EFFECT OF CHELATED MINERALS SUPPLEMENTATION ON GROWTH AND REPRODUCTIVE PERFORMANCE OF MURRAH BUFFALO HEIFERS BY ANURADHA VERMA (2015V13M) Thesis submitted to Lala Lajpat Rai University of Veterinary and Animal Sciences in partial fulfillment of requirement for the degree of MASTER OF VETERINARY SCIENCE IN LIVESTOCK PRODUCTION MANAGEMENT DEPARTMENT OF LIVESTOCK PRODUCTION MANAGEMENT COLLEGE OF VETERINARY SCIENCES LALA LAJPAT RAI UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, HISAR-125004 (HARYANA) 2017
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This is to certify that this thesis entitled "EFFECT OF CHELATED MINERALS
SUPPLEMENTATION ON GROWTH AND REPRODUCTIVE PERFORMANCE OF
MURRAH BUFFALO HEIFERS" submitted for the degree of MASTER OF VETERINARY
SCIENCES in the subject of LIVESTOCK PRODUCTION MANAGEMENT to the LALA
LAJPAT RAI UNIVERSITY OF VETERINARY AND ANIMAL SCIENCES, HISAR is
a bonafide research work carried out by ANURADHA VERMA, Adm. No. 2015V13M
under my supervision and that no part of the thesis has been submitted for any other degree.
The assistance and help received during the course of investigation has been fully
acknowledged.
(Dr. S.K. CHIKKARA)Major advisor
Principal ScientistDeptt. of Livestock Production Management,
LUVAS, HISAR, HARYANA- 125004
CERTIFICATE-II
This is to certify that this thesis entitled "EFFECT OF CHELATED MINERALS
SUPPLEMENTATION ON GROWTH AND REPRODUCTIVE PERFORMANCE OF
MURRAH BUFFALO HEIFERS" submitted by ANURADHA VERMA, Adm. No.
2015V13M to the Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, in
partial fulfillment of the requirement for the degree of MASTER OF VETERINARY
SCIENCE in the subject of LIVESTOCK PRODUCTION MANAGEMENT has been
approved by the Student's Advisory Committee after an oral examination on the same.
EXTERNAL EXAMINER MAJOR ADVISOR
HEAD OF THE DEPARTMENT
DEAN, POST-GRADUATE STUDIES
ACKNOWLEDGEMENTS
First of all, I bow my head before my parents for their boundless blessings and support, whichaccompanied me in all endeavors.
It gives me immense pleasure to express my profound sense of gratitude to my esteemed majoradvisor, Dr. S.K. Chikkara, Principal scientist, Department of Livestock Production Management for hisvaluable guidance, close supervision, concrete suggestions, peerless encouragement, constructive criticismand sustained interest all through entire span of investigation and preparation of this manuscript. I amindeed greatly indebted and thankful to my co-major advisor respected Dr. Subhasish Sahu (AssistantProfessor, Deptt. of LPM), who sustained his incessant and exhilarating support to me.
It is my pleasure to express my sincere gratitude to the Dr. H.K.Gulati Professor & Head,Livestock Production Management, members of advisory committee, Dr. Sajjan Singh Sihag, (PrincipalScientist, Deptt. of ANN), Dr. S.P.Dahiya, (Professor, Deptt. of AGB), Dr.Ashok Kumar (Professor &Head ,Deptt. of VSR dean PGS Nominee and my previous major advisor Dr.S.S.Grewal for the interesttaken to give me the right perspective of my research and valuable guidance for my future ventures.
I undertake the privilege to express my deep sense of gratitude to Dr. Sandeep Gupta, (Scientist,Deptt. of VPB) for the help rendered by him in one form or other during the investigation.
Words can never express my gratitude to my family members whose endless love, affection anduntiring support and everlasting blessings bring me here. Next to God, I owe all the credit for successfulcompletion of my Master programme to my father Mr.Amar Singh Verma for his motivation, inspirationand supporting me in every adverse conditions and my mother Mrs.Banarasi Verma for her great love andsacrifices. With unbound affection I spread special fragrance of thanks to my fiance Mr.Arun Kumar forhis care and boundless affection,my sweet younger brother Mr.Ravi Kumar for his encouragement and everwilling help and my companion Ms. Jaena. This dream would not be materialized without their love,support and constant inspiration.
The valuable support rendered by my Seniors and Batch mates (Dr. Ravi, Dr.Deepak, Dr.vikas,Dr.Ramesh, Dr.Sachin, Dr.Madhur, Dr.Sujata, Dr.Sushma Dr. Neelima, Dr.Sweety) and VLDA Satish,LT Krishan and all farm labours in smooth conductance of my research work is deeply acknowledged.
Friendship needs no studied phrases, polished face or winking wiles. They are my friends, who attimes criticized, scolded and encouraged me to keep my determinacy to reach at proper decision.
I gratefully acknowledge Lala Lajpat Rai University of Veterinary and Animal Sciences forgranting funds to pursue the research work. I was blessed with a lot of friends and the moments I spenthere are the most memorable moments in my life.
Last but not least, I am indebted to those ‘dumb creatures’, the centre of my profession, theyprovided me an opportunity to serve them better.
Place: HisarDate: 29-7-2017 (ANURADHA VERMA)
CONTENTS
CHAPTER NO. DESCRIPTION PAGE NO.
I INTRODUCTION 1-3
II REVIEW OF LITERATURE 4-17
III MATERIALS AND METHODS 18-24
IV RESULTS 25-39
V DISCUSSION 40-44
VI SUMMARY AND CONCLUSION 45-48
BIBLIOGRAPHY i-vii
LIST OF TABLES
TableNo. Description
PageNo.
2.1 Effect of organic trace minerals supplementation in ruminants 7
2.2 Effect of organic trace minerals supplementation in non-ruminants 8
3.1 Distribution of experimental buffalo heifers in different dietary treatments 19
- Reduced no. of days in open- Increased conception rate- No change in milk yield and composition
Chester-Jones et al.,2013
Cows Cu, Zn, Mn - Increased colostrum immunoglobulins- Increased milk fat- No change in milk yield, protein and SCC- Lower calf mortality- Increased services per conception
Formigoni et al.,2011
Bulls Cu, Zn, Mn, Co - Increase in motile sperm(65.5 vs. 56.1%)- Increase in progressive sperm(47 vs. 38.4%)- Increase in sperms with rapid motility(62.3 vs. 52.8%)
Rowe et al., 2014
Rams Cu, Zn-methionine - Reduction in DMI- Increased ceruloplasmin and ALP- No effect on growth- Increased immunity
Gowda et al., 2014
Kids Cu-methionine - No effect on growth- No effect on nutrient intake and Digestibility- Increased Cu balance
Waghmare et al., 2014
Rams Cu, Zn –methionine - No effect on body weight- Reduced sperm motility- Reduced motile sperm count- No effect on semen volume- No change in nutrient intake and digestibility- Increased wool yield
Shinde et al., 2012
Ewes Cu, Zn-methionine
- Marginally lower DMI- No effect on nutrient digestibility and growth- Higher bioavailability- Reduced faecal mineral excretion
Pal et al., 2014
Kids Se-yeast - No effect on body weight gain- No change in carcass quality
Sethy et al., 2013
8
Table 2.2 : Effect of organic trace mineral supplementation in non-ruminants
Animal/bird
Organic mineral Observations Reference
Layers Cu, Mn,Zn proteinates
- Lower egg loss, higher thickness,and increased strength of the shell- No change in egg weight- No effect on feed intake, feed conversion,specific weight, and Haugh unit of eggs
Stefanello et al.,2014
Poultry Zn proteinate - No effect on tibia and liver mineral Content- Increased immune response- No change in DMI & FCR
Mandal et al., 2011
Poultry Cr propionate - No effect on intake and weight gain- Increased milk fat- Lowered serum corticosterone level in heatstressed poultry-Recoup in blood glucose level was better
Rajalekshmi et al.,2012
Poultry Cu, Zn, Mn, Fe - DMI decreased at 50% level- Increased FCR- Increased tibia mineral concentration- Similar glutathione peroxidase and ferricreducing ability in blood plasma
Rao et al., 2013
Poultry Mineral proteinates - Higher retention rate and bioavailability- Cu and Zn antagonism could be avoided- No effect on phytase- Less mineral wastage and decrease in Pollution
Ao and Pierce,2013
Quail Fe, Zn-methionine - No effect on growth- No effect on nutrient intake and FCR- Increased bioavailability
Sannamani et al.,2013
Pig Organic Zn - No effect on growth- Increased antioxidant level
Hill et al., 2014
2.4 Benefits of using chelated minerals for livestock production
The positive effects of chelated minerals on animal performance appear mainly due
to higher bioavailability as compared to inorganic sources. There are several studies in
different animal species with different sources of different mineral elements, which have
revealed notable differences in the bioavailability of organic and inorganic minerals. Studies
suggest that binding of Cu, Zn, Fe and Mn with amino acids and peptides can enhance the
bioavailability of these trace minerals, thereby leading to improved milk production, growth,
reproduction and general health status in livestock (Senthilkumar et al., 2015). The benefit
of using chelated minerals has been found to improve reproduction, health, soundness and
growth for livestock. A chelated mineral for cattle has focused on the pregnant cattle and the
young growing calves for improved immunity (less disease or sickness), reproductive
performance (shorter days open, higher conception rates, or less embryonic loss), and herd
health. Mares receiving trace mineral supplementation in a combined inorganic/chelated
product had a tendency for a reduction in the number of cycles bred and in the number of
services per mare. However, there was no effect in conception rate (Ott and Asquith, 1994).
9
It is well known that providing adequate levels of trace minerals is required for proper
immune function. Because of the increased availability of chelated products it is thought
that their use will enhance immune function (Vandergrift, 1993).
2.4.1. Body weight gain
In many of the experiments, growth performance was better in the animals
supplemented with organic minerals. Zn-methionine supplementation improved growth rates
and body weights in lambs (Garg et al.,2008). Similarly, organic sources of Se improved
growth performance in guinea pigs (Chaudhary et al., 2010) and lambs (Kumar et al.,
2009).however, no or very less improvement was seen in lambs (Kumar et al., 2009b)
supplemented with organic Se in comparison with inorganic Se. Calves supplemented with
organic trace mineral sources had a greater final weight and average body weight gain than
calves supplemented with similar levels of trace minerals from inorganic sources (Kegley et
al., 2012).
Organic trace mineral supplementation had no effect on growth performance of the
animals (Rajalekshmi et al., 2012; Shinde et al., 2012; Waghmare et al., 2014). Research
shows that organic mineral sources are more bioavailable. However production responses to
supplementation have been variable. Function of minerals are structural, physiological,
catalytic and regulatory. They are indirectly involved in the growth of animal.
Johnson et al. (1988) conducted five 28-day trials using a total of 773 newly received
calves to examine the effect of zinc methionine on health and performance. Calves were fed a
control diet or the control diet supplemented with zinc methionine to supply 360 mg
Each figure is an average of five values ± standard error
Figure 4.2: Average daily body weight gain (g) at monthly interval
4.1.4 Body weight parameter
The average initial body weight of heifers were 327.00, 331.40 and 325.80 kg, while
final body weights at 120th days of experiment were 389.80, 398.00 and 387.60 kg, in
treatment groups T1, T2 and T3, respectively. Overall, average daily body weight gain under
three treatments during the whole experimental period was 523.33, 555.00 and 515.00 g/d, in
T1, T2 and T3 groups, respectively The results of the study revealed that total weight gain and
gain per day did not differ significantly (P>0.05) in heifers fed ration supplemented with
chelated minerals as compared to inorganic mineral mixture.
28
Table 4.4: The mean values of body weight gain in buffalo heifers under differentdietary treatments
AttributesTreatments
T1 T2 T3
Initial body weight (kg) 327.00±16.10 331.40±14.83 325.80±13.70
Final body weight (Kg) 389.80±16.74 398.00±13.97 387.60±14.30
Total body weight gain (Kg) 62.80±1.36 66.60±1.57 61.80±2.35
Body weight Gain/day (g) 523.33±11.30 555.00±13.07 515.00±19.62Each figure is an average of five values ± standard error
Figure 4.3: total body weight gain (kg) during the trial
4.2 Effect of supplementation of chelated minerals on different body measurements
Average values of body length, height, heart girth and abdominal girth of buffalo
heifers under different treatments have been presented in Table 4.5, 4.6, 4.7, and 4.8,
respectively.
At the end of experiment the mean values of body length and height of heifers were
133.60, 137.20, 135.40 cm and 128.40, 129.80, 126.80 cm in treatment groups T1, T2 and T3,
respectively (Table 4.5 & 4.6). Average body length and height did not differ significantly
(P>0.05) between the treatment groups.
Similarly, the total gain in heart girth and abdominal girth were also non significant
(P>0.05) among treatment groups as shown in table 4.7 and 4.8.
29
Table 4.5: Body length of buffalo heifers during experimental period
AttributesTreatments
T1 T2 T3
Initial body length (cm) 124.40±1.97 127.20±1.88 126.20±1.72Final body length (cm) 133.60±1.86 137.20±1.93 135.40±1.50Total body length gain (cm) 9.20±0.49 10.00±0.70 9.20±1.36Each figure is an average of five values ± standard error
Table 4.6: Body height of buffalo heifers during experimental period
AttributesTreatments
T1 T2 T3
Initial body height (cm) 122.80±1.69 122.60±2.87 120.20±1.99
Final body height (cm) 128.40±1.89 129.80±2.48 126.80±2.04
Total body height gain (cm) 5.60±0.75 7.20±0.49 6.60±0.60Each figure is an average of five values ± standard error
Table 4.7: Heart girth of buffalo heifers during experimental period
AttributesTreatments
T1 T2 T3
Initial heart girth ( cm) 174.40±174.40 173.80±2.54 174.40±1.99Final heart girth (cm) 185.40±2.77 185.00±2.61 185.00±2.55Total heart girth gain (cm) 11.00±1.05 11.20±0.86 10.60±0.75Each figure is an average of five values ± standard error
Table 4.8: Abdominal girth of buffalo heifers during experimental period
Final abdominal girth (cm) 197.40±2.16 197.80±3.25 196.80±3.40
Total Abdominal girth gain(cm) 11.40±0.68 12.80±1.02 11.20±0.97Each figure is an average of five values ± standard error
4.3 Dry matter intake (DMI)
4.3.1 Dry matter intake (kg/day)
The mean values of total dry matter intake per day (kg/d) during the experimental
period have been presented in Table 4.9 and fig. 4.4. It was observed that there was a linear
increase in DMI during progressive growth period of heifers under different dietary
treatments except at the end due to the commencement of summer, which results in reduced
dry matter intake in all the treatment groups. The dry matter intake values did not differ
significantly (P>0.05) throughout the study period.
30
Table 4.9: Average dry matter intake (kg/day) of experimental buffalo heifers atfortnightly intervals
DaysTreatments
T1 T2 T3
0 Day 7.97±0.08 7.72±0.55 7.05±0.6315 Day 7.98±0.13 7.84±0.40 7.56±0.3830 Day 8.64±0.17 8.52±0.30 8.31±0.0945 Day 8.40±0.15 8.38±0.17 8.36±0.1160 Day 8.26±0.19 8.38±0.17 8.37±0.2775 Day 8.26±0.11 8.43±0.14 8.37±0.2090 Day 8.49±0.11 8.70±0.29 8.64±0.08
105 Day 7.91±0.09 7.93±0.18 7.58±0.08120 Day 7.07±0.17 7.12±0.10 6.94±0.10
Each figure is an average of five values ± standard error
Figure 4.4: Average Dry matter intake (kg/day) at fortnight interval
4.3.2 Dry matter intake (kg) per 100 kg body weight:
The mean values of dry matter intake per 100 kg body weight in heifers of all the
three treatment groups have been presented in Table 4.10.
The statistical analysis of the data showed no significant difference of DMI per 100
kg body weight in heifers under different dietary treatment during the whole experimental
period.
31
Table 4.10:Average dry matter intake (kg) per 100 kg body weight of experimentalbuffalo heifers at monthly intervals
4.4.2 Feed conversion efficiency (FCE)For the estimation of FCE, body weight gain (g) per kg of DM intake was calculated.
The average data for body weight gain (g) per kg of DM intake at monthly interval have beenpresented in Table 4.12.
After thirty days of feeding trial FCE values were 5.72, 6.06 and 5.79 in T1, T2 and T3,
respectively. The result did not show any significant (P>0.05) difference in FCE valuesamong the different treatment groups. The FCE values at the end of experiment, for T1, T2 andT3 treatments were 5.40, 5.63 and 5.58 respectively.
32
Table 4.12:Feed conversion efficiency (BW gain g/kg DMI) of experimental buffaloheifers at monthly interval
Each figure is an average of five values ± standard error
4.5 Effect of supplementing chelated minerals on nutrient digestibility, nutrient intakeand nutritive value
4.5.1 Nutrient DigestibilityDigestibility coefficients of dry matter (DM), crude protein (CP), crude fiber (CF),
ether extract (EE), nitrogen free extract (NFE), neutral detergent fiber (NDF) and aciddetergent fiber (ADF) in heifers fed ration supplemented with different levels of chelatedmineral have been presented in Table 4.13.
The average digestibility coefficients of dry matter were 65.27, 66.98 and 64.81 percent in dietary treatment groups T1, T2 and T3, respectively. It was observed that dry matterdigestibility did not differ significantly. The digestibility coefficients of crude protein were67.25, 69.95 and 66.09 per cent for T1, T2 and T3 groups, respectively. The statistical analysisrevealed that crude protein digestibility did not differ significantly(P>0.05). The mean values ofCF digestibility coefficients were 58.57, 59.09 and 57.94 per cent in experimental heifers oftreatment groups T1, T2 and T3, respectively. Digestibility coefficients of NFE were 70.31, 72.12and 70.24 for T1, T2 and T3 groups, respectively. Statistical analysis suggested that digestibilitycoefficient of crude fiber and NFE did not differ significantly (P>0.05). Ether extractdigestibility coefficients were 69.10, 69.93 and 68.89 for T1, T2 and T3 groups. The ADFdigestibility coefficients were 48.12, 48.89 and 47.60 for T1, T2 and T3 respectively whereasNDF digestibility of T1,T2 and T3 groups were found to be 55.35, 56.41 and 53.33 respectively.Table 4.13:Effect of supplementing chelated minerals on nutrient digestibility of buffalo
heifers
ParticularsTreatments
T1 T2 T3
DM 65.27±2.77 66.98±1.20 64.81±4.90
CP 67.25±0.96 69.95±1.92 66.09±1.96
CF 58.57±1.27 59.09±0.86 57.94±1.14
EE 69.10±0.57 69.93±0.61 68.89±0.62
NDF 55.35±1.04 56.41 ±0.72 53.33 ±1.11
ADF 48.12±0.60 48.89±0.62 47.60±0.79
NFE 70.30±0.66 71.12±0.48 70.24±0.38Each figure is an average of five values ± standard error
33
4.5.2 Nutrient intake
The dry matter intake during digestion trial were 7.28, 7.35 and 7.00 kg/d for T1, T2
and T3 groups respectively (Table 4.14). Present results suggested that the dry matter intake
was almost similar in all group of animals whether T1 (control) or T2 (50% replacement of
inorganic mineral mixture with chelated mineral) or supplemented with 100% chelated
minerals (T3). The dry matter intake per 100 kg BW (kg/d) and per kg metabolic BW
(g/kgW0.75) were found statistically similar (P>0.05) in all dietary supplemental groups.
Crude protein intake (CPI) and digestible crude protein intake (DCPI) during
digestion trial were 758.28 g, 759.80 g, 756.12 g and 509.64 g , 531.33 g, 500.01 g in T1, T2
and T3 groups, respectively (Table 4.14). Similarly digestible crude protein intake was
statistically similar (P>0.05) in heifers of different dietary treatment groups.
Table 4.14:Effect of dietary supplementation of chelated minerals on plane of nutritionduring digestibility trial
120 0.82a ±0.02 0.85b±0.02 0.92c±0.02Each figure is an average of five values ± standard errorThe mean values in a row with different superscripts differ significantly between the treatments (P<0.05)
4.7.4 Zinc concentration (mg/l)
The serum zinc concentration (mg/l) of experimental animals at monthly interval has
been presented in Table 4.19. The value ranges from 0.64 to 1.08 (mg/l) in control group (T1),
0.66 to 1.20 (mg/l) in T2 and 0.65 to 1.30 (mg/l) in T3. Data revealed that on or after 30 days
of experiment, serum zinc concentration was significantly (P<0.05) higher in treatment group
T3 than T1 and T2. Compared to day zero of experiment in all the groups serum zinc
concentration were increased as feeding trial advanced and it was highest on 120th day of trial.
The serum zinc concentration values then followed the similar trend during rest of the
experimental period.
Table 4.19:Serum zinc concentration (mg/l) of experimental buffalo heifers at monthlyintervals
DaysTreatments
T1 T2 T3
0 0.64±0.02 0.66±0.03 0.65±0.0230 0.72a±0.02 0.77a±0.01 0.84b±0.0260 0.82a ±0.01 0.86a ±0.01 0.99 b±0.0290 0.93 a ±0.01 0.98 a ±0.02 1.18 b ±0.02
120 1.08 a ±0.03 1.20 b ±0.02 1.30c ±0.02Each figure is an average of five values ± standard errorThe mean values in a row with different superscripts differ significantly between the treatments (P<0.05)
4.7.5 Manganese concentration (mg/l)The serum manganese concentration (mg/l) of experimental animals at monthly
interval has been presented in Table 4.20. The value ranges from 0.05 to 0.23 (mg/l) in
control group (T1), 0.06 to 0.24 (mg/l) in T2 and 0.06 to 0.25 (mg/l) in T3. Serum manganese
concentration on and after 60 days of experiment was significantly (P<0.05) higher in the
treatment group T3 than T1 but was comparable (P>0.05) with T2.. Compared to day zero of
experiment in all the groups serum manganese concentration were increased as feeding trial
advanced and it was highest on 120th day of trial. The serum manganese concentration values
then followed similar trend till the end of experiment.
120 0.23a±0.01 0.24ab±0.01 0.25b±0.00Each figure is an average of five values ± standard errorThe mean values in a row with different superscripts differ significantly between the treatments (P<0.05)
4.7.6 Iron concentration (mg/l)
The serum iron concentration (mg/l) of experimental animals at monthly interval has
been presented in Table 4.21. The value ranges from 0.93 to 1.30 (mg/l) in control group, 0.94
to 1.34 (mg/l) in T2 and 0.92 to 1.42 (mg/l) in T3. Serum iron concentration on and after 30
days of experiment was significantly (P<0.05) higher in the treatment group T3 than T1 and
T2. The serum iron concentration values then followed similar trend till the end of experiment.
Table 4.21:Serum iron concentration (mg/l) of experimental buffalo calves at monthlyintervals
DaysTreatments
T1 T2 T3
0 0.93±0.01 0.94±0.01 0.92±0.0130 1.08a±0.02 1.14b±0.01 1.19c±0.0160 1.17 a ±0.01 1.21 b ±0.01 1.25 c ±0.0190 1.22 a ±0.01 1.27 b ±0.01 1.34 c ±0.01
120 1.30 a ±0.01 1.34 b ± 0.01 1.42 c ±0.00Each figure is an average of five values ± standard error.The mean values in a row with different superscripts differ significantly between the treatments (P<0.05)
4.7.7 Cobalt concentration (mg/l)
The serum cobalt concentration (mg/l) of experimental animals at monthly interval
has been presented in Table 4.22. The value ranges from 0.003 to 0.005 (mg/l) in control
group, 0.003 to 0.005 (mg/l) in T2 and 0.003 to 0.007 (mg/l) in T3. Compared to day zero of
experiment in all the groups serum cobalt concentration were increased as feeding trial
advanced and it was highest on 120th day of trial. The results of the study revealed that serum
cobalt concentration throughout the experiment is significantly (P<0.05) higher in the
treatment group T3 than T1 and T2 but T1 and T2 did not differ significantly (P>0.05).
120 0.005a±0.000 0.005a±0.000 0.007b±0.000Each figure is an average of five values ± standard errorThe mean values in a row with different superscripts differ significantly between the treatments (P<0.05)
4.8 Reproductive Performance
Age at first heat
Only four heifers out of 15 from all the treatment groups came in heat during the
whole experimental period. One heifer from T1, two heifers from T2 and one heifer from T3
came in heat. Age at first heat was 916 days for T1 heifer, 809 days and 826 days for T2 heifer,
and 883 days for T3 heifer. Days taken to come in heat after start of the feeding trial was 105
days (n=1) for T1, 85.5 days (n=2) for T2 and 130 days (n=1) for T3 group.
Age at first conception
Three heifers got conceived out of four came in heat. Age at first conception was 916
days for T1 heifer, 809 days for T2 heifer, and 883 days for T3 heifer. All the three animals
from T1, T2 and T3 groups conceived in their 1st heat and one animal in T2 group which was
in heat failed to conceive during the experimental period of 120 days.
Number of A.I per conception
Number of A.I required per conception for all the three animals were one for each
animal per treatment group.
4.9 Cost of feeding
The cost of feeding of various groups of buffalo heifers in terms of total cost and cost
per kg body weight gain under different treatments was calculated from the records of feed
and fodder consumption, considering the actual price of feed and fodder and total gain in
body weight during the whole experimental period and has been presented in Table 4.23.
The total cost of feeding for a period of 120 days was Rs. 7,302, 7,809 and 8,316 in
treatments T1, T2 and T3, respectively. The corresponding values for cost per kg body weight
gain were Rs. 165.59, 163.65 and 184.56 in treatments T1, T2 and T3, respectively, which is
lowest in T2 treatment i.e. it was most economical when expressed in term of cost per unit
weight gain.
39
Table 4.23:Total cost of feeding (Rs.) and cost of feeding per kg body weight gain ofbuffalo heifers under different treatments for experimental period
ratio, reproductive performance and cost of feeding is presented in following section.
5.1 Growth parameters
5.1.1 Body weight gain
The body weight gain (kg) of experimental buffalo heifers showed increasing trend in
all the three treatments upto day 90, from day 90 to 120 though the heifers gained body
weight but leisurely as compared to first three months. (Table 4.1). The average body weights
(kg) and metabolic body weights (kg W0.75) of all the experimental animals was observed to
be statistically non-significant (P > 0.05) among various treatment groups throughout the
experiment (Table 4.1 and 4.2). However, the body weight gain during period 90 to 120 days
was less, the possible reason could be due to the change of season from winter to summer.
The result of present investigation corroborated with the findings of Muehlenbein et al
(2008) who found no differences (P > 0.10) among control, inorganic and organic
supplemented treatment groups in cow BW or body condition scores at various times
throughout the study, similarly calf birth weights did not differ significantly (P > 0.10)
among treatment groups. Similar findings were observed by Gengelbach et al. (1994) who
found no significant difference in the BW change for first-calf heifers exposed to mineral
treatments. Ahola et al ( 2004) also found no differences in BW and body condition score of
multiparous beef cows fed control (no supplemental Cu, Zn, Mn), organic (50% organic and
50% inorganic) and inorganic(100% inorganic) . It is possible that different trace minerals
enhance growth of heifers by stimulating activities of enzymes involved in nutrient
utilization. Chelates have been of less concern in the ruminant animals, probably due to
rumen microbes and their involvement in digestion (Mohanta et al, 2014)
5.1.2 Average Daily weight gain
Average daily weight gain (g/day) by buffalo heifers under different treatments has
been presented in Table 4.3. The results showed that the average daily weight gain did not
differ significantly (P>0.05) between different dietary regimen of heifers throughout the
experimental period.
The mean values of daily body weight gain during the whole period of experiment
were 523.33, 555.00 and 515.00 g/d, in T1, T2 and T3 treatment groups, respectively. The
41
result of study indicated that average body weight gain did not differ significantly (P>0.05)
due to partial or complete supplementation of chelated minerals as compared to inorganic
mineral in the three treatment groups. The findings of present study are in agreement of
earlier study who reported that the source of mineral whether inorganic or organic
supplementation did not affect the cow body weight or body condition score (Olson et
al.,1999; Muehlenbein et al.,2001; Ahola et al.,2004). Mondal et al (2008) reported no
significant (P>0.05) difference in body weight gain and average daily gain in male crossbred
calves on supplementation of organic or inorganic minerals as per NRC(2000) requirement.
Similarly, Ahola (2005) also reported that neither ADG nor DMI were affected by either trace
mineral supplementation or source throughout the growing phase in calves.
However, in contrary to the present findings, Hong et al. (2002) observed an increase
of 5.5 – 11.4% in daily weight gain of beef steer when basal diet was supplemented with
chelated minerals as compared to inorganic minerals. Similarly Bhanderi et al (2010)
concluded that supplementation of MBOTMs at NRC requirement in male calves can improve
the body weight gain than that of inorganic trace minerals.
5.2 Body Measurement
The present results revealed that there was improvement in body length, height, heart
girth and abdominal girth of the buffalo heifers in all the three treatment groups during
progressive period of growth. The body parameters of buffalo heifers did not differ
significantly (P>0.05) due to supplementation of 50% or 100% organic minerals in place of
inorganic at any stage of experiment. Since the body weight did not differ significantly
(P>0.05) among treatment groups, so also body dimensions were comparable among
treatment groups. Non-significant difference in growth performance, body measurements due
to chelated minerals supplementation was observed in the current experiment, although this
has not been well addressed in the literature.
5.3 Dry matter intake (DMI)
5.3.1 Dry matter intake per day (DMI/d)
Mean daily dry matter intakes (kg/d) during the experimental period are given in
Table 4.9 and fig. 4.4. The mean values of dry matter intake increased during the progressive
period in heifers under all the dietary treatment groups except in the last month of experiment
because of change in season from winter to summer. The dry matter intake during the whole
experimental period did not differ significantly (P>0.05) among treatment groups. The
findings of the study are in agreement with earlier reports of Lamb et al.(2008) who reported
that daily dry matter intake of hay was similar among treatment groups. Similar findings
were also reported by Mondal et al.(2008), Smith et al.(1967). However, in contrary to the
present findings Mallaki et al. (2015) found that nutrient digestibility and dry matter intake
was higher in the lambs fed with the diet supplemented with chelated mineral.
42
5.4 Feed conversion Ratio
The results of the study revealed that feed conversion ratio as well as feed conversion
efficiency of heifers were improved with the progress of experiment except at the end of
experiment due to seasonal changes (winter to summer). FCR and FCE did not differ
significantly (P> 0.05) among T1, T2 and T3 treatment groups during various intervals of
experiment. The findings are in agreement with the study of Ahola et al.(2005) who reported
no differences in gain to feed ratio between control and supplemented cattle (P=0.70) or
between ORG and ING cattle (P=0.47). Nockels (1991) compared copper proteinate and
copper sulphate in steers and found that weight gain and feed efficiency were not affected by
copper source. However, in contrary to the present findings Mallaki et al. (2015) reported
improved FCR in lambs fed chelated minerals. Similarly Dey and Garg (2004) observed
significantly improved feed efficiency in weaned albino rats given organic Zn compared to
unsupplemented and ZnSO4 supplemented groups.
5.5 Nutrient digestibility (%) and Nutrient Intake
Statistical analysis suggested that digestibility coefficient of dry matter, crude protein,
ether extract, crude fiber, NFE, ADF and NDF did not differ significantly (P>0.05) among all
the three treatment groups during the study period. The results of the study deduced that
nutrients intake by experimental heifers in terms of crude protein(CP), digestible crude
protein (DCP) and total digestible nutrients intake (TDN) did not differ significantly
(P>0.05) during the study.
The present finding also corroborated with the finding of Mondal et al. (2008) who
concluded that digestibility of DM, CP, CF showed no significant difference (P>0.05) among
the minerals supplemented groups. Garg et.al.,(2008) reported that intake of dry matter (DM),
organic matter (OM), crude protein (CP), digestible CP and total digestible nutrients and
digestibility of DM, OM, CP, ether extract, neutral detergent fibre and hemicellulose were
comparable (P>0.05) among the three groups.
Our findings are not in agreement with the results of Zhang et al. (2013) who
concluded that 0.1% chelate Cu and Zn supplementation improved nutrient digestibility in
weanling pigs. Also the results of present study is not supported by Bhoot et al 1981 who
suggested that due to supplementation of chelated minerals in ruminant results in better
fermentability and utilization of organic matter.
5.6 Water Intake
Water intake did not differ significantly (P>0.05) between the treatment groups
during various stages of experimental trial.
Nutrient Intake
The results of the present study deduced that nutrients intake by experimental heifers
in terms of crude protein (CP) , digestible crude protein (DCP) and total digestible nutrients
43
intake (TDN) did not differ significantly (P>0.05). The present finding also corroborated with
the finding of Garg et al.(2008) the intake of dry matter (DM), organic matter (OM), crude
protein (CP), digestible CP and total digestible nutrients and digestibility of DM, OM, CP,
ether extract, neutral detergent fibre and hemicellulose were comparable (P>0.05) among the
three treatment groups.
5.7 Serum mineral concentration
The present study revealed that serum concentrations of Ca and P in the treatment
groups did not differ significantly (P>0.05) throughout the study period although, there was
an increasing trend of serum mineral concentration with the supplementation of minerals in
the diet of all the treatment groups .
It was further revealed that initially there was no significant effect (P>0.05) of
supplemental trace minerals on serum concentration of Cu, Fe, Mn ,Co and Zn. However, day
30 onwards serum copper, zinc and iron increased significantly (P<0.05) in the T3 groups fed
100% chelated mineral mixture. After day 60, T3 exhibited high (P<0.05) serum
concentrations of Manganese, as compared to control group T1 fed inorganic mineral mixture
and 120 day onwards cobalt concentration in serum became significantly (p<0.05) higher in
treatment group T3 than T1 group. Present study revealed serum minerals concentration of
heifers increased linearly with the increase of days due to mineral supplementation and the
effect was more pronounced in group supplemented with 100% chelated minerals that was in
treatment group T3 than T1 . T2 group supplemented with 50% organic and 50% inorganic
minerals showed significantly (P<0.05) higher serum minerals concentration as compared to
T1 but significantly (P>0.05) lower when compared with T3 group. Similar observation was
recorded by Bhanderi et al. (2010) who reported high (P<0.05) serum concentrations of Cu,
Zn and Mn in male calves fed MBOTMs as compared to control group fed inorganic
minerals. Mondal et al. (2008) also found serum mineral concentration of zinc, copper,
manganese and iron increased linearly (P<0.05) with the increase of days due to mineral
supplementation particularly in organic mineral (T3 and T4) supplemented group.
In another study Engle et al. (2000) who reported higher (P<0.05) serum Cu
concentration on 84 days due to CuSO4 and Cu-lysine supplementation. Kegley and Spears
(1994) also found enhanced serum Cu level from both sources of CuSO4 and Cu-lysine like the
present findings. However in contrary to the present findings Tambe et al. (1998) did not find
any improved trace mineral profile on chelated and non-chelated mineral supplementation in
calves.
Higher serum concentration of trace mineral with organic mineral supplementation
probably due to higher absorption and retention in tissue level (Boland, 2003). On the other
hand, the inorganic forms of trace minerals make the intestinal pH more alkaline leading to
precipitation due to formation of inorganic chelates in the gut. These may be the possible
44
reason for the higher concentration of macro and micro elements in the serum of the calves
supplemented with the organic form of minerals (Duz et al., 1996).
5.8 Reproductive parameters
The results of the present investigation revealed that reproductive performance in terms of age
at first heat, age at first conception and A.I per conception were not affected due to 100%
supplementation of chelated minerals in place of inorganic. However,the heifers fed 50%
chelated minerals replacing inorganic attained early pubertal age as compared to control
group. Similar findings reported by Yasui et al (2014) that organic Mn,Zn,Cu
supplementation in cows did not improved uterine health.
In contrary to our findings Stanton et al (2000) found that feeding beef cows with organic
mineral resulted in increased pregnancy rates during the subsequent breeding season.
Similarly, Manspeaker et al. (1987) fed 40 first-calf Holstein heifers a control diet or the
control diet plus an amino acid chelated mineral supplement. Mineral content of the control
diet was not reported. The amino acid chelated supplement supplied additional iron,
manganese, copper and zinc in addition to potassium and magnesium. The study was
conducted from approximately 30 days prepartum until heifers were confirmed pregnant by
rectal palpation. Incidence of periglandular fibrosis (a pathologic response in which
endometrial tissue does not regenerate properly after parturition) was significantly lower (10
vs. 58%)in heifers given chelated minerals. Although not statistically significant, ovarian
activity tended to be higher and embryonic mortality lower for heifers fed the chelated
mineral supplement.
5.9 Cost of feeding
The total cost of feeding for a period of 120 days was Rs.10,392 ,Rs.10,899 and Rs.
11,406 in treatments T1, T2 and T3, respectively has been presented in Table 4.19.
The corresponding values per kg body weight gain were Rs. 165.50, 163.65 and
184.56 in treatments T1, T2 and T3, respectively, which is lowest in T2 treatment i.e. it was
most economical when expressed in term of cost per unit weight gain. In the present
investigations the cost of feeding per unit live weight gain was lower in T2 treatment
supplemented with 50% inorganic and 50% chelated minerals. Though treatment T2 seems to
be better than other two groups, but statistically not significant.
45
CCHHAAPPTTEERR--VVII
SSUUMMMMAARRYY AANNDD CCOONNCCLLUUSSIIOONN
The present study was conducted from January to April month on fifteen apparently
healthy buffalo heifers, between 22 to 28 months of age. The animals were maintained at
Buffalo Farm, Department of Livestock Production Management, College of Veterinary
Sciences, Lala Lajpat Rai University of Veterinary and Animal Sciences (LUVAS), Hisar.
The experiment was conducted for a period of 120 days.
The fifteen buffalo heifers were randomly distributed into three treatment groups each
having five buffalo heifers in such a manner that average body weight and age of each
experimental group was statistically similar. The heifers of dietary treatment T1 (control
group) were fed with seasonal green fodder, wheat straw and conventional concentrate
mixture having 2% inorganic mineral mixture to meet out the nutrients requirements as per
feeding standards (Ranjhan, 1998) . While heifers under treatment groups T2 and T3 were fed
as like T1 but with concentrate mixture having 1% inorganic + 1% chelated minerals
(replacing 50% inorganic mineral mixture with chelated minerals in concentrate mixture of
control group) and 2% chelated minerals + no inorganic mineral mixture (replacing 100%
inorganic mineral mixture with chelated minerals in concentrate mixture of control group),
respectively. The amount of concentrate mixture was given to each group in such a way that
the experimental ration remains iso-proteinaceous. The quantity of different feeds given to
each group was adjusted at fortnightly intervals so that the overall DCP requirements of
buffalo heifers were met according to the change in body weight. Animals were given ad-lib
fresh water throughout the experimental period.
All the experimental animals were weighed before start of the experiment, and
thereafter at fortnightly interval using standard platform weighing balance. The weights were
recorded in the morning for two consecutive days, before feeding and watering of the animals.
The body weights were utilized to calculate metabolic body weight and growth rate or weight
gain per day at fortnightly intervals using standard formula. Daily feed intake during the
experimental period by individual buffalo heifers was determined on the basis of feeds and
fodder offered and weigh back. On the basis of feed and fodder consumption, dry matter
consumed per day, per 100 kg body weight and per kg of metabolic body weight by the
animals at fortnightly intervals were estimated. The DM consumed and body weight gain
were used to calculate the FCR i.e. the amount of dry matter intake in kilogram (Kg) required
for per Kg weight gain and FCE i.e. body weight gain (g) per kg of DM intake at fortnightly
intervals by animals during the experimental period for each treatment.
46
A digestion trial of 7 days was conducted at the end of experiment to know the effect
of treatments on digestibility of feed and fodder.
Blood samples were collected at the beginning of the experiments and thereafter, at
monthly interval before feeding and watering of the experimental animals. About ten milliliter
(ml) of blood was collected by jugular vein-puncture into a set of sterile plastic tubes without
anti-coagulant for serum minerals tests. Blood collected without anti-coagulant were
centrifuged at 2500 to 3000 rpm for 25 minutes and plasma was separated and used for
estimation of biochemical parameters in serum viz. plasma calcium (mg/dl) and phosphorus
(mg/dl) using kits procured from M/S Transasia Biomedical Limited with fully automated
Random Access Clinical Chemistry Analyzer (EM 200TM Erba Mannheim – Germany). The
serum samples were digested in digestion mixture consisting of nitric acid and perchloric acid
for estimation of minerals copper, zinc, manganese, iron and cobalt using by atomic
absorption spectrometer- model Pinaacle 900T, S/N PTAS13050201 of PerkinElmer
Company.
Reproductive parameters viz. Age at 1st heat, age at 1st conception and A.I per
conception were recorded during the experimental period to know the effect of chelated
minerals on the above indices.
The total cost of feeding by each buffalo heifer during the whole period was
calculated. The information about the prices of all feed ingredient including price of greens
prevailing at the time of purchase were obtained from the Department of Animal Nutrition of
the University. Cost of feeding per kg gain was also computed for different treatment groups.
Body weight and average metabolic body weight of experimental buffalo heifers at
fortnightly intervals under different treatment groups showed no significant (P>0.05)
difference during 120 days of experimental period. The average daily body weight gain
during the whole experimental period was 523.3, 555 and 515g/d in T1, T2 and T3 groups,
respectively The results of the study revealed that total weight gain and gain per day did not
differ significantly (P>0.05) among different dietary treatment groups throughout the
experimental trial.
Average increase in body length and height, total gain in heart girth and abdominal
girth were comparable (P>0.05) in different treatment groups.
Dry matter intake per 100 kg body weight and per kg metabolic body weight of
heifers at various intervals under different dietary treatments did not differ significantly
(P>0.05). The digestibility coefficients of crude fiber and NFE; and nutrients intake in terms
of CP, DCP and TDN did not differ significantly (P>0.05) among three treatment groups.
The feed conversion ratio as well as feed conversion efficiency of heifers were also
non- significant (P>0.05).
47
It was observed that the serum calcium, phosphorus concentration did not differ
significantly (P>0.05) among the treatment groups but copper, zinc, manganese, iron, cobalt
concentrations were significantly higher (P<0.05) in group T3 as compared to T1 and T2 .
The increased level of Cu, Zn, Mn, Co and Fe in the serum of the buffalo heifers
supplemented with chelated minerals might be due to the higher bio-availability of these
elements from chelated as compared to inorganic mineral mixture. Unlike the inorganic
form, chelated minerals do not participate in interaction and antagonism between the
minerals, leading to greater absorption from the gastro-intestinal tract.
It can be concluded that reproductive performance in terms of age at first heat, age at first
conception and conception were not affected due to 100% supplementation of chelated
minerals in place of inorganic. However, the heifers fed 50% chelated minerals replacing
inorganic attained early pubertal age as compared to control group.
The results of the present study revealed that mean value for feed cost per unit body
weight gain was marginally lower Rs. 163.65 for T2 as compared to T1 165.50 but the same
value for T3 was quite high owing to the high cost of chelated minerals.
Although the serum micro minerals concentration was found to be significantly
higher in T2 group (50% chelated and 50% inorganic minerals) and T3 group (100% chelated
minerals) but it is not able to justify the additional cost of chelated minerals. However,
numerically T2 group showed higher values for body weight, average daily weight gain, body
measurements and was also cost effective thus, more economical to the dairy farmers but
statistically non-significant. Thus, increasing the organic minerals in the diet of animals
simply increases the cost, which may not be required, as the inorganic mineral is meeting out
the need of animal. But 50% replacement of organic minerals with organic one can be
practised for marginal improvement in cost of production.
In our experiment there was no deficiency of any of the minerals in any of the
treatment group diet throughout the trial. Majority part of the present experiment was
conducted in the winter season, when sufficient green fodder was available to meet out the
bodily demand for minerals. The serum mineral concentration of the treatment groups for all
the animals were in normal level, and that was sufficient to meet out the growth requirement
of animals. The serum micro mineral concentration though found to be higher for organic
mineral fed group but was within normal range, which could not affect the growth, nutrient
utilization like inorganic fed group.
On the basis of the results obtained in the present study it may be inferred that
supplementation of different levels of chelated minerals (50% or 100%) in ration of Murrah
buffalo heifers of 22 to 28 months age during january to april month of experimental period
has no favorable effect on growth performance, body measurements, nutrient digestibility,
nutrient intake and nutritive value of diet along with feed conversion ratio and feed
48
conversion efficiency and reproductive performance.
FURTHER RESEARCH
• Define the optimal level of Chelated minerals added to the diet.
• Better define conditions where performance responses may be seen.
• More closely define the mode of action of metal proteinates to improve performance
in ruminant animals.
Plate 1: Buffalo heifers kept chained and fed individually
Plate 2: Body length measurement of buffalo heifer
Plate 1: Buffalo heifers kept chained and fed individually
Plate 2: Body length measurement of buffalo heifer
Plate 1: Buffalo heifers kept chained and fed individually
Plate 2: Body length measurement of buffalo heifer
Plate 3 : Heart girth measurement of buffalo heifer
Plate 4: Water being offered to buffalo heifers in measurable bucket separately
Plate 3 : Heart girth measurement of buffalo heifer
Plate 4: Water being offered to buffalo heifers in measurable bucket separately
Plate 3 : Heart girth measurement of buffalo heifer
Plate 4: Water being offered to buffalo heifers in measurable bucket separately
Plate 5: Blood collection from buffalo heifer
Plate 6: Weighing concentrate mixture Plate 7: Weighing green roughage
Plate 8: Weighing faecal matter during digestion trial of 7 days
Plate 9: Automated Random Access Clinical Chemistry Analyzer
i
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ABSTRACT
Title of Thesis : Effect of Chelated Minerals Supplementation onGrowth and Reproductive Performance of MurrahBuffalo Heifers
Full Name of the Degree Holder : Anuradha VermaAdmission Number : 2015V13MTitle of the Degree : Master of Veterinary Science in Livestock Production
ManagementName of Discipline : Livestock Production ManagementName and Address of Major Advisor : Dr.S.K.Chikkara,
Principal Scientist,Deptt. of, Livestock Production ManagementLUVAS, Hisar, Haryana (India).125004
Degree Awarding University LUVAS, HisarYear of Award of Degree : 2017Major Subject : Livestock Production ManagementTotal Number of Pages in Thesis : 48 + viiNumber of Words in Abstract : 337Key words: Chelated minerals, buffalo heifers, growth and reproductive performance.
The objective of the present study was to analyze the effect of chelated minerals supplementation on
growth and reproductive performance of Murrah buffalo heifers. Fifteen apparently healthy heifers (22 to 28
months age) were selected randomly and stratified into three treatment groups each having five animals basing on
their average body weight and age. In treatment T1 (control), animals were fed with seasonal green fodder, wheat
straw and conventional concentrate mixture having 2% mineral mixture while for treatment T2 and T3, animals
were fed similar to T1 but, inorganic mineral in concentrate mixture was replaced with 50% and 100% chelated
mineral, respectively. Feeding trial was conducted for a period of 120 days. Body weight of experimental buffalo
heifers at fortnightly intervals under different treatment showed no significant difference (P>0.05) throughout the
experimental period. Metabolic body weight, average daily gain, body measurements (body length, height, heart
girth and abdominal girth) were also followed the same trend. However, each parameter was marginally higher for
T2 group but statistically remained non-significant. Macro minerals (Calcium and Phosphorus) profile in blood
serum was non-significant among the treatment groups. However, serum micro minerals ( Cu,Zn,Mn,Fe and Co)
was found to be significantly (P<0.05) higher for T2 and T3 groups as compared to T1. Digestibility coefficients of