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Research Article

Effect of Maize Silage Inoculated by Bacteria on Performance of Lactating Buffaloes

A.M. Abido, H.S. Soliman , Akila S. Hamza , S.M. Kholif , H.M. Elsayed and A.S. Abd-Elaziz

Two weeks from their parturation, nine lactating buffaloes were used in a 90-d comparative feeding trial to study the effect of uninoculated or inoculated maize silage with Lactic Acid Bacteria (LAB) on their performance. The inoculants were pioneer 1132 which were composed of 100 billion CFU g-1 of crop specific Lactobacillus plantarum and Enterococcus faecium. Animals were similarly grouped into three feeding treatments, according to their weight and milk yield using complete randomized block design. They were introduced to rations containing 60:40 concentrate: roughage ratio as an amount of dry matter equal to 3% of their live body weight. The control group (G1) was fed Concentrate Feed Mixture (CFM) and Rice Straw (RS) as the traditional feeding regimen in summer in Egypt. While, G2 was fed CFM and untreated maize silage and G3 was fed CFM and maize silage inoculated with LAB. Results indicated the digestibility of dry matter, organic matter, crude protein, crude fiber and nitrogen free extract regarding G3 were higher (p>0.05) than those in G1 and G2, while, ether extract digestibility was unaffected by treatments. Milk yield and 4% Fat Corrected Milk (FCM) yield increased significantly with inoculated silage than the other treatments. Milk composition with G3 was higher (p<0.05) for milk total solids, fat and lactose contents than those recorded for G1 and G2. Consequently, G3 produced more yield of milk total solids, fat and protein (p<0.05) than the other groups. Some blood serum parameters were discussed as well. The inoculated maize silage group was better than the other two groups in respect of feed and economic efficiencies.

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A.M. Abido, H.S. Soliman , Akila S. Hamza , S.M. Kholif , H.M. Elsayed and A.S. Abd-Elaziz , 2007. Effect of Maize Silage Inoculated by Bacteria on Performance of Lactating Buffaloes. International Journal of Dairy Science, 2: 330-338.

DOI: 10.3923/ijds.2007.330.338



Many factors related to maize silage preparing. Management can influence silage fermentation characteristics and its quality. Studies have demonstrated some of the chemical changes that occur in the corn plant as it is becoming mature and leads to less fermentable substrates being available for bacterial lactic acid fermentation and thus resulting low silage quality (McDonald et al., 1991). They added Water-Soluble Carbohydrate (WSC) level is the most important substrate related to lactic acid fermentation process. Several studies have demonstrated the effect of bacterial inoculation into corn silage on silage fermentation characteristics (Higginbotham et al., 1998; Cai et al., 1999; Ranjit and Kung, 2000). Earlier criteria for the effective preservation of an ensiled crop included a high degree of lactic acid production and a pH below 4.2 after the fermentation phase (Bolsen et al., 1996; Cleale et al., 1990). Addition of bacterial inoculant containing homofermentative LAB improved the quality and aerobic stability of grass silage (Wrobel and Zastawny, 2004). With maize silage, Jatkauskas and Vrotniakiene (2004) revealed that the inoculant of LAB improved the fermentation quality, increased WSC and lactic acid contents and decreased amounts of acetic acid, butyric acid and ammonia-N. Its application demonstrated an improved intake and milk production with cows, compared to untreated silage. Vrotniakiene and Jatkauskas (2004) concluded that silage treated with the inoculant had a positive effect upon the quality of the fermentation process and positive influence on the bulls growth. Microbial additives improve silage quality, nutrient digestibilities and net energy for lactation (NEL), moreover, they reduce degradation of protein significantly (Pahlow and Honig, 1994; Ilakova et al., 1998).

The objectives of this study were to investigate the effect of inoculated maize silage with LAB on the productive performance of lactating buffaloes.


The present study was carried out in the experimental research station located in Shalakan, Faculty of Agriculture, Ain Shams University, Central laboratory for food and feed, Agriculture Research Center and Dairy Sciences Laboratory, National Research Center.

Experimental Animals
Nine lactating buffaloes at their 3rd lactation season and average weight 527±9 kg were chosen after two weeks of calving form the research station herd. They were divided into three similar groups of three animals each, according to their body weight and milk production, using complete randomized block design. Each group was assigned randomly to one of the three dietary treatments. The experimental animal groups were adapted on the experimental rations two weeks after the calving date and continued up to 105 days of lactation season.

Experimental Rations
The experimental rations were formulated to be 60:40 concentrate: Roughage ratio and offered to animals in order to be dry matter equal to 3% of their body weight. The Concentrate Feed Mixture (CFM) was composed of 30% yellow corn, 35% wheat bran, 25% undecorticated cotton seed meal, 3% rice bran, 3% molasses, 1% urea, 2% limestone and 1% mineral slats.

The control group (G1) was fed CFM and rice straw as the traditional feeding regimen that applied in summer in Egypt. The (G2) was fed (CFM) and untreated maize silage and (G3) was fed (CFM) and maize silage inoculated with lactic acid bacteria. The type of inoculant used was called Pioneer 1132 which were composed of 100 billion CFU g-1 of crop specific Lactobacillus planetarium and Enterococcus faecium. The comparative feeding trail was extended for 90 days preceded by two weeks as adaptation period. The chemical composition of ration ingredients are presented in Table 1.

Animals were individually fed and the daily concentrate feed mixture of each animal was divided into two similar parts. The first was offered before the morning milking (at 6.00 am), while the second one was offered before the evening milking (at 6.00 pm). Rice straw or maize silage was offered to the animals three times a day, at 8.00 am and at 2.00 and 8.00 pm The residues of feed were collected, determined and sampled for chemical analysis. Fresh water was available at all time.

Blocks of Vitamins and minerals mixture structured for dairy cattle were hanged in the front of each animal to be locked freely at all time. Buffaloes were machine milked twice daily at 6.00 am and 6.00 pm and milk yield was recorded for each buffalo.

Table 1: The chemical composition of the CFM, rice straw and uninoculated or inoculated maize silage (% DM basis)
CFM: Concentrate Feed Mixture, RS: Rice Straw, DM: Dry Matter, OM: Organic Matter, CP: Crude Protein, EE: Ether Extract, CF: Crude Fiber, NFE: Nitrogen Free Extract, NDF: Natural Detergent Fiber, ADF: Acid Detergent Fiber, ADL: Acid Detergent Lignin

Feedstuffs and Fecal Analysis
The chemical composition of different feedstuffs and residues were analyzed according to the AOAC (1995) methods for DM, CP, CF, EE and ash, while NFE and OM contents were calculated by difference. Fiber fraction (NDF, ADF and ADL) were determined according to Van Soest and Breston (1979). While, cellulose and hemicellulose were calculated by difference.

Digestibility Trial
Simultaneously three digestibility trail were carried out on all animals of each treatment of feeding trail and repeated each 30 days of the experimental period. Grab sample method was used and silica as internal marker was applied for determining the digestibility. Feces grab samples were collected handily at 8.00 am for three successive days from each animal. Solution of 10% H2SO4 were added to the representative samples then dried in oven at 70°C for 24 h. The dried feces samples from each animal were mixed and stored at -18°C for chemical analysis. The digestibility coefficient was calculated according to the following formula according to Gallup et al. (1945) and Forbes and Garrigus (1948).

Sampling and Analysis of Milk
Milk sample were collected each two weeks during the experimental period. Composite samples were prepared by mixed 1% of each morning and evening milk yield. Milk samples were also, analyzed for fat, total solids (TS), total protein (TP) and ash (Ling, 1963). While, lactose was colorimetric determined according to Barrnett and Abd El-Tawab (1957). Fat Corrected Milk (FCM) was calculated according to Gaines (1928) equation as:

FCM = 0.4 (milk yield kg day-1) + 15 (milk fat yield kg day-1)

Sampling and Analysis of Blood Serum
Blood samples were collected from the jugular vein of each animal at the last day of each period at 4 h post morning feeding. The collected blood samples were centrifuged at 4000 rpm/20 min to separate the serum. The obtained serum was stored at -18°C till it was analyzed. Glucose was determined as described by Trinder (1969) after collecting the sample. Serum total protein was determined as described by Gornall et al. (1949). Albumin was determined as described by Doumas et al. (1971). Gobulin and albumin/globulin ratio were calculated. Serum total cholesterol determination was carried out as described by Allain et al. (1974). Urea was determined by the method of Chaney and Marbach (1962) and Triglycerides was determined by the method of Fossati and Prencipe (1982).

Statistical Analysis
The data were analyzed according to Statistical Analysis System (SAS) User's Guide (1996). Separation among means was carried out by using Duncan (1955) multiple tests used of least square of variance for repeated measures for data of digestibility trails and data of blood serum analysis, while complete randomized block design was used for data of milk yield and composition.


Digestibility Coefficients
The inoculated group G3 was higher (p<0.05) in DM and OM digestibility coefficients then G2 and the later was higher (p<0.05) than G1. Also, significant increases (p<0.05) were noted in CP, CF and NFE digestibility coefficients for the animals fed maize silage (G2 and G3) in comparison with control group (G1). The inoculation had no significant effect on the digestibility of the previous nutrients (G2 and G3) (Table 2). There were significant increases (p<0.05) in Ether Extract (EE) digestibility by G3 when compared to G2 and control group. Several investigators were recorded an improvement in nutrients digestibility due to inclusion of silage ingredient fatting and dairy rations (El-Sayed et al., 1997; Gaafar, 2001; Mostafa et al., 2001). This improvements might be attributed that the reduction occurred in fiber content of hemicellulose during the ensiling process (Dewar et al., 1963; Keady et al., 1996) and a dramatic decrease in ADL content of inoculated silage Table 1. Therefore, it could be affect the digestibility to be better in G3 than G2. Supporting to the findings obtained here, Pahlow and Honig (1994) and Ilakova et al. (1998) found that the inoculation treatment of silage led to an improvement in silage quality and nutrients digestibility. Generally, Davies et al. (1998) concluded that the degree of improvement in nutrient digestibilities of inoculated silage to far extent depend on the availability degree of Water-Soluble Carbohydrates (WSC) in the biomass at silage fermentation phase. Raeth-Knight et al. (2007) found that no differences changes in apparent dry matter, crude protein, neutral detergent fiber, or starch digestibility among treatments when fed dairy cows on Lactobacillus acidophillus and Propionibacteria freudenreichii.

Table 2: Effect of different treatments on nutrients digestibility of lactating buffaloes during 105 day of lactation season
a, b and c Means of treatments within same row with different superscript letter(s) differ (p<0.05)

Blood Serum Parameters
Glucose concentration was significantly higher (p<0.05) for group received corn silage with inoculants (G3) than control group (G1). Also, serum total protein concentration was higher with inoculated silage then other treatments (Table 3). These may be attributed to the higher nutrients digestibility which was recorded for the groups received the maize silage with or without inoculation.

Insignificant differences among treatments were observed in albumin, globulin, urea and total cholesterol. On the other hand, the control group was higher (p<0.05) in triglycerides values than the other groups.

Table 3: Effect of different treatments on blood serum parameters of lactating buffaloes during 105 days of lactation season
a, b and c means of treatments within same row with different superscript differ (p<0.05)

Table 4: Effect of experimental rations on milk yield and composition of lactating buffaloes
a, b and c means of treatments within the same row with different superscript are differ significantly (p<0.05). Ts: Total Solids, TP: Total Protein, SNF: Solids Not Fat

Albumin to globulin ratio ranged between 1.06 and 1.32 and this range are better agreed with that recorded by Khinizy et al. (1997) who fed buffaloe calves on maize silage ration. Moreover, Salem et al. (1989) reported that dietary protein and energy levels are the most effective factors in blood picture. Consistent with this conclusion, Mostafa et al. (2001) revealed that plasma total protein concentration was not affected significantly when grown bulls given isonitrogenous and isoenergetic rations irrespective the differences in ingredients that formulate a certain rations.

Milk Yield and its Composition
Actual milk and 4% FCM yields were significantly higher (p<0.05) for buffaloes fed inoculated silage ration (G3) than those of control (G1) or uninoculated silage (G2). Also, significantly increases in actual milk and insignificantly increases in 4% FCM yields were noted with G2 in relation to G1 Table 4. The higher milk yield with the inoculated silage ration might be attributed to the positive effect of inoculation on the digestibility of organic matter and its nutrients Table 2. These results also probably attributed to the higher of glucose and protein concentration in the blood serum of G3 Table 3. It led to an increase in milk lactose synthesis and consequently milk production being increase. In this respect, Yan et al. (1998) found that inoculation significantly increased the digestibility of organic matter, nitrogen, energy, NDF and ADF in the silage. Therefore, these results were inevitability increased the ruminant production.

Results of milk yield obtained in the present study are in agreement with those obtained by Pahlow and Honig (1994) and Ilakova et al. (1998) who stated that inoculants improved silage quality, nutrients digestibilities and net energy for lactation (NEL). Similar results were obtained by Jatkauskas and Vrotniakiene (2004) who demonstrated that inoculation improved DMI and milk production with cows compared to the uninoculated silage. On the other hand, Kent et al. (1988) and Kung et al. (1992) found no significant effect due to silage inoculants addition. Raeth-Knight et al. (2007) reported no differences effect in average dry matter intake or 4% fat-corrected milk when dairy cows fed Lactobacillus acidophillus and Propionibacteria freudenreichii.

Table 5: Feed efficiency and economic efficiency of the experimental rations
1: Cost of kg FCM produced (LE), 2: FCM/DMI

As an increase of milk yield, daily yields of milk TS, fat and TP were higher (p<0.05) with G3 than those of other groups (Table 4).

Animals fed inoculated maize silage had higher (p<0.05) contents of milk total solids, fat and lactose compared to control group. Also, the TS content was declined (p<0.05), fat did not affected and lactose increased (p<0.05), with G2 comparing with control (G1). No significant differences were reported among treatments with respect to SNF and TP contents. On the other hand, G2 was higher (p<0.05) in ash content than those of both G1 and G3. Results obtained here are in harmony with those found by Holden et al. (1995) and Zaki et al. (2001) who stated that no significant differences in milk constituents due to inclusion different proportions of corn silage in the dairy rations in comparison of the traditional ration that formulated from straw and concentrate mixture. Earlier similar results were obtained by Kent et al. (1988) and Kung et al. (1992). Raeth-Knight et al. (2007) reported that percentage or yield of milk components also did not differed when dairy cows fed Lactobacillus acidophillus and Propionibacteria freudenreichii.

Feed and Economic Efficiencies
Table 5 shows the average amount of daily dry matter feed intake (DMI) in the different groups during the experiment, feed efficiency expressed as kg 4% FCM kg-1 DMI produced and economic efficiency expressed as cost of one kg 4% FCM produced. It appears that the feed efficiency of G3 is better than G2 followed by G1. It seamed that introducing the corn silage to the animals as an excellent roughage source decreased the amount of concentrates consumed by about 20.0% (from 8.9-7.1 kg day-1) and raised the feed efficiency up to about 30.3% (from 0.66-0.86. It led to an increase in milk production up to about 9.85% in G3 and 6.5% in G2 than control. So the economical efficiency was markedly increased with the inoculated silage ration compared with other treatments. Economic efficiency was expressed as cost of feed required to produce one kg fat corrected milk. Assumption price (LE) of one ton of CFM, RS, uninoculated and inoculated maize silage were 1200, 60, 230 and 250, respectively. Brito and Broderick (2006) found that there were linear increases in apparent N efficiency and decreases in N excreted in urine and feces when corn silage replaced alfalfa silage of lactating dairy cows ration. While, Raeth-Knight et al. (2007) reported that feed efficiency averaged 1.52 kg of 4% fat-corrected milk/kg of DM intake and did not differ when dairy cows fed Lactobacillus acidophillus and Propionibacteria freudenreichii.


Using the inoculated whole maize silage as a good roughage source in the ration of lactating buffaloes improved their productive performance and economic efficiency of milk production.


Thanks are the most for both of Dr. Mahmoud Mohammed Ali Khorshid and Mr. Nasr Elbordeny in animal production department, Fac. of Agric., Ain Shams Univ. for their sincere help during collecting data and searching for the references during my acute disease. May ALLAH bless all their doings.

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