A growth trail was conducted with 24 Hariana cattle heifers of 12-16 months old, weighing about 150 kg to study the effect of feeding formaldehyde treated mustard oil cake based diet on feed intake, growth performance, nutrient utilization, certain mineral absorption and serum biochemical parameters. Animals were randomly divided into four groups of 6 animals each and were individually fed for a period of 120 days. The animals in group I was fed with diet containing 80% crude protein of NRC, group II were fed with diet containing 100% crude protein of NRC, group III were fed with formaldehyde treated mustard cake diet containing 80% crude protein of NRC and group IV were fed with diet containing formaldehyde treated mineral fortified mustard cake containing 80% crude protein of NRC. Average daily body weight gain (g/d) of animals in group IV was significantly (p<0.05) higher in comparison to group I, but statistically similar with group II. Absorption of minerals (Ca, Mg, Cu, Fe, Zn, Mn and Se) were statistically similar except P, which was significant (P<0.01) higher in group II than that of other groups. Higher concentration of serum Zn in group II, while the serum Ca level increased significantly for all the groups as the trial progressed. Incorporation of formaldehyde treated mustard oil cake in the concentrate mixture led to increase in growth rate by 23.5% due to better balance of nutrients. Thus, it may be concluded that formaldehyde treatment of mustard cake is beneficial in growing heifers without any stress as assessed by serum biochemical constituents, enzymes activities and mineral profile.
In tropical countries like India, oilseed cakes constitute the major protein sources in ration of dairy animals (Sahoo et al., 2006). India is the second largest producer of rape seed mustard in the world and its cake is one of the by-products available at cheaper price throughout the year (Anil Kumar et al., 2002; Sahoo et al., 2006). It is rich in some of the essential amino acids such as methionine and lysine but its nutritive value is reduced due to high rumen degradability (Sahoo and Walli, 2008). This rapid and high ruminal degradation of rapeseed mustard cake protein signifies the need of research for protection of the protein from degradation and the wastage. Bypass protein technology like formaldehyde treatment protect the proteolysis and allow these valuable proteins to bypass rumen and get absorb in intestine (Kanjanapruthipong et al., 2002; Wulf and Sudekum, 2005; Walli, 2005; Tiwari et al., 2010; Parnerkar et al., 2010; Yadav and Chaudhary, 2010). Formaldehyde treatment have been used by different workers to protect the protein from ruminal degradation in India and abroad but the information on feeding of formaldehyde treated mustard oil cake to indigenous growing cattle heifers is lacking. The present study was conducted to study the effects of feeding formaldehyde treated rape seed oil cake on growth performance, nutrient utilization and mineral bioavailability and evaluate this as a protein source in growing hariana heifers.
Materials and Methods
Formaldehyde Treatment of Rape Seed Mustard Oil Cake
Rape seed oil cake was taken from a local market of Mathura, Uttar Pradesh and ground to pass 1.0 mm sieve size and subjected to crude protein (CP) estimation by Kjeldahl’s technique. Formaldehyde treatment for protection of mustard cake was conducted as per the procedure given by Sahoo et al. (2006). The treatment was conducted in the feed manufacturing unit of the dairy farm of DUVASU and the 40% formaldehyde was sprinkled over the ground cake (mustard cake without mineral mixture and with mineral mixture) in horizontal mixer to have a final concentration of 1.5% per 100 g of crude protein of mustard cake. The treated cake samples were mixed thoroughly and stored in tightly sealed plastic bags and then air tight plastic containers were stores at room temperature (25°C) for 7 days to make protected protein. After that treated mustard cake was stored in air tight plastic boxes until use. Treated mustard cake was added to concentrate in appropriate amount to make concentrate ration.
Housing, Management and Selection of Experimental Animals
The experiment was conducted in Instructional Livestock Farm Complex (ILFC, 27°27’52″N and 77°42’26″E) at Uttar Pradesh Pandit Deen Dayal Upadhayay Pashu Chikitsa Vigyan Vishwavidyalay Evum Go-Anusandhan Sansthan (DUVASU), Mathura, India. Permission of taking animals for this experiment was duly taken from Institutional Animal Ethics committee (IAEC) constituted as per the Article No. 13 of the CPCSEA rules laid down by Government of India. Twenty four healthy growing Hariana heifers of 12 to 16 months old weighing approximately 150 kg were randomly assigned into four groups comprising each of six animals. Before starting the experiment, all the animals were dewormed against internal parasites by recommended doses. Animal were stall fed individually ad-libitum and weighed quantities of rations were offered once a day at 09:00 am. The animals were fed four different diets satisfying the requirement of NRC, 2001. Group I was fed with diet containing 80% crude protein of NRC, group II with diet containing 100% crude protein of NRC, group III with formaldehyde treated mustard cake diet containing 80% crude protein of NRC and group IV with diet containing formaldehyde treated mineral fortified mustard cake containing 80% crude protein of NRC. Concentrate mixture consisting of 55% barley, 10% wheat bran, 33% rape seed mustard oil cake, 2% mineral mixture and 1% salt were prepared.
120 days of growth trial was conducted. A digestion trial of seven days duration for individual animals was conducted after end of growth trial. Feed samples, residues and faecal output were collected at 24hrs intervals and sampled for DM estimation to assess feed and DM intake in animals during the experimental feeding. The body weight was recorded for two consecutive days fortnightly before offering feed and water throughout the trial.
Dried samples of feed offered, residues and faeces were collected during the trial and analyzed for dry matter (DM), organic matter (OM), crude protein (CP), ether extract (EE) as per the standard procedure (AOAC, 2000), while the fibre fractions (NDF and ADF) were analysed as per Van Soest et al. (1991). The concentration of major and trace mineral in feeds and fodders wereestimated with the help of Atomic Absorption Spectrometer (Perkin Elmer, USA), by using flame and graphite mode. Ground samples of feed, fodder, residue and faeces sample were digested with the help of triple acid solution (nitric acid, sulphuric acid, perchloric acid in ratio of 3:1:1) and volume was made to 25 ml before analysing by AAS. However phosphorus content of samples was estimated as per method described in AOAC (1995).
Biochemical and Serum Analysis
Blood samples were collected aseptically from jugular vein of each growing Hariana heifer on 0, 30th, 60th, 90th and 120th day of the experimental feeding trial at 8.00 AM prior to morning feeding and watering and serum were separated and stored at -20ºC till use. The serum samples were analyzed in chemistry autoanalyzer (Mindray) using ready-to-analyze kits provided for serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) activities and serum concentrations of calcium, phosphorus, magnesium, total protein, albumin, creatinine, urea and cholesterol. Serum globulin content was calculated by subtracting albumin content from the total protein content. Trace minerals like copper, iron, zinc, manganese and selenium were estimated by with the help of Atomic Absorption Spectrometer (Perkin Elmer, USA), by using flame and graphite mode.
The data obtained was processed to analysis of variance as per standard method of Snedecor and Cochran (1994), and means and homogenous subsets were separated by using Duncans multiple range test described by Duncan (1955).
The chemical composition of feed ingredient during feeding trials and ingredients (%) and dietary composition of dietary treatments fed during feeding trial to experimental heifers are presented in Table 1 and 2, respectively.
Table 1: Chemical composition (%DM basis) of feed ingredient during feeding trial
|Attributes||Concentrate Mixture||Berseem Fodder||Wheat Straw|
|Acid Insoluble Ash||0.41||0.10||0.57|
|Nitrogen Free Extract||52.49||36.31||26.76|
|Nutrient Detergent Fibre||38.6||56.09||81.88|
|Acid Detergent Fibre||13.61||42.13||53.9|
|Acid Detergent Lignin||1.9||8.66||8.19|
Table 2: Ingredients (%) and dietary composition of dietary treatments fed during feeding trial to experimental heifers
|Composition (%DM basis)|
|Nitrogen Free Extract||40.39||43.29||40.39||40.39|
|Neutral Detergent Fibre||55.54||51.62||55.54||55.54|
|Acid Detergent Fibre||33.66||28.79||33.66||33.66|
|Acid Detergent Lignin||5.83||4.84||5.83||5.83|
I: Negative control (diet containing 80% crude protein of NRC); II: Positive control (diet containing 100% crude protein of NRC); II: Formaldehyde treated mustard cake (diet containing 80% crude protein of NRC); IV: Formaldehyde treated mineral fortified mustard cake (diet containing 80% crude protein of NRC)
The mean nutrient intake and digestibility of nutrients are shown in Table 3 which showed DM, CP and total digestible nutrients (TDN) intake (Table 3) on metabolic body size basis were similar among the different groups with similar digestibility except OM, EE, nitrogen free extract (NFE) and neutral detergent fibre (NDF). The average total DMI (kg/d), DM intake (kg/d) on percent body weight basis, metabolic body weight fortnightly (kgW0.75), feed conversion ratio (kg DMI/ kg weight gain) during the growth trial were similar in all the groups (Table 4). The assimilation of minerals viz. Ca, Mg, Cu, Fe, Zn, Mn and Se absorption among the groups were statistically similar, while absorption of P was significant higher (P<0.01) in group II than that of other groups (Table 5). The values for serum biochemical parameters are given in Table 6. There was no significant difference in serum creatinine (mg/dl), cholesterol (mg/dl), blood urea (mg/ dl), serum total protein, albumin, globulin, SGOT and SGPT activity among different groups of heifers.
Table 3: Nutrient intake and digestibility coefficient of nutrients during digestion trial
|Attributes||Treatment Group||Overall Mean||SEM||P-value|
|Plane of Nutrition|
|g/kg W 0.75||95.13||93.24||95.23||105.61||97.30||2.05||0.12|
|g/kg W 0.75||296.48||315.11||320.12||341.04||318.19||6.36||0.09|
|g/kg W 0.75||1.59||1.44||1.52||1.72||1.57||0.05||0.21|
|Digestibility Coefficients (%)|
|Crude Protein (CP)||75.69||78.99||78.95||79.36||78.24||0.67||0.18|
|Ether Extract (EE)||55.78a||71.05c||73.33c||63.63b||65.95||1.99||0.002|
|Crude Fibre (CF)||48.14||49.61||50.42||53.78||50.42||1.17||0.39|
|Nitrogen Free Extract (NFE)||84.33b||73.03a||82.03b||82.95b||80.08||1.31||0.001|
|Nutrient Detergent Fibre (NDF)||59.59b||53.89a||59.48b||62.77c||58.93||0.77||0.001|
|Acid Detergent Fibre (ADF)||44.98||37.4||39.92||40.28||40.65||1.03||0.05|
Mean bearing different superscript in a row differed significantly (P<0.05)
Table 4: Absorption (%) of minerals fed to experimental Hariana heifers during digestibility trial
|Attributes||Treatment group||Overall mean||SEM||P- value|
Mean bearing different superscript in a row differed significantly (P<0.05)
Table 5: Growth rate and feed conversion ratio in Hariana heifers during growth trial
|Fortnights||Treatment Group||Overall Mean||SEM||P-value|
|Initial body weight (kg)||154.80||155.90||154.80||155.13||155.06||5.63||1.00|
|Final body weight (kg)||205.68||212.55||217.49||213.72||212.36||5.70||0.91|
|Average live weight gain(g/day)||424a||472ab||525b||488ab||477||15.0||0.04|
|Metabolic body weight fortnightly (kgW0.75)||48.98||49.75||49.87||49.73||49.58||1.16||0.99|
|Average DM intake (Kg/day)||4.87||5.01||4.82||5.01||4.90||0.15||0.95|
|DM Intake on percent
body wt. basis (kg)
|Feed conversion ratio
(kg DMI/ kg weight gain)
Mean bearing different superscript in a row differed significantly (P<0.05)
Table 6: Average serum-biochemical constituents, enzyme activities and mineral profile in growing Hariana heifers supplemented formaldehyde treated mustard cake
|Particulars||Treatment Group||Overall Mean||SEM||P- value|
|Total protein (g/dl)||5.81||6.03||6.07||5.85||5.94||0.05||0.20|
|Blood urea (mg/dl)||33.51||35.36||32.75||32.14||33.44||0.56||0.21|
Similar digestibility of DM, CP, CF and Acid detergent fibre (ADF) in all the four groups revealed that formaldehyde treatment of mustard oil cake had no adverse effect on nutrient intake and digestibility of nutrients. Similar results have earlier been reported by several workers on feeding of formaldehyde treated cakes to animals (Mathur et al., 1998; Devant et al., 2000; Chatterjee and Walli, 2003; Sahoo et al., 2006). From Table 4, it can be seen that average total DMI (kg/d), DM intake (kg/d) on percent body weight basis, metabolic body weight fortnightly (kgW0.75), feed conversion ratio (kg DMI/ kg weight gain) during the 120 days growth trial were similar in all the groups. These results corroborate with studies conducted by previous researchers, who also reported that protected protein by formaldehyde treatment did not influence DM intake (Tiwari and Yadava, 1990, 1994; Sahoo et al., 2006; Pratihar and Walli, 1995; Chatterjee and Walli, 2003), feed conversion ratio (Yadav and Chaudhary, 2006; Bugalia and Chaudhary, 2010) for different physiological functions. Similar to our findings, feeding of formaldehyde treated soyabean meal; sunflower seed meal and cotton seed meal did not lead to any improvement in feed efficiency, nutrient utilization, and performance of small ruminant’s performance (Abdullah and Awawdeh, 2004). Average live weight gain (fortnightly) and average daily gain were the highest in formaldehyde treated group than negative control group (P<0.05) but it was similar to positive control group. These findings are in agreement with Khan et al., (2000) who noticed the increased daily body weight gain in lambs fed with formaldehyde treated cottonseed meal. Improvement in average daily weight gain or growth rate in formaldehyde treated group and positive control group revealed that the formaldehyde treatment of mustard cake lowered the requirement of crude protein to meet the physiological demand. The higher growth rate in animals fed with formaldehyde treated cakes was also reported in previous studies (Cho et al., 1990; Tomilson et al., 1997; Chatterjee and Walli, 1998). Similarly, Divya et al.(2011) also noticed that daily body weight gain was improved due to high UDP ration.
The assimilation of minerals viz. Ca, Mg, Cu, Fe, Zn, Mn and Se absorption among the groups were statistically similar, while absorption of P was significant higher (P<0.01) in group II than that of other groups. The absorption (%) of P was higher for positive control group and the improvement may be due to the higher intake of P in positive control group which was due to high concentration of mineral mixture in group II that was 1.1% as compared to 0.8% in others. Research related to effect of formaldehyde treatment on mineral absorption is scanty. Park et al. (1999) reported that treatment with formaldehyde suppressed phytate degradation in formaldehyde treated soybean meal and rapeseed meal thus the absorption of P from oilseed meal is probably decreased by the treatment in ruminants. In the present study the presence of significant amount of inorganic phosphorus in the feed was not influenced by the formaldehyde treatment. Percent absorption of Ca, P, Fe and Zn were in agreement with Gupta et al. (2014). Se absorption was in agreement with Chaudhary et al. (2010) and Mahima et al. (2012).
No significant difference was found in serum creatinine (mg/dl), cholesterol (mg/dl), blood urea (mg/ dl), serum total protein, albumin, globulin, SGOT and SGPT activity among different groups of heifers (table 6). Divya et al. (2011) and Shelke et al. (2012) did not find any effect of UDP on serum cholesterol, total protein, albumin and globulin level in heifers and buffaloes. Tiwari and Yadava (1990, 1994); Srivastava and Mani (1995) and Maiga and Schingoethe (1997); Kumar et al. (2006), who also reported undegradable protein level had no effect on serum urea nitrogen. The data from Table 6 revealed that the serum P, Mg levels were statistically similar (P>0.05) among all four groups, while the serum Ca level increased significantly for all the groups as the trial progressed. Divya et al. (2011) found higher concentration of serum calcium content due to high UDP ration. Significant higher level of Zn and concentration was observed in positive control group (P<0.01). Serum Fe, Cu, Mn, Se levels were statistically similar in among all four groups at different intervals. Serum Fe, Zn and Se concentration are in agreement with Gadberry et al. (2003), but the Cu level reported in present study was slightly higher than these workers. Shinde et al. (2007) reported the similar levels of Se in buffalo calves. Shengfa et al. (2011) also reported the similar level of Se concentration in growing beef heifers.
From the findings of present study, it may concluded formaldehyde treated mustard cake had no adverse effect on nutrient intake and their digestibility. Incorporation of formaldehyde treated mustard oil cake in the concentrate mixture improved growth rate by 23.5 percent of Hariana cattle heifers in comparison to negative control (untreated) group. Thus, formaldehyde treatment of mustard cake is beneficial in growing heifers without producing any alteration in the mineral absorption and maintaining the normal values of the serum biochemical constituents, enzymes activities and mineral profile.
Conflict of Interests
The authors of the manuscript do not have a direct relation that might lead to a conflict of interests for any of the authors.
The authors wish to thank Dean, College of Veterinary Sciences and Animal Husbandry and Hon’ble Vice chancellor, Uttar Pradesh Pandit Deen Dayal Upadhayay Pashu Chikitsa Vigyan Vishvidhyalaya Evum Go-Anusandhan Sansthan (DUVASU), Mathura, India; for providing all the necessary support and facilities for conducting this study.