A Study on the Effect of Ration Balancing for Improving Milk Production and Reducing Methane Emission in Lactating Buffaloes under Field Conditions

Indian Journal of Dairy Science, 65(3): 250-255 (2012)

A Study on the Effect of Ration Balancing for Improving Milk Production and Reducing Methane Emission in Lactating Buffaloes under Field Conditions

                     M R Garg*, A Kannan, B T Phondba, S K Shelke and P L Sherasia

Animal Nutrition Group,

National Dairy Development Board,

Anand- 388 001, Gujarat, India


A field trial on twenty six lactating buffaloes was conducted to study the effect of ration balancing on milk production, microbial protein synthesis and methane emission. Baseline methane emissions of buffaloes were estimated by using SF6 tracer technique, thereafter the ration was balanced as per their nutrient requirements. After 30 days of feeding balanced ration, methane emission by the animals was estimated again. Microbial protein synthesis was calculated by estimation of purine derivatives in urine. Analysis of the feeding practices revealed that though the dietary intake of CP was adequate, TDN intake was lower in buffaloes (14.48%) than their requirement. The calcium and phosphorus were also deficient by 43.58% and 76.66% in buffaloes. On feeding balanced ration, average increase in milk yield (kg/day), fat (%), 6% FCM yield (kg/day) and microbial nitrogen supply (g/day) in lactating buffaloes were 0.46, 0.29, 0.71 and 29.18, respectively, which were higher (P<0.01) than before implementation of the programme. Average methane emission, in terms of g/day was reduced (P<0.01) by 13.30% after feeding balanced ration. Similarly, methane emission in terms of g/kg DM intake, g/kg OM intake and g/kg milk yield were also reduced (P<0.01). The gross energy lost as methane was reduced (@ 18.13%; P<0.01) in buffaloes after balancing the ration. Thus, the results of the present study indicated that, ration balancing has the potential for improving milk production, milk fat and microbial nitrogen supply along with reducing methane emission in lactating buffaloes under field conditions.


Keywords: Ration Balancing, Milk Production, Methane Emission, Buffaloes, Field Study




India possesses the world’s largest livestock population of 485 million (13% of global livestock) and has 57% of the world’s buffalo population. Currently, it is the largest milk producing country in the world with 112.5 million tones of milk production for the year 2009-10 (MOF, 2010). Buffalo is the main milk producing animal which contributes 58% of the total milk production in India with an average lactation yield of 1300 kg (GOI, 2007). This low yield is mainly due to feeding of poor quality feed resources (crop residues and agro industrial by-products) to buffaloes domesticated in rural households.


Enteric fermentation emitted 10.09 million tonnes of methane and is responsible for 73.3% of total methane emission from agriculture sector in India (INCCA, 2010). Buffalo is the single largest emitter of methane due to its higher methane emission coefficient i.e. 50 kg/animal/year (NATCOM, 2004) and constitutes 42% of the total methane emission from livestock sector for the year 2003 (Chhabra et al. 2009). Microbial fermentation of feeds and fodder in rumen results in production of VFA, microbial protein, H2 and CO2. The major producers of H2 are the organisms which produce acetate in the fermentation pathway. Acetate and butyrate formation is responsible for enteric methane production, while propionate formation serves as competitive pathway for H2 utilization.  Under Indian feeding situations, where poor quality roughages forms major constituent of animal diet, production of acetate is more which contributes to higher methane production and reduced microbial protein synthesis. Also in field conditions, animals are not fed as per their nutrient requirement which leads to poor productivity of animals. The substantial increase in the productivity of ruminants on low digestible forage based diets can be achieved through balanced nutrient approach that considers the efficiency of the rumen ecosystem and the availability of the dietary nutrients post-ruminally. Keeping these points in view, the present experiment was designed to study the effect of ration balancing on milk production, microbial protein synthesis and methane emission in lactating buffaloes under field conditions.



Experimental Design

Twenty six lactating buffaloes belonging to twenty two farmers were chosen for the study from seven villages of Nanded district in Maharashtra state (India). The selected buffaloes were in their third lactation, with an average milk production of 6.10 kg per day and 6.48% milk fat. The feed was offered by the farmers twice a day i.e. morning and evening. The feed intake of individual animal was measured and representative sample was taken for proximate principles. Thereafter, the ration of all animals was balanced for total digestible nutrients (TDN), crude protein (CP), calcium and phosphorus using the ration balancing software developed by National Dairy Development Board (NDDB), which is based on Kearl (1982) standards for buffaloes. The balanced diet was fed to all the animals for 30 days. Urine (100 ml) samples were collected from individual buffaloes before and after balancing the ration and preserved with sufficient quantity of 10% H2SO4 to maintain pH below 3. The urine samples were diluted in such a way that the concentration in the final sample would fall within the range of standards used in the assays for estimation of purine derivatives (IAEA, 1997). The body weights of the animals were calculated using Shaeffers’s formula: Body weight (kg) = ([(heart girth in inches)2 x length of the body in inches]/300) x 0.4536.

Methane Measurement

Dry matter (DM) intake, milk yield and milk fat were recorded daily during methane gas sampling period. Methane measurement was done by sulfur hexafluoride (SF6) tracer technique (Johnson et al. 1994). A small permeation tube containing SF6 was inserted in the rumen of each of the experimental animal. The breath samples of all experimental animals were collected daily for four consecutive days in canisters and brought to the laboratory for methane and SF6 analysis at the start of study. After one month of experimental feeding, the methane emission was measured similarly. Methane and SF6 concentrations were determined by gas chromatography. Methane emission rate was calculated as the product of the permeation tube emission rate and the ratio of CH4 to SF6 concentration in the sample. Samples were analyzed in duplicate. The gas chromatograph was fitted with a molecular sieve 5A column for SF6 and a Porapack N for methane. The column temperature was maintained at 500C and nitrogen was used as a carrier gas, with flow rate of 30 ml/min. Prepared standards were used to standardize the gas chromatograph for SF6 (39.2 ppt and 101.7 ppt, Scott-Marrin Inc., Riverside, CA, USA) and methane (10.4 ppm and 101.9 ppm, Scott-Marrin Inc., Riverside, CA, USA).

Methane emission rate was calculated as under:

Q CH4 = Q SF6 x (CH4) / (SF6)

Where, Q CH4 = Methane emission rate (g/min)

             Q SF6 = Known release rate of SF6 from permeation tube (g/min)

             CH4 = Methane concentration of collected sample in canister (µg/m3)

             SF6 = SF6 concentration of collected sample in canister (µg/m3)


Laboratory Analysis

Feeds and fodder samples were analyzed for proximate composition by AOAC (2005) methods. The milk samples were analyzed for milk fat (IS: 1224, 1977) and 6% FCM yield was calculated. The urine samples were assayed for allantoin, uric acid and creatinine (Young and Conway, 1942; Hawk et al. 1976). Purines absorbed and microbial nitrogen supply was calculated from the daily urinary purine derivatives (PD) excreted (IAEA, 1997). Gross energy of feed and fodder samples were calculated as per the prediction equation of Guenther (1979). Energy content of methane was taken as 13.34 kcal/g (Brouwer, 1965). The data were statistically analyzed using paired student’s t-test (Snedecor and Cochran, 1994).


Feeding Practices and Ration Balancing

From feeding practices, it was observed that cottonseed cake was the most commonly used protein source for dairy animals by the farmers in Nanded district of Maharashtra. Farmers used mixed local grasses, maize fodder, jowar fodder, guinea grass, hybrid napier and sugarcane tops as green fodder. Jowar and soybean straw were used as dry roughage. Feeding of mineral mixture to the animals was not practiced. Body weight and feed intake of the experimental animals before and after ration balancing is presented in Table 1. Analysis of feeding practices revealed that though the dietary intake of CP was adequate, TDN intake was lower (14.48%) than their requirements. The calcium and phosphorus were also deficient by 43.58 and 76.66%, respectively. Tiwary et al. (2007) and Yadav et al. (2002) reported deficiency of CP, TDN, Ca and P in lactating buffaloes under Indian field conditions. Kannan and Garg (2009) also reported deficiency of CP, Ca and P under field conditions in the states of Gujarat and Uttar Pradesh.

After the collection of samples for baseline methane emission, the ration was balanced as per the requirement of individual animal for TDN, CP, Ca and P.  While balancing the ration, major emphasis was given on energy sources to counteract the TDN deficiency. To fulfill the Ca and P requirements, mineral mixture was incorporated in the ration. After balancing the ration, the intake of DM (kg/day, percentage of body weight and per unit metabolic body size) was higher (P<0.01). The TDN intake (g/day) was also improved (5455.85 vs 6401.77; P<0.05). The concentrate to roughage ratio was increased due to incorporation of energy sources in the ration. Balancing the ration also increased (P<0.01) the body weight and metabolic body size of buffaloes. The intakes of dry matter (kg/day, percentage body weight and per unit metabolic body size) and TDN (g/day), body weight and ratio of concentrate to roughage was similar to those reported for buffaloes by Paul et al. (2003). Srinivas and Singh (2010) also reported higher intake of energy after increasing the proportion of concentrate in the ration of ruminants.   

Effect of Ration Balancing on Milk Yield, Milk Fat and FCM Yield

Ration balancing improved (P<0.01) milk yield (kg/day), milk fat (%) and 6% FCM by 0.46, 0.29 and 0.71, respectively (Table 1). The improvement in milk yield, milk fat and 6% FCM may be due to balancing of nutrients, which might have alleviated the energy deficiency in terms of TDN and also due to supply of mineral mixture which might have alleviated the deficiency of calcium and phosphorus. Findings are similar to those of Haldar and Rai (2003) who reported improvement in milk yield due to supplementation of energy and mineral mixture in lactating ruminants. Energy is one of the most important limiting factors towards milk production and its supplementation in the diets of lactating ruminants increased milk yield significantly. Another important aspect in the physiology of lactation is the severe drainage of minerals through milk. Supplementation of minerals in the diet of lactating ruminants has been reported to enhance milk production along with an improvement in milk composition (Kannan et al. 2010).

Table 1: Body Weight, Plane of Nutrition and Milk Yield of Experimental Buffaloes



After RBP

Body weight (kg)

451.15c ± 12.31

457.04d ± 12.33

DM intake (kg/d)

10.27c ± 0.23

11.63d ± 0.28

Concentrate: Roughage ratio



DM intake (kg/100 kg B. wt.)

2.27c ± 0.06

2.54d ± 0.05

DM intake (g/kg W0.75)

104.91c ± 0.002

117.65d ± 0.002

CP intake (g/d)

1239.85 ± 90.69

1256.00 ± 40.36

TDN intake (g/d)

5455.85a ± 364.07

6401.77b ± 228.12

N intake (g/kg digestible OM intake)

20.89 ± 1.70

22.85 ± 0.89

Milk yield (kg/d)

6.10c ± 0.35

6.56d ± 0.37

Milk fat (%)

6.48c ± 0.16

6.77d ± 0.13

6% FCM yield (kg/d)

6.44c ± 0.38

7.15d ± 0.43

                a, b Values with different superscript in a row differ significantly (P<0.05).

c, d Values with different superscript in a row differ significantly (P<0.01).


Purine Derivatives Excretion and Microbial Nitrogen Supply

The mean concentration of purine derivatives (PDC) are presented in Table 2.  Balancing the ration of lactating buffaloes resulted in improvement (P<0.01) in PDC index, PD excreted and absorbed purines thus, improved by 39.35% microbial nitrogen supply to buffaloes. Microbial protein synthesis in rumen depends upon supply of ammonia, energy and carbon skeleton for amino acid synthesis (Tomar et al. 2010). Most of the carbon skeletons are produced as a result of degradation of carbohydrates into VFAs. In the present study, the diet of buffaloes was adequate in CP but deficient in TDN.  In such condition, the inadequate supply of energy might be responsible for poor availability of ATP and carbon skeleton for microbial cell production thereby reducing microbial protein synthesis. After balancing the ration of buffaloes, greater availability of energy might have resulted in increased microbial protein synthesis thereby improving the performance of buffaloes. The present findings are similar to Srinivas and Singh (2010) and Ramgaonkar et al. (2008) who reported an increased excretion of PD and microbial nitrogen supply after supplementing high plane of nutrition in ruminants.

Table 2: Effect of Ration Balancing on Rumen Microbial Protein Synthesis



After RBP

Average metabolic body weight (kg w0.75)

97.89c ± 1.95

98.85d ± 1.94

Uric acid (mmol/l)

0.24 ± 0.001

0.24 ± 0.001

Allantoin (mmol/l)

2.81 ± 0.23

3.02 ± 0.25

Purine derivatives concentration (mmol/l)

3.05 ± 0.23

3.26 ± 0.26

Creatinine concentration (mmol/l)

2.90 ± 0.23

2.33 ± 0.16

PDC index

103.13c ± 5.03

138.10d ± 12.38

Total PD excreted (mmol/d)

101.07c ± 4.93

135.33d ± 12.13

Absorbed purine (mmol/d)

101.98c ± 5.79

142.12d ± 14.04

Intestinal flow of microbial nitrogen (g/d)

74.14c ± 4.21

103.32d ± 10.21

c, d Values with different superscript in a row differ significantly (P<0.01).


Effect of Ration Balancing on Methane Emission

Baseline methane emission from buffaloes was 154.47 g/animal/day (Table 3). After feeding balanced ration the methane emission was reduced (P<0.01) to 133.92 g/animal/day. Thus, there was 13.30% reduction in methane emission from lactating buffaloes after balancing their ration. Methane emission (g/kg DM intake) at baseline and after balancing the ration was 15.04 and 11.51, respectively. Similar values are also reported by Mohini and Singh (2001). Methane emission in terms of g/kg DM intake, g/kg OM intake and g/kg milk yield were reduced (P<0.01) than the baseline emissions. The gross energy intake (Mcal/d) was increased (P<0.01) after balancing the ration of buffaloes. The energy lost as methane was 2.06 and 1.78 Mcal/d at baseline and after balancing the ration, respectively and was reduced (P<0.01). Due to imbalanced ration, there was loss of 5.02% gross energy from buffaloes and after feeding balanced ration, it was reduced (P<0.01) to 4.11%.

The baseline methane emission observed in present study was similar to Condor et al. (2008). Reduction in methane emission after balancing the ration was similar to Mohini and Singh (2010) who observed that changing plane of nutrition through concentrate mixture or urea molasses mineral block, improved digestibility of nutrients as well as decreased methane production in lactating cows. The reduction in loss of gross energy through methane in present study is consistent with earlier reports (Ominski et al. 2004). In the present study, due to balancing of nutrients, the rumen fermentation pattern of buffaloes might have changed towards more microbial cell production, lower volatile fatty acids (acetate and butyrate) production and lower methane emission. In ruminants, feed nutrients provide both the substrate for microbial cell synthesis and also the potential energy as ATP generated through conversion of feed biomass to VFA. On imbalanced diet, the yield of microbes relative to VFA produced is variable, i.e. ATP is used with variable efficiency which is closely related to methane emission. High proportional feed conversion into microbial protein synthesis, that is a high efficiency of microbial production and high proportion of propionate, reduce digestive carbon losses (Blummel et al. 2010).


Table 3: Effect of Ration Balancing on Methane Emission



After RBP

Methane emission (g/animal/d)

154.47c ± 5.46

133.92d ± 5.37

DM intake (kg/d)

10.27c ± 0.23

11.63d ± 0.28

Methane emission (g/kg DM intake)

15.04c ± 0.63

11.51d ± 0.51

OM intake (kg/d)

9.68 ± 0.20

10.02 ± 0.41

Methane emission (g/kg OM intake)

15.95c ± 0.74

13.36d ± 0.64

Methane emission (g/kg milk yield)

25.32c ± 1.59

20.41d ± 1.35

Gross energy intake (Mcal/d)

41.01c ± 1.33

43.24d ± 1.34

Energy loss as methane (Mcal/d)

2.06c ± 0.07

1.78d ± 0.07

Energy loss as methane (% of gross energy)

5.02c ± 0.24

4.11d ± 0.21


c, d Values with different superscript in a row differ significantly (P<0.01).



From the present study, it can be concluded that ration balancing programme has the potential to improve milk production and reduce methane emission in lactating buffaloes. More research trials may be needed to document the methane emission from lactating animals under diverse feeding practices followed in different agro-climatic regions of India. There is scope for improving feeding pattern within limited available feed resources through implementation of ration balancing programme for improving productivity and reducing methane emission in environment friendly manner.



Financial assistance and facilities provided by the management of National Dairy Development Board, Anand, for undertaking this study, are gratefully acknowledged.





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