Inclusion of different exogenous fibrolytic enzymes to dry jowar fodder and their effect on in vitro total gas production

Aim: Our objective was to estimate in-vitro gas production from dry jowar fodder added with different concentrations of exogenous fibrolytic enzymes (EFEs) like neutral cellulase and fungal xylanase. Materials and Methods: 34 different samples of dry jowar fodder were prepared according to different concentrations of neutral cellulase, fungal xylanase and neutral cellulase + fungal xylanase (1:1). Sample not containing any enzymes was considered as the control group. These 34 samples were subjected to further in vitro gas production analysis. Results: Statistically, significantly higher (P<0.05) potential gas production was recorded for 0.7 % at 6 hr period, 0.7 % at 12 hr period, 0.7 %, 0.8 % at 18 hr period and 0.7 %, 0.8 % at 24 hr period in the samples treated with neutral cellulase. Significantly higher potential gas production was recorded for 0.5 %, 0.8 % at 6 hr period, 0.5 %, 0.6 %, 0.8 % at 12 hr period, 0.8 % at 18 hr period and 0.5 %, 0.6 %, 0.8 % at 24 hr period in the samples treated with fungal xylanase. Significantly higher potential gas production was recorded for 0.6 %, 0.6 %, 0.8 % at 6 hr period, 0.6 %, 0.8 % at 12 hr period, 0.6 %, 0.8 % at 18 hr period and 0.6 %, 0.8 % at 24 hr period in the samples treated with mixture of neutral cellulase + fungal xylanase (1:1). Conclusion: Addition of neutral cellulase and fungal xylanase into the samples of dry jowar fodder increased in vitro total potential gas production. EFEs increase substrate degradation and there by improve the nutritive value of dry jowar fodder.


Introduction
that exogenous enzymes are not effective in the rumen and also that they do not increase the hydrolytic Over the years various forage breeding programs capacity of the rumen.These concerns have been have been used to achieve improved forage quality.disproved by several recent studies which showed that Despite significant genetic improvements, there is supplementation of EFEs to ruminant feeds increase excessive loss of nutrients by ruminants due to their nutritive value and consequently improve the limitations in forage degradability.To improve the animal's productivity [6,7].forage degradability various physical, chemical and Addition of EFEs increases microbial growth and biological treatments are routinely used.The use of increases the production of microbial proteins [8].exogenous fibrolytic enzymes (EFEs) to improve Addition of fibrolytic enzymes to feedstuffs increased forage degradability holds promise as a means of dry matter (DM), crude protein (CP), and soluble improving the productive efficiency of ruminants.
fractions of diets, -an affect that provides energy and Action of EFEs on feedstuffs can release more also leads to rapid microbial growth [9].Increased nutrients, because EFE can break down specific number of ruminal bacteria could then lead to chemical bonds in the feedstuffs that are not usually increased microbial colonization of the feed particles degraded by endogenous enzymes [1].EFEs increase which can result in increased total gas production.digestion of dry matter and fiber as measured in situ Jowar fodder is commonly used by farmers of [2], in vitro [3,4]  In-vitro Gujarat.The activities of the enzymes were 2,00,000 medium was prepared by mixing 300 ml distilled units/gm and 3,000 units/gm for fungal xylanase and water, 200 ml rumen buffer solution and 100 ml macro neutral cellulase, respectively.minerals solution, 0.4 ml Resazurine solution, 0.04 ml micro minerals solution and 22 ml reducing solution Sample preparation: Sample of dry jowar fodder was (prepared fresh and added just prior to incubation).The collected from cattle breeding farm, Junagadh.Fully o medium was pre-warmed to 39 C and bubbled with matured sample of jowar fodder was used for this CO till the blue colour of the medium disappeared.research.Jowar fodder was harvested by labor after 2.5 2 After the medium became colorless, 200 ml of strained month of maturity period.No machines were used for rumen liquor (SRL) was added.Then 40 ml of harvesting the Jowar fodder.Sample was dried in hot o incubation medium was injected into each syringe air oven at 70 C for overnight.Sample was ground to reduce particle size (0.5mm) and stored in polythene using pipette.Level of piston was recorded and the o o bag at room temperature (37 C)nalysis.Eleven different syringes were placed in an incubator (39 ± 0.5 C).Gas samples of dry jowar fodder were prepared according produced (ml/ 500 mg substrate) during fermentation to different levels of neutral cellulase i.e. 0.01, 0.1, 0.2, was measured after 24 hrs.0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 and 1%.Eleven different Statistical analysis: Experimental data were analyzed samples of dry jowar fodder were prepared according as per CRD [14].The treatment means were compared to different levels of fungal xylanase i.e. 0.01, 0.1, 0.2, by Duncan's new multiple range test [15].P-values 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 and 1%.Eleven different less than 0.05 are considered to be statistically samples of dry jowar fodder were prepared according significant.to different levels of neutral cellulase + fungal xylanase (1:1) i.e. 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9

and 1%. Sample of dry jowar fodder not containing any
The gas production technique has been widely of the enzymes was considered as control.These 34 used for evaluation of nutritive value particularly for samples of dry jowar fodder were subjected to in vitro various types of tropical plants [16,17] and for different analysis.
classes of feeds [18].In vitro gas production technique Estimation of total gas production: Modified method helps to quantify nutrient utilization in a better way by was used for estimation of total gas production [10,11].
its accuracy in describing digestibility in animals [19].

In this method increasing the amount of sample from
This technique can be used to predict animal's 200 to 500 mg and increasing the amount of buffer twoperformance at a much lower cost.In this study fold as a result the incubation volume increase from 30 significant effects of EFE treatment on fractional gas ml in the method [12] to 40 ml in the modified method.production at four different periods i.e. 6 hr, 12 hr, 18 hr 500 mg sample quantity was chosen to decrease and 24 hr periods of incubation was recorded.Effect of analytical error but at the same time our goal was to different levels of neutral cellulase enzyme on in vitro avoid production of more than 90 ml gas [13].
gas production from dry jowar fodder is shown in Table-1.Experimental substrate (500mg) was weighed on Our results show that gas production parameters from a plastic boat with removable stem and placed into the all treatment groups were significantly (P<0.05)bottom of the glass syringe without sticking to the sides comparable with those of the control group, except for neutral cellulase enzyme level at 0.01 % in dry jowar of the syringes.The piston was lubricated with study.This may be due to differences in the enzyme potential gas production was recorded for 0.7 % at 6 hr activity and samples used for in vitro analysis.period, 0.7 % at 12 hr period, 0.7 %, 0.8 % at 18 hr Effect of different levels of neutral cellulase + period and 0.7 %, 0.8 % at 24 hr period.Effect of fungal xylanase (1:1) enzyme on in vitro gas different levels of fungal xylanase enzyme on in vitro production from dry jowar fodder, at four different gas production from dry jowar fodder, at four different periods is shown in Table -3.Results show that gas periods is shown in Table-2.Results show that gas production parameters from all treatment groups were production parameters from all treatment groups were significantly comparable with those of control group.significantly comparable with those of control group.
Significantly higher potential gas production was Data indicated that total potential gas production from recorded for 0.6 %, 0.6 %, 0.8 % at 6 hr period, 0.6 %, samples treated with fungal xylanase is higher than the 0.8 % at 12 hr period, 0.6 %, 0.8 % at 18 hr period and samples treated with neutral cellulase.Results indicated 0.6 %, 0.8 % at 24 hr period.Results from Table 1 to 3 that up to a certain level, increasing concentrations of indicated that highest amount of total potential gas EFEs also increased in vitro total gas production production is recorded for samples treated with which may be due to increased enzymatic activity at mixture of neutral cellulase + fungal xylanase (1:1) and higher concentrations of EFEs.
lowest amount of total potential gas production is Six enzyme products were used to examine the observed in samples treated with neutral cellulase.Our relationship between enzyme activities and in vitro gas results indicate that effect of different levels of neutral production using grass and corn silage [20].
cellulase + fungal xylanase (1:1) enzyme on in vitro gas Preparations relatively high in cellulase activity production from dry jowar fodder is higher than the increased the rate of gas production from corn silage effects of neutral cellulase and fungal xylanase compared with the control (no added enzyme).In individually.Higher gas production data from the contrast, products with relatively high xylanase samples containing mixture of neutral cellulase + activity did not increase gas production when fungal xylanase (1:1) indicated that a combination of glucanase activity was low [20].Those results were both the enzymes is beneficial rather than their similar to the effects of neutral cellulase, but individual use to improve total gas production.contradictory to the effects of fungal xylanase in this The addition of neutral cellulase, fungal xylanase and in sacco[5].Previously EFEs are Saurashtra region in Gujarat.Hence this research was only used to improve the nutritive value of feeds for performed to evaluate the effect of different EFEs on non ruminants particularly in broiler diets.In ruminant the in vitro digestibility of dry jowar fodder.feeds EFEs were not used because of the perception

Table - 2
. Effect of fungal xylanase on in vitro gas production (ml) from dry jowar Means in a column with different superscripts indicate significant (P<0.05)difference. *

Table - 3
. Effect of mixture of neutral cellulase + fungal xylanase (1:1) on in vitro gas production (ml) from dry jowar * Means in a column with different superscripts indicate significant (P<0.05)difference.