0.113
Blood haematology
WBC (106/mm3) 7.77 7.78 7.79 0.01 0.177
RBC (106/mm3) 8.00 8.01 8.01 0.01 0.454
Hb (g %) 10.92 10.92 10.94 0.01 0.098
PCV (%) 27.67 27.69 27.63 0.03 0.396
Blood lipids
Total lipids (mmol/L) 7.90a 7.39b 7.43b 0.02 < 0.001
Cholesterol (mg/dL) 2.53a 2.05b 2.04b 0.02 < 0.001
Triglycerides (mg/dL) 2.22a 1.97b 1.96b 0.01 < 0.001
NEFA (mmol/L) 0.278a 0.232b 0.235b 0.001 < 0.001
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Means bearing different superscripts differ significantly (P < 0.05) across different columns.

Table 4.

Reproductive performance of Black Bengal goats with phytoadditive mixtures containing Curcuma longa, Allium sativum and Ocimum sanctum.

Parameters (days) Control PAM-1 PAM-2 SEM P value
Age at first estrus 214.86a 193.86b 198.14b 2.45 < 0.001
Age at first conception 269.86a 234.14b 231.43b 3.77 < 0.001
Age at first kidding 418.29a 383.29b 380.14b 6.79 < 0.001
Gestation length 148.43 149.14 148.71 2.38 0.422
Post kidding first estrus 38.57a 31.71b 33.29b 1.55 < 0.001
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Means bearing different superscripts differ significantly (P < 0.05) across different columns.

Preparation and feeding of phytoadditive mixtures

Two different phytoadditive mixtures were prepared for this study. First phytoadditive mixture comprised equal proportions of Curcuma longa & Allium sativum (PAM 1) and second mixture contained equal proportions of Curcuma longa & Ocimum sanctum (PAM 2). Good quality Curcuma longa rhizomes (fully grown and freshly harvested), Allium sativum (cloves obtained after peeling outer skin) and Ocimum sanctum (middle aged) leaves were procured from local market during December, 2020 and powdered as per the standard powdering process followed by Oderinwale et al.18. Per kg cost (on fresh basis) of Curcuma longa & Allium sativum and Ocimum sanctum were Rs. 50.0, Rs. 200.0 and Rs. 50.0, respectively. After procuring, the ingredients were cleaned and sundried followed by thin slicing and oven dried. Hot air oven was operated at 60 °C for 24 h. Dried ingredients were separately grinded finely with the help of a mechanical grinder (750 W Borosil ARO grinder, Borosil limited, Bandra East, Mumbai − 500 051, Maharashtra, India) to pass through a screen of 1 mm. After final grinding, the powders of Curcuma longa, Allium sativum and Ocimum sanctum were obtained and two phytoadditive mixtures were obtained after thorough mixing followed by their storage in air tight and sterilized vessels until further use. Phytoadditive mixture (PAM 1) containing Curcuma longa and Allium sativum (1:1 ratio) was provided to PAM 1 group kids (15 g/kg of DM) in concentrate mixture and phytoadditive mixture (PAM 2) containing Curcuma longa and Ocimum sanctum (1:1 ratio) was provided to PAM 2 group kids (15 g/kg of DM) in concentrate mixture. Before taking observations, all kids were provided with mixture 14 days as adaptation period.

Chemical composition of feeds and phytoadditives

Refusal fodder was collected and measured daily in the morning 7:00 AM to 8:00 AM before morning feeding. Fodder and phytoadditives samples were dried in hot air oven at 60 °C for 24 h. Dried samples were powdered with the help of a mechanical grinder to pass through a screen of 1 mm. The powdered samples were analyzed for crude protein [29, Method 990.03], ether extract [29, Method 920.39] and ash content [29, Method 942.05] in the feed on DM basis. The NDF and ADF contents were analyzed using method of Van Soest et al.30. Table1 represents the nutritional composition of concentrate mixture, roughage mixture, Curcuma longa (turmeric), Allium sativum (garlic) and Ocimum sanctum (tulsi) powder used in the experimental diets of Black Bengal kids.

Nutrient utilization and live weight of goats

Daily feed intake (FI) and fortnightly live weight of Black Bengal kids were accessed at 8:00 AM to 9:00 AM before morning feeding. An electronic weighing balance (KERN CH50K50, GmbH, D-72336 Ballingen, Germany) was used for measurements of feed intake and live weights of animals. A representative sample of residual roughage for each goat was collected in pre marked plastic zipper bags followed by drying at 60 °C for 24 h in hot air oven for determination of actual DMI by the kids. Digestibility trial of goats for 7 days was performed after the end of growth trial. Faecal samples were collected in pre marked plastic zipper bags for each goat followed by hot air oven drying at 50 °C to reach on constant DM. Dried fecal samples were grounded in mechanical grinder. The grounded samples were analyzed for crude protein [29, Method 990.03], ether extract [29, Method 920.39] and ash content [29, Method 942.05] in the feed on DM basis. The NDF and ADF contents were analyzed using method of Van Soest et al.30.

Analysis of blood samples

Blood samples were collected in a sterile 10 ml calibrated tube by jugular vein puncture of individual goat on monthly intervals over 13 months of experiment before morning feeding by the experience persons without disturbing other goats and the samples were stored under − 20 °C until further estimations. Blood parameters such as glucose, urea, total protein, albumin, globulin, Alanine transaminase (ALT), Aspartate aminotransferase (AST), Alkaline phosphatise (ALP), Total lipids, cholesterol, and Triglycerides, White blood cells (WBC), Red blood cells (RBC), Haemoglobin (Hb), Packed cell volume (PCV %) (ARKRAY Healthcare Pvt. Ltd., Bardhman, West Bengal, India) were assessed though spectrophotometer.

Non-esterified fatty acids (NEFA) levels (ImmunoTag goat NEFA kit, Code: ITE110080, G-Bioscience) and Hormones including Prolactin (ImmunoTag goat PRL kit, Code: ITE110039, G-Bioscience), Cortisol (ImmunoTag goat COR kit, Code: ITE110021, G-Bioscience), Triiodothyronine (T3) (ImmunoTag goat T3 kit, Code: ITU00020A, G-Bioscience) and Thyroxine (T4) (ImmunoTag goat T4 kit, Code: ITE110067, G-Bioscience) concentrations were estimated on monthly intervals of each goat through commercial ELISA kits of G-Bioscience (G-Bioscience 9800, Page Avenue, St. Louis, MO, USA) using a micro plate reader (BioTek EPOCH2NS).

Statistical analysis

Randomized complete research design was followed in this study. All the collected data were subjected to statistical analysis using univariate general linear model procedure of statistical tool SPSS 26.0 (IBM Co.) with treatments and time as fixed factors. MS Excel 2007 was used for data tabulation and graphical representation. Duncans’ multiple range test (DMRT) was used to determine significant differences among different mean values. Means were compared at different significant levels (P < 0.05 and P < 0.01).

Results

Nutrient composition and plant secondary metabolites

The nutrient composition and plant secondary metabolites in feeds and phytoadditives in diets of Black Bengal goats have been presented in table 1. Dry matter (%) in concentrate mixture, roughage mixture, Curcuma longa, Allium sativum and Ocimum sanctum were 90.73, 24.92, 22.28, 33.51 and 13.21, respectively. While, Crude protein content (% DM) were 20.64, 12.88, 11.17, 12.49 and 18.21, respectively. Highest content of total phenolics (176.24 mg/100g), flavonoids (97.15 mg/100g), saponins (4.79 mg/100g) was found in Ocimum sanctum, whereas, highest alkaloid concentration (6.60 mg/100g) was reported in Curcuma longa among all phytoadditives

Nutrient utilization and live weight changes

Improved total dry matter intake was investigated in treatment (PAM-1: 399.11 g/goat/day, PAM-2: 398.60 g/goat/day) than control group (365.73 g/goat/day) [Table 2] and the difference between the groups were statistically significant (P < 0.001). An additional cost of feeding of Rs. 524.0 in PAM-1 and Rs. 410.0 in PAM-2 was incurred in this experiment due to additional feeding cost as a result of increase in DMI and phyto additive supplementation than control group (1 USD = 85.74 INR). Higher live weights of supplemented goats (PAM-1: 17.51 kg/goat, PAM-2: 17.48 kg/goat) than control (15.21 kg/goat) goats were observed in this study [Fig. 1]. The values differed significantly (P < 0.01) between the groups. Digestibility of ADF and NDF were similar in all the groups (P > 0.05). Higher (P < 0.001) DM, CP, EE, hemicellulose and OM digestibility were observed in treatment groups PAM-1 and PAM-2 than control group goats.

Blood parameters

Blood parameters [Table 3] including glucose (P = 0.178), total protein (P = 0.056), albumin (P = 0.140), globulin (P = 0.097), albumin: globulin (P = 0.559), total urea (P = 0.087), ALT (P = 0.782), AST (P = 0.610), ALP (P = 0.113), WBC (P = 0.177), RBC (P = 0.154), Hb (P = 0.098) and PCV (P = 0.396) showed a non-significant difference among all groups.

The results showed [Table 3] that there was significant effect (P < 0.001) of phytoadditive mixtures on blood lipid profile of treatment PAM-1 and PAM-2 than control group goats. Higher total blood lipids were recorded in control (7.90 mmol/L) than PAM-1 (7.39 mmol/L) and PAM-2 (7.43 mmol/L) groups. Similarly, Cholesterol levels were reported highest (value) and differed statistically (P < 0.001) in control (2.53 mg/dL) than PAM-1 (2.05 mg/dL) and PAM-2 (2.04 mg/dL) groups. In addition to this, highest and significantly differed (P < 0.001) concentrations of triglycerides were observed in control (2.22 mg/dL) than PAM-1 (1.97 mg/dL) and PAM-2 (1.96 mg/dL) groups. However, blood lipids in PAM-1 and PAM-2 groups were similar (P > 0.05). As shown [Fig. 2], overall concentrations of NEFA were lower but significantly different (P < 0.001) in PAM-1 (0.232 mmol/L) and PAM-2 (0.235 mmol/L) groups than control (0.278 mmol/L) group goats. There was steeper decline in concentrations of NEFA (mmol/L) in PAM-1 and PAM-2 groups than control group goats after showing their peak levels.

Fig. 1.

Fig. 1

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Live weight changes in Black Bengal goats supplemented with phytoadditive mixtures (P<0.01). Note: PAM-1 (goats supplemented with 15g/kg DM Curcuma longa & Allium sativum in 1:1), PAM-2 (goats supplemented with 15g/kg DM Curcuma longa & Ocimum sanctum in 1:1).

Fig. 2.

Fig. 2

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NEFA (mmol/L) levels in Black Bengal goats supplemented with phytoadditive mixtures (P<0.01). Note: PAM-1 (goats supplemented with 15g/kg DM Curcuma longa & Allium sativum in 1:1), PAM-2 (goats supplemented with 15g/kg DM Curcuma longa & Ocimum sanctum in 1:1).

Fig. 3.

Fig. 3

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Prolactin (ng/ml) levels in Black Bengal goats supplemented with phytoadditive mixtures (P < 0.01). Note: PAM-1 (goats supplemented with 15 g/kg DM Curcuma longa & Allium sativum in 1:1), PAM-2 (goats supplemented with 15 g/kg DM Curcuma longa & Ocimum sanctum in 1:1).

Fig. 4.

Fig. 4

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Cortisol (ng/ml) levels in Black Bengal goats supplemented with phytoadditive mixtures (P<0.01). Note: PAM-1 (goats supplemented with 15g/kg DM Curcuma longa & Allium sativum in 1:1), PAM-2 (goats supplemented with 15g/kg DM Curcuma longa & Ocimum sanctum in 1:1)

Fig. 5.

Fig. 5

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T3 (ng/ml) levels in Black Bengal goats supplemented with phytoadditive mixtures (P < 0.01). Note: PAM-1 (goats supplemented with 15 g/kg DM Curcuma longa & Allium sativum in 1:1), PAM-2 (goats supplemented with 15 g/kg DM Curcuma longa & Ocimum sanctum in 1:1).

Fig. 6.

Fig. 6

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T4 (ng/ml) levels in Black Bengal goats supplemented with phytoadditive mixtures (P < 0.01). Note: PAM-1 (goats supplemented with 15 g/kg DM Curcuma longa & Allium sativum in 1:1), PAM-2 (goats supplemented with 15 g/kg DM Curcuma longa & Ocimum sanctum in 1:1).

Hormonal profile

Results indicated a significant (P< 0.001) effect of phytoadditive mixtures on hormonal profile of goats in treatment group goats. Prolactin levels (ng/ml) of goats under different groups have been shown in Fig3. During first two months of the experiment, prolactin levels in all group were similar (P > 0.05) but, significant difference in prolactin levels were observed in control and treatment groups from 3rd month of study. Highest levels were marked in PAM-1 (71.88 ng/ml) and PAM-2 (75.13 ng/ml) groups around 10th month (kidding month of treatment groups) of the experiment. While, highest prolactin levels in control (69.93 ng/ml) group was observed around 11th month (kidding month of control group) of the experiment. Treatment groups PAM-1 (15.00 ng/ml) and PAM-2 (15.16 ng/ml) had higher (P < 0.001) overall prolactin levels than control (12.50 ng/ml) group goats.

Figure4 depicts the levels of cortisol (ng/ml) in goats of different groups. In first month of the experiment, cortisol levels were similar (ranged from 4.93 ng/ml to 5.78 ng/ml) in all groups. However, lower levels of cortisol (P < 0.001) was reported in goats of treatment group than control group during 2nd month of the experiment. Highest levels of cortisol in treatment (PAM-1: 23.63 ng/ml; PAM-2: 25.38 ng/ml) groups reached around 10th month of the experiment. While, highest cortisol levels in control (26.64 ng/ml) group was observed around 11th month of the experiment. Treatment groups PAM-1 (8.17 ng/ml) and PAM-2 (8.13 ng/ml) had lower (P < 0.001) overall cortisol levels than control (8.81 ng/ml) group goats.

Levels of T3(ng/ml) in goats of different groups has been represented in Fig5. During first two months of the experiment, T3 levels in all group were similar but, there was significant difference (P < 0.01) in control and treatment groups from 3rd month of study. Highest levels were marked in PAM-1 (2.29 ng/ml) and PAM-2 (2.24 ng/ml) groups around 9th month of the experiment. While, highest T3 levels in control (2.18 ng/ml) group was observed around 10th month of the experiment. Treatment groups i.e. PAM-1 (1.88 ng/ml) and PAM-2 (1.87 ng/ml) had higher overall T3 levels than control (1.79 ng/ml) group goats and there were statistically difference (P < 0.001) between those groups.

Levels of T4 (ng/ml) in goats of different groups has been represented in Fig6. During first two months of the experiment, T4 levels in all group were similar (P > 0.05) but, there was significant difference in control and treatment groups from 3rd month of study. Highest levels were marked in PAM-1 (71.32 ng/ml) and PAM-2 (72.08 ng/ml) groups around 8th month of the experiment. While, highest T4 levels in control (69.06 ng/ml) group was observed around 9th month of the experiment. Treatment groups (PAM-1: 52.22 ng/ml and PAM-2: 51.77 ng/ml) had higher (P < 0.001) overall T4 levels than control (47.61 ng/ml)

group goats.

Nutrient utilization and live weight changes

The present study investigated that PAM-1 and PAM-2 phytoadditive mixtures increased DMI in Black Bengal goats along with improvement in digestibility of nutrients which resulted in higher live weight of supplemented goats. Findings of the present study were in line with the observations of Chaturvedi et al.31 who reported that supplementation of phytogenic mixture containing Tulsi, Turmeric, Amla and Arni (in equal proportions, 0.5% of DM) improved DMI and live weight in supplemented Barbari goats. Further, Usur32 found that supplementation of 3% Garlic powder to Iraqi Black goat resulted in higher DMI and live weight of goats. Redoy et al.33 conducted a randomized growth trial on 32 one year old sheep divided into 4 groups namely, control (no supplementation), PL (plantain herb)- 10 g/kg DM, GL (Garlic leaf)- 10 g/kg DM and PG (plantain herb plus garlic leaves in 1:1)- 10 g/kg DM and observed that DMI, CPI were higher in plantain herb, plantain herb plus garlic leaves in 1:1 than garlic leaves than control groups. Digestibility of DCP, DCF and DEE were similar in all groups but TDN was highest digestible in plantain herb followed by plantain herb plus garlic leaves in 1:1, garlic leaf than control groups. Notwithstanding the findings of this study, a recent study20 investigated that phytogenic mixture containing equal quantities of Zingiber officinali, Allium sativum, Artemisia vulagris and Curcuma longa and commercial tannins @10 g/day/goats showed a non-significant change in feed intake by Shami goats which might be due to incorporation of different phytoadditives. Further, Oderinwale et al.18 showed a non-significant effect of turmeric powder on DMI in goats supplemented (2–5 g/goat/day) in their experiment but, an increase in live weight of supplemented goats. In present study, an additional feeding cost of Rs. 524.0/goat in PAM-1 led to an increase of live weight by 2.30 kg/goat while, Rs. 410.0/goat was incurred in PAM-2 resulted in an increase of live weight by 2.27 kg/goat than control group goats. Considering selling price of Rs. 400.0/kg live weight of goats12, Rs. 920.0/goat was enhanced in value of goats in PAM-1 group and Rs. 908.0/goat in PAM-2 group. Similar to the reporting of this study, Singh et al.12 also found improved live weight of goats supplemented with phytoadditive economically. Earlier studies4,34 reported that supplementation of phytoadditive mixtures improved nutrient digestion through enhanced processes of action of digestive enzymes and reduced ingested nutrient retention time in rumen. The reason behind this may be due to presence of plant secondary metabolites including total phenolics, flavonoids, saponins and alkaloids in Curcuma longa, Allium sativum and Ocimum sanctum favorably modifies rumen function for improved nutrient digestibility in ruminants through anti-microbial properties which leads reduction in methane emissions11,18,21.

Blood indices

The present findings indicated that PAM-1 and PAM-2 phytoadditive mixtures did not affect blood enzymatic, biochemical and hematological attributes of Black Bengal goats. However, blood lipid concentrations were significantly reduced due to phytoadditive mixtures in Black Bengal goats. In corroboration with findings of this study, Usur32 reported a significant reduction in concentrations of blood lipids including cholesterol levels, triglycerides and total lipids in a 3 month trial of supplementing thyme and garlic in Iraqi black goats. Similarly, Thamer et al.35 investigated similar concentrations of WBC, RBC, Hb and PCV % in Awassi lambs supplemented with 2 ml/lamb/day Garlic oil. Hodjatpanah et al.36 showed that addition of Garlic oil (200 mg/sheep/day) or Turmeric powder (20 g/sheep/day) for 28 days didn’t show significant effect on blood glucose and urea levels. Furthermore, Chaturvedi et al.37 also reported that supplementation of Ocimum sanctum, Curcuma longa, Emblica officinalis and Clorodendrium phlomidis in 1:1:1:1 to Jamunapari goats resulted in non-significant effect on blood hematology and biochemical parameters. Similar to present findings, Choubey et al.38 also reported a non-significant effect of phytogenic feed additives that contained tannins and saponins on blood hematology and biochemical parameters. ALT remained unchanged while AST levels were reportedly reduced. Singh et al.39 indicated that lower NEFA levels show lower body fat metabolization and low negative energy balance conditions. Furthermore, Redoy et al.33 found a non-significant effect on blood enzymes such AST and ALP levels and reduced levels of triglycerides and cholesterols in non-descript ewes. The reason behind such reduction in levels of blood lipids might be due to the potential of phytoadditives in decreasing the total lipids, cholesterol and tri glycerides through decreasing the ability of thiol group enzymes such as HMG-CoA and Co-ASH40 in liver while, non-significant effect on other blood parameters showed that supplemented goats were under their normal physiology without adverse effects.

Hormonal profile

This experiment revealed beneficial effect of phytoadditive mixtures (15 g/ kg DM of conc. mixture) containing equal proportions of Curcuma longa and Allium sativum (PAM 1) or Curcuma longa and Ocimum sanctum(PAM 2) on hormonal concentrations in treatment groups. Hormones play a vital role in regulation of physiological and behavioral activities of an animal. Cortisol levels might be helpful in evaluating level of stress whereas tri-idothreonine and thyroxine hormones had roles in body metabolism regulations17. This study showed an increased prolactin levels in treatment group animals which may be due to synergistic effect of plant secondary metabolites in mixture of two herbs. Prolactin hormone is released from anterior pituitary gland that plays essential role in Galactogenesis and Galactopoiesis in goats41. Plant secondary metabolites present in phytoadditives have potential to enhance the release of prolactin through influencing adreno-hypothalamo-hypophyseal-gonadal axis by ceasing the production of dopamine through neurons15,16. Further, Habeeb and EL-Tarabany42 reported that supplementation of Turmeric powder (2 g/goat/day) showed overall increase in T3 and T4 levels with a lower cortisol levels in Zaraibi goats. In contrast with present findings, Oderinwale et al.43 observed a non-significant change in cortisol levels in goats on supplementing them with turmeric powder (2–5 g/goat/day). But, T3 and T4 levels were similar. They remarked that plant secondary metabolites such as total phenolics, alkaloids, saponins and flavonoids had potential to modulate hypothalamic-pituitary-thyroid axis and improved the release of these hormones. In contrast to findings of this study, Hagag et al.44 investigated that 3% garlic powder supplementation in rabbits showed similar level of prolactin hormone. Similar to present findings, Choubey et al.38 investigated a decline in Cortisol levels in goats supplemented with phytogenic feed additive rich in tannins and saponins. Notwithstanding findings of this study, they reported similar T3 and T4 levels. Furthermore, Kumar et al.45 showed that peri-parturient cortisol and T4 levels increase near parturition but, T3 levels were similar. Khan and Ludri46 also reported that concentrations of prolactin and cortisol increases in peri-parturient period whereas, T3 and T4 levels decrease in this period. Also, Singh and Ludri47 found similar results in crossbred goats. Furthermore, Mondal et al.48 have also observed that concentrations of Prolactin and cortisol increases in peri-parturient period whereas, T3 and T4 levels decrease in this period.

Reproductive performance

This study showed favorable effects of phytoadditive mixtures PAM 1 or PAM 2 on reproductive performance of Black Bengal goats through reduced age at first oestrus, first conception, first kidding and post kidding first oestrus days in treatment groups. Varijakshapanicker et al.49 indicated that good reproductive performance in animals showed better animal husbandry practices. Chowdhury et al.50 reviewed that age at first oestrus in Black Bengal goats ranged 131–338 days and age at first kidding was 273–500 days which was similar to the reports of this experiment. More recently, Hossain3 reviewed from several farm level studies that age at first oestrus was 194 to 246 days, gestation period was 143 to 148 days and postpartum first oestrus was observed at 21.43 to 38 days in Black Bengal goats. Chandra et al.13 showed that herbal mixture rich in tannins and saponins improved uterine recovery and delivery process in animals. Similar to findings of this study, Mahour et al.51 observed that supplementation of extracts of Garlic or Tulsi or Turmeric increased conception rates in crossbred cows. Hagag et al.44 remarked that 3% garlic powder supplementation in rabbits improved their reproductive performance through enhanced release of progesterone.

Conclusion

It was concluded from the above study that supplementation of phytoadditive mixtures (15 g/kg DM of conc. mixture) containing equal proportions of Curcuma longa and Allium sativum (PAM 1) or Curcuma longa and Ocimum sanctum (PAM 2) had beneficial effects on reproductive performance, hormonal profile and blood lipid profile through enhanced live weight in female Black Bengal goats.

Acknowledgements

Authors have deep regards toward Indian Council of Agriculture Research, New Delhi for providing necessary funds for this study and Director of National Dairy Research Institute, Karnal and Head of Eastern Regional Station of National Dairy Research Institute, Kalyani for providing all the facilities towards completion of this study. All the persons who were involved directly or in directly in this research are also thanked.

Abbreviations

ALT

Alanine transaminase

AST

Aspartate transaminase and ALP-Alkaline phosphatase

WBC

White blood cells

RBC

Red blood cells

Hb

Hemoglobolin and PCV-Packed cell volume

NEFA

Non esterified fatty acids

T3

Triidothyronine

T4

Thyroxine

Author contributions

AKS, CB and TKD conceptualized the idea. AKS performed and analyzed the data and wrote the manuscript. UK assisted during growth and digestion trial of the experiment. MK and TKD guided and UK helped in laboratory analysis. All the authors considerably contributed to this study and manuscript.

Funding

Authors did not receive any exclusive funding for this study.

Data availability

Data will be made available on reasonable request to corresponding author- Dr. Amit Kumar Singh, amitkumarsingh5496@gmail.com.

Competing interests

The authors declare no competing interests.

Ethical approval

All the experimental procedures were in accordance with relevant guidelines and regulations by ethical committee of the institute (registration: 19/P/LP/09), National Dairy Research Institute, Eastern Regional Station, Kalyani, India.

Footnotes

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Availability Statement

Data will be made available on reasonable request to corresponding author- Dr. Amit Kumar Singh, amitkumarsingh5496@gmail.com.

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