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Agriculture and Agricultural Science Procedia 11 (2016) 137 – 142

International Conference on Inventions & Innovations for Sustainable Agriculture 2016, ICIISA
2016

Effect of Eggshell as a Calcium Source of Breeder Cock Diet

on Semen Quality
Krittiya Lertchunhakiata,*, Pornpan Saenphooma, Manassanun Nopparatmaitreea,
Supawadee Chimthonga
a
Faculty of Animal Sciences and Agricultural Technology Silpakorn University, Phetchaburi 76120, Thailand

Abstract

The experiment was conducted to determine the effects of calcium source (coarse limestone, fine limestone and grounded eggshell)
in breeder cock diets on feed intake and semen quality. Fifteen Arbor Acres breeder cocks at 61 weeks of age were randomly
allotted into 3 treatments of 5 replications, in Randomized complete block design. T1 ( control group) ; coarse limestone: fine
limestone ( 50:50) , T2; coarse limestone: egg shell ( 50:50) and T3; egg shell ( 100) . Body weight were recorded weekly, feed
intake were recorded daily. Ejaculates were collected by abdominal massage method twice weekly to evaluate semen quality. The
results showed that body weight and feed intake were not significantly different ( P>0. 05) . Semen quality were evaluated with
sperm concentration, sperm motility, live sperm, abnormal sperm were not significantly different ( P>0. 05) . In conclusion,
grounded egg shell can be used as calcium source in breeder cock diet with normal feed intake and semen quality.
© 2016 Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
© 2016 The Authors. Published by Elsevier B.V.
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the Faculty of Animal Sciences and Agricultural Technology, Silpakorn University.
Peer-review under responsibility of the Faculty of Animal Sciences and Agricultural Technology, Silpakorn University
Keywords: eggshel; calcium source; breeder cocks

1. Introduction

Hen eggshell, a waste material from domestic sources such as hatcheries, poultry farms, fast food industries, egg
product factories, homes and restaurant, has been a serious matter as global awareness regarding organic waste
materials and pollution problems was increased (Than et al., 2012, King’Ori et al., 2011).

* Corresponding author. Tel.: +6-603-259-4038; fax: +6-603-259-4038

E-mail address: krittiya_2549@hotmail.com

2210-7843 © 2016 Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
Peer-review under responsibility of the Faculty of Animal Sciences and Agricultural Technology, Silpakorn University
doi:10.1016/j.aaspro.2016.12.023
138 Krittiya Lertchunhakiat et al. / Agriculture and Agricultural Science Procedia 11 (2016) 137 – 142

The management of agricultural wastes is indispensable and a crucial strategy in global waste management. For
sustainable development, wastes should be recycled, reused, and channeled towards the production of value added
products (Abdulrahman et al., 2014).
Chicken eggshells can be used as an alternative soil stabilizer like lime since they have the same chemical
composition. Although, the most common source of calcium for layer feeds is limestone. Due to the composition of
the egg shell is approximately 98.2, 0.9, 0.9% calcium carbonate, magnesium and phosphorus (phosphate),
respectively (Romanoff et al., 1949). Additionally, eggshell membrane consists of collagen as a component. Collagen
is a type of protein, fibrous in nature that connects and supports other bodily tissues, such as skin, bone, tendons,
muscles and cartilage. Eggshell membrane collagen is very low in autoimmune and allergic reactions as well as high
in bio-safety and is of similar characteristics to other mammalian collagen. Eggshells/shell membranes have multiple
uses in nutrition, medicine, construction and art works (King’ori, 2011). Calcium accounts for 56% of the motility-
stimulating activity of fowl spermatozoa and also fowl seminal plasma. Restores and stimulates the motility of fowl
spermatozoa at 40 °C in presence of calcium (Ashizawa and Wishart, 1987). Ca/P ratio of eggshell was ≥1.67
(Abdulrahman et al., 2014). Calcium recommended in cocks diet for attaining good semen quality could not be the
same as recommended for female (Khalil et al., 2012). NRC (1994) and Kappleman et al. (1982) concluded that there
were no differences in the reproductive performance of broiler breeder cockerels fed 0.5 to 7 g of calcium daily per
bird. While NRC (1994) and Norris et al. (1972) reported that diets containing 0.1 percent nonphytate phosphorus
were satisfactory for Single Comb White Leghorn (SCWL) cockerels. Bootwalla and Harms (1989) found that no
more than 110 mg of nonphytate phosphorus per bird daily were needed for maintaining reproductive capacity and
bone integrity in broiler breeder cockerels. However, calcium and phosphorus metabolism should be relate, due to low
calcium levels increase phosphorus excretion and low phosphorus levels increase calcium excretion. Eggshell calcium
is probably the best natural source of calcium and it is about 90% absorbable (Bee, 2011). It is a much better source
of calcium than limestone or coral sources. Hence, utilization of eggshell waste inverting to animal diet which would
transform the waste eggshells into a valuable item; giving financial benefits to the competitive egg processing industry
including environmental friendly.
The scope of current study was to investigate the effect of eggshell as calcium source in Arbor Acres breeder cocks
diets on body weight, feed intake and semen quality.

2. Materials and Methods

2.1 Experimental animal and diets

Eggshell ( produced by Ross308 Hens) waste from Chaveewan hatcheries, Sriracha district, Chonburi province in
Thailand were collected and transported to laboratory area at Silpakorn University. The eggshell was washed
throughly tap water. Then, it was dried in hot air oven at 95 ˚C for 4 h. The dried eggshell was grounded, and storage
at room temperature until dietary treatment preparation. The composition of the eggshell was analyzed by proximate
analysis (Table 1.)
The experimental diet was formulated according to the standards prescribed in NRC (1994). Fifteen Arbor Acers
breeder cocks at the age of 61 weeks were randomly allotted into 3 treatments of 5 replications, in Randomized
complete block design. The animals were raised by feeding diet containing different calcium sources as T1; control
group coarse limestone: fine limestone ( 50: 50) , T2; coarse limestone: egg shell ( 50: 50) and T3; egg shell ( 100).
follow Ingredients composition and nutrient levels of the experimental diets (Table 2.). The experiment was designed
and conducted for a period of three weeks during summer season (March - April 2015) at the Poultry Farm, Faculty
of Animal Science and Agricultural Technology, Silpakorn University. During the period of study, the average
minimum-maximum temperature and relative humidity recorded were 34.1-36.5°C and 68-73 percent, respectively.
 Krittiya Lertchunhakiat et al. / Agriculture and Agricultural Science Procedia 11 (2016) 137 – 142 139

Table 1. Chemical composition of grounded eggshell analyzed by proximate analysis.

Chemical composition Amount (%)
Dry matter 99.40
CP 2.14
Calcium 29.87
Total phosphorus 0.16
moisture 0.60

. Table 2. Ingredients composition and nutrient levels of the experimental diets.

Item Treatment 1 Treatment 2 Treatment 3
Ingredient (%)
Corn ground 49.15 49.15 49.00
Broken rice 10.53 10.53 10.50
Extruded soybean 3.01 3.01 3.00
Soybean meal 07.24 07.24 24
Palm oil 1.00 1.00 1.80
Salt 0.38 0.38 0.38
Premix 0.50 0.50 0.50
DL- Methionine 0.32 0.32 0.32
Coarse limestone 5.52 5.52 -
Fine limestone 5.52 - -
Eggshell - 5.52 10.8
Feed cost (Baht/kg) 11.62 11.67 11.95
Composition by
calculated
ME (kcal/kg) 2,800 2,800 2,800
CP (%) 18.00 18.00 18.00
Calcium (%) 8.3 8.3 8.3
Phosphorus (%) 38.0 38.0 38.0

2.2 Body weight and Feed intake

The Arbor acres cocks were serially tagged to represent treatments and replicates. The cocks were housed in
individually cage compartments in an open-sided building, fed standard breeders diet containing about 18% crude
protein (CP) and 2,800 kcal/g metabolizable energy and given fresh cool clean water each morning ad libitum. The
cocks were fed with the experimental diets for 3 weeks and then subjected to a semen collection twice weekly. The
cocks were weighed on weekly basis, while feed intakes were recorded daily.

2.3 Semen quality evaluation

The breeder cocks were caged individually and trained before start of experiment in order to stimulate erectile and
ejaculatory responses to the abdominal massage method (Burrows and Quinn, 1937; Lake, 1957) of semen collection.
The semen was collected at twice weekly interval among 6.30-7.30 A.M. to examine the following parameters viz;
volume, motility, concentration, live and dead and morphological defect rates of sperm were estimated using methods
described by Etchu and Egbunike (2002).
Semen volume from each of the cocks were measured with the use of a collection tube graduated in ml. For
evaluation of motility, one drop of the diluted semen was placed on the slide and covered with cover slip. The sperm
motility was estimated by microscopic observation ( ×400 magnification) . Sperm motility was expressed as the
percentage of motile spermatozoa with moderate to rapid progressive movement. A minimum of five microscopic
fields was assessed to evaluate sperm motility on at least 200 sperms for each sample. The semen concentration was
140 Krittiya Lertchunhakiat et al. / Agriculture and Agricultural Science Procedia 11 (2016) 137 – 142

measured using haemocytometer. Live, dead and abnormal sperm were counted and expressed as the percentage of
by live, dead and abnormal sperm using 5% eosin and 10% nigrosin stains. The field for examination of spermatozoa
was chosen at random and at least a total of 200 spermatozoa was counted.

2.4 Statistical analysis

Data were statistically analyzed according to SAS program ( SAS, 2004) using GLM Procedure. Analysis of
variance was carried out using the following model: Yij= μ+ i+ j+ ij; Where Yij= individual observation, μ= overall
mean, i =block effect, j=treatment effect, ij=error. Differences among treatments means were separated by Duncan's
multiple range test (Duncan, 1955).

3. Results and discussion

3.1 Live performances

From the present study, the body weight for each group was initially comparable and no difference was found up
to the end of the trial. Throughout the 3 weeks period, the average feed intake of the control group was 181.81±25.29
g/bird/day, which was not significantly (P>0.05) different from those in the groups fed 50% eggshell (187.04±23.10
g/bird/day) and 100% eggshell (192.95±27.65 g/bird/day). No effect of dietary treatment was found for body weight
and feed intake among three groups (Table 3. ). The results are in agreement with Gongruttananun (2011) reported
that ground, steriled eggshell can be mixed with other calcium source in diet of Rhode Island Red Roosters, which
were not affected to body weight and feed consumption. Additionally, the results found in laying hens in previous
studies (Froning and Bergquist, 1990; Scheideler, 1998). However, breeder cocks should be controlled body condition
score (1-5) not exceed 2.5 with high libido and fertility, BCS affect the performance of breeder animals.

Table 3. Body weight and feed intake of cross-bred cocks fed with different calcium sources (P>0.05).
Parameters Treatment 1 Treatment 2 Treatment 3
bodyweight (g) 4,696.97±170.91 4,620.00±186.48 5,030.00±239.57
feed intake (g/d) 181.81±25.29 187.04±23.10 192.95±27.65

3.2 Semen characteristics

The mean semen volume revealed no significant difference among treatment groups during the experimental
period (Table 4.). These results are similar to those obtained by Eltohamy and Takahara (1985), Neuman et al. (2002),
Zhai et al. (2007), Parizadian et al. (2011) and Khalil et al. (2012) who demonstrated no significant effect in semen
volume of calcium, carnitine and zinc supplemented cocks.
The mean sperm concentrations showed no significant difference among treatment groups according to
Gongruttananun (2011) revealed that there were not significantly different in sperm concentration among Rhode Island
Red breeder cocks fed with difference calcium sources (ground fine limestone, 50% eggshell and 100% eggshell).
Furthermore, Khalil et al. ( 2012) observed no significant effect on semen concentration in local chicken strains
receiving calcium supplemented (1, 2 and 3%) groups. However, numerical record of highest semen concentration was
noticed in treatment 2% calcium group. Prabhakar et al. (2014) confirmed that calcium 2 percent in cross-bred cock diet
improved the sperm concentration (P<0.01)
Sperm motility is one of the most important criteria of semen quality. From the present study revealed that the
percentage of sperm motility was no significant difference between treatment groups ( Table 4. ) , agree with
Gongruttananun (2011) reported that no significant differences were found in term of sperm motility among Rhode
Island Red breeder cocks fed with difference calcium sources (ground fine limestone, 50% eggshell and 100% eggshell).
Although, dietary calcium fed to Barred Plymouth Rock roosters enhances percentage of sperm motility reported by
Kanyinji and Maeda (2010). The previous studies indicated that calcium plays a key role in ensuring good sperm
motility. Insufficient calcium for breeder cock induce sperm lack the ability and energy to penetrate an egg.
 Krittiya Lertchunhakiat et al. / Agriculture and Agricultural Science Procedia 11 (2016) 137 – 142 141

Percentage of viability sperm revealed no significant differences between control and treatment groups throughout
experimental period (Table 4.), consistent with Gongruttananun (2011) noted no significant (P>0.05) effect in percentage
of viability sperm among Rhode Island Red breeder cocks fed with difference calcium sources (ground fine limestone,
50% eggshell and 100% eggshell).
From the current study, no significant difference were found in abnormal sperms parameter (Table 4.), agree with
Gongruttananun ( 2011) reported that no significant differences were found in term of sperm abnormality among
treatment groups. Although, Prabhakar et al. (2014) recommended that calcium 2 percent in cross-bred cock diet can
reduced abnormal (P<0.01) sperms.

Table 4. Physical characteristics and semen quality of cross-bred cocks fed with different calcium sources (P>0.05).
Parameters Treatment 1 Treatment 2 Treatment 3 P-value
Physical characteristics
color milky white milky white milky white -
odor normal normal normal -
pH 7 7 7 -
Mass movement score (0-5) 5 5 5 -
Semen quality
Semen volume (ml) 0.39±0.04 0.40±0.02 0.39±0.03 0.52
Semen Concentration (106 sperm) 157.64±22.01 190.80±46.28 147.88±26.64 0.40
Sperm motility (%) 76.27±2.61 80.74±1.76 72.63±4.57 0.10
Sperm viability (%) 82.70±3.10 82.31±2.39 77.37±2.34 0.39
Sperm abnormality (%) 21.27±2.74 22.25±2.99 20.76±2.17 0.71

4. Conclusion

Ground egg shell can be used as calcium source in breeder cock diet with normal feed intake and semen quality.
Additionally, inverting of egg shell to breeder cock diet as an alternative application to reduce the environmental
pollution problems.

Acknowledgements

This work was financially supported by Silpakorn University Phetchaburi Information Technology Campus
Creativity and Research Fund.

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