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Effect of Semen Diluents, Dilution Rates, and Storage Periods on Spermatozoa

Motility of Different Varieties of Guinea Fowl

Article  in  Journal of Veterinary Science and Technology · September 2016

DOI: 10.4172/2157-7579.1000378

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DOI: 10.4172/2157-7579.1000378

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ISSN: 2157-7579

Research Article Open Access

Effect of Semen Diluents, Dilution Rates, and Storage Periods on

Spermatozoa Motility of Different Varieties of Guinea Fowl
Keerthy AJ1, Omprakash AV2, Churchil RR3 and Hudson GH4*
1
Directorate of Animal Husbandry, Kerala, India
2
Poultry Research Station, Madhavaram Milk Colony, Chennai, Tamil Nadu, India
3
Institutional Livestock Farm Complex, Veterinary College and Research Institute, Orathanadu, Thanjavur, Tamil Nadu, India
4
Department of Poultry Science, Madras Veterinary College, Chennai, Tamil Nadu, India

Abstract
Twelve number of healthy male guinea fowls from each variety of pearl, white, white breasted and lavender were
selected based on the phenotypoic characters and trained for semen collection by abdominal massage technique. Pooled
semen from each variety, were diluted with the Lakes Semen Extender, Modified Beltsville Poultry Semen Extender, and
Beltsville Poultry Semen Extender each in the ratio of 1:3, 1:4 and 1:5. The spermatozoa motility was assessed at 0, 1,
2, 4 and 6 hours of storage. Maximum per cent motility of 83.33, 86.67 and 79.19 were observed in the semen of pearl,
white breasted and lavender varieties of guinea fowl diluted Beltsville Poultry Semen Extender in the ration 1:4 at 6 h of
storage. Maximum spermatozoa motility with white guinea fowl variety at maximum storage period was observed with the
semen diluted (1:3) with Beltsville Poultry Semen Extender (81.67%). The current study will be of immense useful for the
selection of semen diluents for better fertility through artificial insemination in guinea fowls.

Keywords: Spermatozoa motility; Guinea fowl; Fertility high temperature in the study area (13.1623°N, 80.2433°E) was was
between 22oC and 31oC. The birds used for the study were housed in
Introduction individual cages and maintained under standard feeding and manage
mental conditions.
Guinea fowls are hardy, semi wild poultry that are yet to be
genetically improved for commercial meat and egg production. The Materials and Methods
fertility in guinea fowls is low which forms the major constraint for its
genetic improvement and commercial exploitation. In addition, sexing Initially 12 healthy, mature male guinea fowls aged eight months
problems [1], seasonality of breeding [2-4], monogamous behavior were selected based on their phenotypic characters from each variety
[5] may also contribute to lower fertility rates. Artificial insemination of guinea fowl namely the pearl, white, white breasted and lavender.
is the novel tool for improving the fertility in domestic animals and They were housed in individual cages providing a floor space of 1 sq.
poultry. The technology can be successful only with superior semen ft. per bird. Breeder ration containing 17% crude protein, 2700 kcal of
quality. Spermatozoa motility is the important parameter that Metabolisable Energy, 3% calcium, 1% lysine and 0.50% methionine
determines the movement of spermatozoa in the female oviduct. It was was provided at ad libitum with free access to drinking water. The
reported that the number of progressively motile sperm per ejaculate feathers around the vent region of the birds were clipped off and were
was the most consistent and reliable trait correlated with fertility [6]. trained for semen collection for a period of about one month.
Positive correlations between spermatozoa motility and fertility have Semen was collected during early hours of the day between twice
been reported by several authors in different species of poultry [7,8]. a week following abdominal massage technique [14]. Pooled semen
Poultry spermatozoa are fragile, and the semen quality of raw semen sample collected was then diluted with three different diluents namely
deteriorates within one hour of collection [9]. Although published the LSE [15], MBPSE [16] and BPSE [17]. The pH and osmolarity were
reports regarding the seminology of guinea fowls are less, the advantages measured and adjusted to standard levels using DALAL pH meter and
of using the semen diluents in other poultry species were reviewed. The OSMOMAT 030 cryoscopic osmometer respectively.
semen diluents are reported to be used to extend semen, maintain the
livability and fertilizing capacity in vitro, and maximize the number of Exactly 100 µL of raw semen was diluted with 200 µL, 300 µL
hens that can be inseminated [10]. Therefore the efficiency of different and 400 µL of each diluent in sterile eppendorf tubes to get the final
semen diluents in maintaining the spermatozoa motility of different dilution of 1:3, 1:4 and 1:5 respectively. The tubes were then stored at
varieties of Guinea Fowl namely the pearl, white, white breasted and 5°C and the spermatozoa motility were analyzed at zero, one, two, four
lavender, under short term storage was studied.
Three semen diluents namely the Lakes Semen Extender (LSE), *Corresponding author: Hudson GH, Department of Poultry Science, Madras
Modified Beltsville Poultry Semen Extender (MBPSE) and Beltsville Veterinary College, Chennai, Tamil Nadu, India, Tel: +91-994-437-2582 ; E-mail:
Poultry Semen Extender (BPSE) were used for the current study. hudson.bvsc@gmail.com

The spermatozoa motility of raw semen of pearl, lavender and white Received August 20, 2016; Accepted August 29, 2016; Published September
varieties was reported as 87.00 ± 4.40, 90.11 ± 3.70 and 84.34 ± 5.11 02, 2016
respectively [11]. The superiority of BPSE over many othersemen Citation: Keerthy AJ, Omprakash AV, Churchill RR, Hudson GH (2016) Effect of
diluents were also reported in different poultry species [12-14]. The Semen Diluents, Dilution Rates, and Storage Periods on Spermatozoa Motility of
Different Varieties of Guinea Fowl. J Vet Sci Technol 7: 378. doi: 10.4172/2157-
decrease in spermatozoa motility with increase in the dilution ratio 7579.1000378
and storage periods were reported with BPSE and IMV diluents in the
ratio 1:2 and 1:3 stored for up to six hours [13]. No works regarding Copyright: © 2016 Keerthy AJ, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
MBPSE with guinea fowl semen could be traced. The current study was unrestricted use, distribution, and reproduction in any medium, provided the
carried out during October- December months where the average daily original author and source are credited.

J Vet Sci Technol, an open access journal

ISSN: 2157-7579 Volume 7 • Issue 6 • 1000378
Citation: Keerthy AJ, Omprakash AV, Churchill RR, Hudson GH (2016) Effect of Semen Diluents, Dilution Rates, and Storage Periods on Spermatozoa
Motility of Different Varieties of Guinea Fowl. J Vet Sci Technol 7: 378. doi: 10.4172/2157-7579.1000378

Page 2 of 6

and six hours of storage. A drop of diluted semen with the aid of a diluted with LSE and MBPSE in 1:3 dilution rates with significant
micropipette was placed on a microscope slide, which was then covered (P ≤ 0.05) reduction after 4 h post-dilution. However there was no
with a glass cover slip and examined under 400 x magnifications in a significant (P ≥ 0.05) difference between storage periods in semen
light microscope. The motility determination was carried out by taking diluted with BPSE at 1:3 dilution rates. At 1:4 dilution rates, significant
into consideration subjective measurements based on the judgment of change in motility was observed in semen diluted with LSE (P ≤ 0.01)
individuals making the determination. The motility was expressed as and BPSE (P ≤ 0.05) but not with MBPSE. The reduction of motility
the percentage of cells that are motile under their own power [18]. became significant (P ≤ 0.05) at 2 and 4 h post-dilution in LSE and
MBPSE respectively. In case of 1:5 dilution rates, significant (P ≤ 0.01)
The statistical analysis of the data was carried out by one way ANOVA
reduction in motility was noticed only in semen diluted with LSE.
using Statistical Package for the Social Sciences 20.0 software [19].
Irrespective of the dilution rates, there observed significant
Results difference (P ≤ 0.05) between the extenders at 6 h of storage with BPSE
Effect of different semen extenders, dilution ratio and storage maintaining significantly (P ≤ 0.05) higher motility compared to other
periods on motility of pearl, white, white breated and lavender guinea two semen extenders. The different dilution rates with a particular
fowl spermatozoa at 5°C (Mean ± SE) were presented from Tables 1- 4 semen extender at any particular storage period did not have any
respectively. significant effect on motility.

Pearl variety White variety

The duration of storage had a significant (P ≤ 0.05) effect in semen The duration of storage had a significant effect in both semen

Treatment combinations (n=6) LSE MBPSE BPSE F value

1:3,0 hour 90.00b ± 4.66 88.33c ± 2.79 91.67 ± 2.79 0.265 NS
1:3,1 hour 82.50ab ± 6.92 85.00bc ± 2.24 88.33 ± 3.80 0.719 NS
1:3,2 hours 79.17 ± 6.11
ab
82.50 abc
± 2.50 86.67 ± 2.79 0.843 NS
1:3,4 hours 70.83a ± 6.11 80.00ab ± 2.24 83.33 ± 2.11 2.773 NS
1:3,6 hours 64.17Aa ± 6.38 74.17ABa ± 2.39 81.67B ± 2.79 4.471*
F value 3.669 *
3.464 *
1.880 NS

1:4,0 hour 90.00c ± 3.87 88.33 ± 4.01 93.33b ± 2.11 0.390 NS

1:4,1 hour 85.83bc ± 3.96 85.00 ± 4.28 90.00ab ± 1.83 0.520 NS
1:4,2 hours 81.67 ± 3.33
ab
80.83 ± 4.73 89.17 ± 2.01
ab
1.763 NS
1:4,4 hours 76.66ab ± 3.33 77.50 ± 4.79 86.67a ± 1.05 2.989 NS
1:4,6 hours 69.17Aa ± 2.39 75.00AB ± 5.16 83.33Ba ± 2.11 4.695*
F value 5.137** 1.796NS 3.596*
1:5,0 hour 87.50d ± 3.35 84.17 ± 3.01 91.67 ± 2.79 1.992 NS
1:5,1 hour 85.00cd ± 2.58 79.17 ± 4.17 90.00 ± 3.16 2.884 NS
1:5,2 hours 80.83bc ± 2.01 75.83 ± 4.55 87.50 ± 4.03 2.859 NS
1:5,4 hours 75.83 ± 1.54
ab
72.50 ± 5.13 83.33 ± 3.80 2.397 NS
1:5,6 hours 67.50Aa ± 1.12 70.00A ± 5.00 81.67B ± 4.60 3.867*
F value 10.198** 1.791 NS 1.157 NS
Ratio 0 hour 1 hour 2 hours 4 hours 6 hours

LSE ,1:3 90.00 ± 4.66 82.50 ± 6.92 79.17 ± 6.11 70.83 ± 6.11 64.17 ± 6.38

LSE ,1:4 90.00 ± 3.87 85.83 ± 3.96 81.67 ± 3.33 76.67 ± 3.33 69.17 ± 2.39

LSE ,1:5 87.50 ± 3.35 85.00 ± 2.58 80.83 ± 2.01 75.83 ± 1.54 67.50 ± 1.12

‘F’ value 0.396 NS

0.077 NS
0.061 NS
0.538 NS
0.348 NS

MBPSE,1:3 88.33 ± 2.79 85.00 ± 2.24 82.50 ± 2.50 80.00 ± 2.24 74.17 ± 2.39

MBPSE,1:4 88.33 ± 4.01 85.00 ± 4.28 80.83 ± 4.73 77.50 ± 4.79 75.00 ± 5.16

MBPSE,1:5 84.17 ± 3.01 79.17 ± 4.17 75.833 ± 4.55 72.50 ± 5.12 70.00 ± 5.00

‘F’ value 0.677 NS

0.911 NS
0.745 NS
0.802 NS
0.387 NS

BPSE,1:3 91.67 ± 2.79 88.33 ± 3.81 86.67 ± 2.79 83.33 ± 2.11 81.67 ± 2.79

BPSE,1:4 93.33 ± 2.11 90.00 ± 1.83 89.17 ± 2.01 86.67 ± 1.05 83.33 ± 2.11

BPSE,1:5 91.67 ± 2.79 90.00 ± 3.16 87.50 ± 4.03 83.33 ± 3.80 81.67 ± 4.60

‘F’ value 0.039 NS 0.114 NS 0.254 NS 0.347 NS 0.047 NS

Means bearing different superscripts in uppercase letter in a row and lowercase letter in a column differ significantly. **Highly significant (P≤0.01), * Significant (P≤0.05),
NS
- Not significant (P>0.05).
Table 1: Effect of different semen extenders- dilution ratio and storage periods on motility of pearl guinea fowl spermatozoa at 5°C (Mean ± SE).

J Vet Sci Technol, an open access journal

ISSN: 2157-7579 Volume 7 • Issue 6 • 1000378
Citation: Keerthy AJ, Omprakash AV, Churchill RR, Hudson GH (2016) Effect of Semen Diluents, Dilution Rates, and Storage Periods on Spermatozoa
Motility of Different Varieties of Guinea Fowl. J Vet Sci Technol 7: 378. doi: 10.4172/2157-7579.1000378

Page 3 of 6

Treatment combinations (n=6) LSE MBPSE BPSE F value

1:3,0 hour 80.00Ac ± 1.29 88.33Bb ± 2.79 91.67B ± 2.79 9.312**
1:3,1 hour 78.33Ac ± 1.67 86.67Bb ± 2.11 88.33B ± 3.81 9.646**
1:3,2 hours 74.17Abc ± 2.39 81.67Bab ± 2.47 86.67B ± 2.79 9.382**
1:3,4 hours 70.00 ± 2.58
Ab
77.50 ABab
± 4.96 83.33 ± 2.11
B
5.144*
1:3,6 hours 63.33Aa ± 2.11 70.83ABa ± 7.12 81.67B ± 2.79 4.430*
F value 10.868** 3.012* 1.880 NS
1:4,0 hour 83.33Ac ± 2.79 90.83Bb ± 2.39 90.83Bc ± 0.83 4.386*
1:4,1 hour 79.17 Abc
± 3.01 90.00 ± 2.58
Bb
88.33 Bbc
± 1.05 6.456**
1:4,2 hours 77.50Abc ± 3.60 85.00Bab ± 2.24 86.67Bb ± 1.05 4.047*
1:4,4 hours 72.50Aab ± 2.50 84.17Bab ± 2.71 81.67Ba ± 1.05 7.634**
1:4,6 hours 68.33Aa ± 2.79 79.17Ba ± 1.54 78.33Ba ± 1.67 8.530**
F value 4.370** 4.348** 18.369**
1:5,0 hour 82.50c ± 1.71 87.50 ± 4.23 92.50b ± 2.81 3.179 NS
1:5,1 hour 80.00 ± 1.83
bc
83.33 ± 3.58 88.33 ± 2.79
ab
2.556 NS
1:5,2 hours 75.83b ± 1.54 81.67 ± 4.22 85.83a ± 2.01 3.286 NS
1:5,4 hours 69.17Aa ± 2.39 78.33AB ± 4.01 83.33Ba ± 2.79 5.035*
1:5,6 hours 63.33 ± 2.47
a
70.00 ± 6.71 79.17 ± 3.01
a
3.372 NS
F value 14.293** 2.205 NS 3.861*
Ratio 0 hour 1 hour 2 hours 4 hours 6 hours
LSE,1:3 80.00 ± 1.29 78.33 ± 1.67 74.17 ± 2.39 70.00 ± 2.59 63.33 ± 2.11
LSE,1:4 83.33 ± 2.79 79.17 ± 3.01 77.50 ± 3.59 72.50 ± 2.50 68.33 ± 2.79
LSE,1:5 82.50 ± 1.71 80.00 ± 1.83 75.83 ± 1.54 69.17 ± 2.39 63.33 ± 2.47
‘F’ value 0.857 NS 0.162 NS 0.487 NS 0.490 NS 1.394 NS
MBPSE,1:3 88.33 ± 2.79 86.67 ± 2.11 81.67 ± 2.47 77.50 ± 4.96 70.83 ± 7.12
MBPSE,1:4 90.83 ± 2.39 90.00 ± 2.58 85.00 ± 2.24 84.17 ± 2.71 79.17 ± 1.54
MBPSE,1:5 87.50 ± 4.23 83.33 ± 3.58 81.67 ± 4.22 78.33 ± 4.01 70.00 ± 6.71
‘F’ value 0.241 NS 1.602 NS 0.360 NS 0.808 NS 0.757 NS
BPSE,1:3 92.50 ± 1.71 89.17 ± 1.54 87.50 ± 1.71 85.00 ± 1.83 81.67 ± 2.79
BPSE,1:4 90.83 ± 0.83 88.33 ± 1.05 86.67 ± 1.05 81.67 ± 1.05 78.33 ± 1.67
BPSE,1:5 92.50 ± 2.81 88.33 ± 2.79 85.83 ± 2.01 83.33 ± 2.79 79.17 ± 3.01
‘F’ value 0.742 NS 0.069 NS 0.255 NS 0.751 NS 0.534 NS
Means bearing different superscripts in uppercase letter in a row and lowercase letter in a column differ significantly, Highly significant (P≤0.01), Significant (P≤0.05),
** *

NS
-Not significant (P>0.05).
Table 2: Effect of different semen extenders - dilution ratio and storage periods on motility of white guinea fowl spermatozoa at 5°C (Mean ± SE).

diluted with LSE (P ≤ 0.01) and MBPSE (P ≤ 0.05) at 1:3 dilution from from 0 h to 6 h of storage in all the dilution rates under study.
rates. However there was no significant (P>0.05) difference in semen Further, irrespective of dilution rates, the semen diluted with BPSE had
diluted with BPSE at 1:3 dilution rate. Significant (P ≤ 0.01) reduction maintained superior spermatozoa motility at 6 h of storage compared
in motility observed in semen diluted with LSE, MBPSE and BPSE at to LSE and MBPSE diluents.Lavender
1:4 dilution rate from 0 to 6 hours of storage. The reduction in motility
The duration of storage had high significant (P ≤ 0.01) effect in all
became evident (P ≤ 0.05) at 4 h post-dilution in LSE and BPSE and 6
other combinations except for LSE in 1:3 dilution rates. Further BPSE
h post-dilution in MBPSE. At 1:5 dilution rate there was no significant
has maintained higher motility at 6 h storage period at all dilution rates
reduction in motility in semen diluted with MBPSE, however there was
compare to other semen extenders. There was significant difference
significant reduction in motility in semen diluted with LSE (P≤0.01) observed in motility for semen diluted with LSE, MBPSE and BPSE
and BPSE (P ≤ 0.05). Further semen diluted with BPSE at 1:3 and except for 1:4 at 4 h of storage period. The dilution rate alone showed
BPSE and MBPSE at 1:4 dilution rate maintained higher motility at 6 significant (P ≤ 0.05) difference in semen diluted with MBPSE at 0 and
h storage period. 6 h and BPSE at 1 and 2 h. The dilution rate alone showed highly significant
The superiority of BPSE over other diluents was much evident with (P ≤ 0.01) difference for MBPSE at 1 h and BPSE at 0 h of storage period.
1:3 and 1:4 dilutions, irrespective of the storage periods. Whereas, with 1:5 LSE was not influenced by dilution rate for different storage periods. After
dilution, significant difference in spermatozoa motility, was observed only dilution, the motility was progressively reduced in all semen extenders and
at 4 hours of storage. The dilution rate alone did not show any significant the reduction was significant (P ≤ 0.05) at 2 and 6 h post-dilution in 1:4
difference between storage periods for each semen extender. and 1:5 ratios with LSE, at 2, 4 and 6 h in all dilutions (1:3, 1:4 and 1:5).
The addition MBPSE at 1:3 ratios has resulted in time-related reduction
White breasted in spermatozoa motility at time intervals of post-dilution, 1 and 4 h at 1:4
The duration of storage had a significant (P≤0.05) effect on the ratio and 2 and 4 h at 1:5 dilution rates.
spermatozoa motility of the white breasted Guinea Fowl semen diluted
Discussion
with LSE at 1:3 dilution rate and MBPSE at 1:3 and 1:5 dilution rates
with reduction in motility from 0 h to 6 h of storage. With respect to Based on the dilution studies it was interpreted that among the
BPSE, highly significant (P≤0.01) reduction in motility was observed diluents, the BPSE has shown its superiority over other diluents

J Vet Sci Technol, an open access journal

ISSN: 2157-7579 Volume 7 • Issue 6 • 1000378
Citation: Keerthy AJ, Omprakash AV, Churchill RR, Hudson GH (2016) Effect of Semen Diluents, Dilution Rates, and Storage Periods on Spermatozoa
Motility of Different Varieties of Guinea Fowl. J Vet Sci Technol 7: 378. doi: 10.4172/2157-7579.1000378

Page 4 of 6

Treatment combinations (n=6) LSE MBPSE BPSE F value

1:3,0 hour 90.00Ac ± 4.08 87.50Ab ± 2.81 97.50Bb ± 1.12 5.935*
1:3,1 hour 85.00Abc ± 5.16 85.00Ab ± 3.87 95.83Bb ± 1.54 4.393*
1:3,2 hours 76.67Aab ± 6.01 84.17ABab ± 2.39 91.67Bb ± 1.05 4.768*
1:3,4 hours 71.67 Aab
± 5.87 79.17 ABab
± 2.39 87.50 ± 2.50
Ba
4.703*
1:3,6 hours 67.50Aa ± 5.74 75.00Aa ± 2.58 85.00Ba ± 1.83 6.424**
F value 3.830* 3.052* 8.667**
1:4,0 hour 87.50 ± 5.59 90.83 ± 4.55 98.33c ± 1.05 3.293 NS
1:4,1 hour 84.17 ± 7.24 85.83 ± 4.55 95.83 ± 0.83
b
2.489 NS
1:4,2 hours 79.17 ± 6.25 82.50 ± 4.79 92.50ab ± 1.12 2.941 NS
1:4,4 hours 75.00 ± 5.92 79.17 ± 5.23 88.33a ± 1.05 2.558 NS
1:4,6 hours 70.00 ± 5.92
A
75.83 AB
± 5.39 86.67 ± 2.11
Ba
3.837*
F value 1.735 NS 2.287 NS 13.847**
1:5,0 hour 85.83A ± 5.23 87.50Ac ± 3.82 96.67Bc ± 1.67 4.825*
1:5,1 hour 83.33 ± 5.73
A
83.33 Abc
± 4.01 94.17 Bbc
± 1.54 3.706*
1:5,2 hours 77.50 ± 5.59 77.50abc ± 5.29 90.00ab ± 0.00 3.442 NS
1:5,4 hours 70.83A ± 5.39 71.67Aab ± 4.60 85.83Ba ± 2.01 4.962*
1:5,6 hours 67.50 ± 5.74
A
68.33 Aa
± 4.94 83.33 ± 2.79
Ba
4.399*
F value 2.620 NS 3.639* 8.671**
Ratio 0 hour 1 hour 2 hours 4 hours 6 hours
LSE ,1:3 90.00 ± 4.08 85.00 ± 5.16 76.67 ± 6.01 71.67 ± 5.87 67.50 ± 5.74
LSE ,1:4 87.50 ± 5.59 84.17 ± 7.24 79.17 ± 6.25 75.00 ± 5.92 70.00 ± 5.92
LSE ,1:5 85.83 ± 5.23 83.33 ± 5.73 77.50 ± 5.59 70.83 ± 5.39 67.50 ± 5.74
‘F’ value 0.249 NS 0.043 NS 0.073 NS 0.195 NS 0.078 NS
MBPSE,1:3 87.50 ± 2.81 85.00 ± 3.87 84.17 ± 2.39 79.17 ± 2.39 75.00 ± 2.58
MBPSE,1:4 90.83 ± 4.55 85.83 ± 4.55 82.50 ± 4.79 79.17 ± 5.23 75.83 ± 5.39
MBPSE,1:5 87.50 ± 3.82 83.33 ± 4.02 77.50 ± 5.29 71.67 ± 4.60 68.33 ± 4.94
‘F’ value 0.863 NS 0.151 NS 0.634 NS 1.164 NS 0.940 NS
BPSE,1:3 97.50 ± 1.12 95.83 ± 1.54 91.67 ± 1.05 87.50 ± 2.50 85.00 ± 1.83
BPSE,1:4 98.33 ± 1.05 95.83 ± 0.83 92.50 ± 1.12 88.33 ± 1.05 86.67 ± 2.11
BPSE,1:5 96.67 ± 1.67 94.17 ± 1.54 90.00 ± 0.00 85.83 ± 2.01 83.33 ± 2.79
‘F’ value 0.273 NS 0.325 NS 2.059 NS 0.388 NS 0.524 NS
Means bearing different superscripts in uppercase letter in a row and lowercase letter in a column differ significantly, **
Highly significant (P≤0.01), Significant (P≤0.05),
*

NS
- Not significant (P>0.05).
Table 3: Effect of different semen extenders-dilution ratio and storage periods on motility of white breasted guinea fowl spermatozoa at 5°C (Mean ± SE).

in maintaining the motility of guinea fowl different varieties of abnormality [25]. The similar ionic changes that affect spermatozoa
spermatozoa under short term storage. The results obtained were in motility in guinea fowl semen needs to be further investigated
agreement with the previous studies in turkey semen [8,12]. In addition, with specific diluents under consideration. Identification of these
higher fertility percentage were reported in turkey semen diluted with attributes in future, may lead to efficient long term liquid storage of
BPSE extender stored for 24 hours [14] which may also be attributed to guinea fowl semen.
superior motility compared to IMV and MTGA diluents. However, the
superiority of CARI diluent over BPSE, LSE and normal saline for a 24 The dilution rate did not showed any significant difference between
hour storage period were also interpreted with higher fertility rates in storage periods irrespective of the extenders and the guinea fowl
guinea fowls [20]. The varying values obtained between the extenders varieties. However, the superior per spermatozoa motility at lower
may be due to different pH, composition of the diluents, metabolites dilution (1:2 and 1:3) were reported with BPSE and IMV diluents
released upon storage etc. [13]. The non-significant effects of dilution rates may be due to higher
dilution rates used in the current study.
Irrespective of the diluents and dilution rates,  the spermatozoa
motility showed a declining trend, as storage time progresses from Conclusion
0-6 hours. Similar results were also reported in Helmeted guinea fowl
semen preserved in powdered coconut water [21] and turkey semen The dilution studies revealed that, the semen diluted with BPSE
diluted (1:2) with poultry semen extender [22]. Complementary results (1:4) and MBPSE (1:4) or BPSE (1:3) in pearl and white variety of guinea
were also obtained in guinea fowl semen in CARI (24 h), BPSE (6 h) fowls respectively, and BPSE (1:4) in white breasted and lavender
and IMV (6 h) diluents in guinea fowl semen [13,23]. The reduction varieties, stored for 6 h is recommended, as the above combinations
in motility may be attributed to the release of sperm metabolites maintained superior motility, which may further yield better fertility
which alter the nutrient composition of the medium. Further, storage through artificial insemination in Guinea Fowls.
alters the osmolarity [24] of the medium, which leads to various Acknowledgements
sperm abnormality, which further reduces the progressive motility.
The authors are highly thankful to Dean, Madras Veterinary College, Chennai and
It was reported that storage of fowl semen in ringer’s solution causes Hon’ble Vice Chancellor, Tamil Nadu Veterinary and Animal Sciences University for
increased production of chloride ions which increases the spermatozoa providing all the necessary support and facilities for conducting this study.

J Vet Sci Technol, an open access journal

ISSN: 2157-7579 Volume 7 • Issue 6 • 1000378
Citation: Keerthy AJ, Omprakash AV, Churchill RR, Hudson GH (2016) Effect of Semen Diluents, Dilution Rates, and Storage Periods on Spermatozoa
Motility of Different Varieties of Guinea Fowl. J Vet Sci Technol 7: 378. doi: 10.4172/2157-7579.1000378

Page 5 of 6

Treatment combinations (n=6) LSE MBPSE BPSE F value

1:3,0 hour 82.50A ± 2.81 85.83ABd ± 2.01 90.83Be ± 0.83 4.751*
1:3,1 hour 78.33A ± 2.79 83.33Ad ± 1.05 88.33Bd ± 1.05 9.026**
1:3,2 hours 76.67A ± 3.58 78.33Ac ± 1.67 85.00Bc ± 0.00 4.453*
1:3,4 hours 74.17 ± 3.96
A
73.33 ± 1.67
Ab
81.67 ± 1.05
Bb
3.752*
1:3,6 hours 70.00A ± 4.08 61.67Aa ± 1.67 76.67Ba ± 1.05 8.782**
F value 2.041NS 31.176** 34.855**
1:4,0 hour 86.67Ac ± 1.67 88.33Ad ± 1.05 93.33Bc ± 1.05 8.585**
1:4,1 hour 81.67 Abc
± 1.67 85.00 Bcd
± 0.00 90.00 Cb
± 0.00 24.222**
1:4,2 hours 79.17Ab ± 2.39 80.00Abc ± 1.29 87.50Bb ± 1.12 8.227**
1:4,4 hours 75.00ab ± 2.58 76.67b ± 3.33 82.50a ± 1.71 2.511 NS
1:4,6 hours 70.00Aa ± 2.58 70.00Aa ± 3.16 79.17Ba ± 2.01 4.447*
F value 8.832** 13.361** 18.861**
1:5,0 hour 85.83Bc ± 0.83 82.50Ad ± 1.12 88.33Bc ± 1.05 8.291**
1:5,1 hour 80.83 ± 0.83
Ab
80.83 Acd
± 0.83 86.67 Bbc
± 1.05 12.940**
1:5,2 hours 79.17Ab ± 0.83 78.33Ac ± 1.05 84.17Bb ± 0.83 13.058**
1:5,4 hours 73.33Aa ± 1.67 72.50Ab ± 1.12 80.83Ba ± 0.83 13.769**
1:5,6 hours 70.83 ± 2.39
Aa
66.67 ± 1.05
Aa
78.33 ± 1.05
Ba
13.283**
F value 18.195** 36.826** 17.199**
Ratio 0 hour 1 hour 2 hours 4 hours 6 hours
LSE,1:3 82.50 ± 2.81 78.33 ± 2.79 76.67 ± 3.58 74.17 ± 3.96 70.00 ± 4.08
LSE,1:4 86.67 ± 1.67 81.67 ± 1.67 79.17 ± 2.39 75.00 ± 2.58 70.00 ± 2.58
LSE,1:5 85.83 ± 0.83 80.83 ± 0.83 79.17 ± 0.83 73.33 ± 1.67 70.83 ± 2.39
‘F’ value 1.244 NS 0.794 NS 0.288 NS 0.104 NS 0.022 NS
MBPSE,1:3 85.83 ± 2.01
ab
83.33 ± 1.05
b
78.33 ± 1.67 73.33 ± 1.67 61.67a ± 1.67
MBPSE,1:4 88.33b ± 1.05 85.00b ± 0.00 80.00 ± 1.29 76.67 ± 3.33 70.00b ± 3.16
MBPSE,1:5 82.50a ± 1.12 80.83a ± 0.83 78.33 ± 1.05 72.50 ± 1.12 66.67ab ± 1.05
‘F’ value 4.106* 7.308** 0.535 NS 1.196 NS 3.910*
BPSE,1:3 90.83ab ± 0.83 88.33ab ± 1.05 85.00a ± 0.00 81.67 ± 1.05 76.67 ± 1.05
BPSE,1:4 93.33b ± 1.05 90.00b ± 0.00 87.50b ± 1.12 82.50 ± 1.71 79.17 ± 2.01
BPSE,1:5 88.33 ± 1.05
a
86.67 ± 1.05
a
84.17a ± 0.83 80.83 ± 0.83 78.33 ± 1.05
‘F’ value 6.502** 3.750* 4.839* 0.472 NS 0.819 NS
Means bearing different superscripts in uppercase letter in a row and lowercase letter in a column differ significantly, Highly significant (P≤0.01), Significant (P≤0.05),
** *

NS
- Not significant (P>0.05).
Table 4: Effect of different semen extenders-dilution ratio and storage periods on motility of lavender guinea fowl spermatozoa at 5°C (Mean ± SE).

References 10. Lake PE (1956) A retarding factor in the problem of fowl semen storage. In:
Proc. of 3rd International Congress on Animal Reproduction. 3: 104-106.
1. Abdul-Rahman II, Awumbila B, Jeffcoate IA, Robinson JE, Obese FY (2015)
Sexing in guinea fowls (Numida meleagris). Poult Sci 94: 311-318. 11. Mohan J, Sharma SK, Kolluri G, Singh RP, Tyagi JS, et al. (2016) Semen quality
characteristics and seasonality in different varieties of male Guinea fowl. Adv
2. Ayorinde KL (1989) Effect of semen dosage and insemination frequency on the Anim Vet Sci 4: 320-326.
fertility of local pearly Guinea Fowl in Nigeria. Trop Agric 66: 135-136.
12. Iaffaldano N, Rosato MP, Manchisi A, Centoducati G, Meluzzi A (2010)
3. Konlan SP, Avornyo FK, Karbo N, Sulleyman A (2011) Increasing guinea fowl Comparison of different extenders on the quality characteristics of turkey
eggs availability and hatchability in the dry season. J World’s Poult Res 1: 1-3. semen during storage. Ital J Anim Sci 4: 513-515.
4. Premavalli K (2013) Influence of strain, age and system of management on 13. Hudson GH (2015) Fertility Parameters through artificial insemination in
the productive and reproductive performance of Guinea Fowl. PhD Thesis Guinea Fowl. M.V.Sc Thesis submitted to the Tamil Nadu Veterinary and Animal
submitted to the Tamil Nadu Veterinary and Animal Sciences University, Sciences University, Chennai.
Chennai.
14. Burrows WH, Quinn JP (1937) The collection of spermatozoa from the domestic
5. Aire TA, Ayeni JSO, Olowo-Okorun MO (1983) Some aspects of the fowl and turkey. Poult Sci 16: 19-24.
reproductive biology of the guinea fowl (Numida meleagris galeata Pallas). The
15. Lake PE, Ravie O (1982) Effect on fertility of storing turkey semen for 24 hours
Helmet Guinea Fowl (Numida meleagris galeata Pallas) in Nigeria, Kainji Lake
at 10°C in fluids of different pH. Br Poult Sci 23: 41-47.
Research Institute, New Bussa, Nigeria, pp: 189-193.
16. Kumararaj R, Omprakash AV (1996) Effect of different diluents, cryoprotectants
6. Kammerer DM, Moreng RE, Muller HD, Hobbs HW (1972) Turkey semen
and equilibration period on freezing of poultry semen. In: Proceedings of XX
evaluation for fertility prediction. Poult Sci 51: 77-82.
World Poultry Congress. Delhi, India, pp: 555-568.
7. Wishart GJ, Palmer FH (1986) Correlation of the fertilizing ability of semen 17. Sexton TJ, Giesen AF (1982) Beltsville Poultry Semen Extender. 6. Holding
from individual male fowls with sperm motility and ATP content.  Br Poult Sci turkey semen for 6 hrs at 15°C. Poult Sci 61: 1202-1208.
27: 97-102.
18. Parker JE, McKenzie FF, Kempster HL (1942) Fertility in the male domestic
8. Venkatesh G (2005) Effect of pre- Insemination vaginal douching and different fowl. Research Bulletin 347: 1-50.
semen extenders on fertility and hatchability in turkeys. MVSc Thesis submitted
to the Tamil Nadu Veterinary and Animal Sciences University, Chennai. 19. Snedecor GW, Cochran WG (1994) Statistical methods. IX edition. Oxford and
IBH publishing & Co., Kolkata, India.
9. Carter RD, McCartney MG, Chamberlin VD, Wyne JW (1957) The effect of
storage time and temperature on fertilizing capacity of Turkey semen. Poult 20. Mohan J, Sharma SK, Kolluri G, Tyagi JS (2015) Selection of Diluent for Short
Sci 36: 618-621. Term Preservation of Guinea Fowl Semen. Asian J Anim Vet Adv 10: 360-364.

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Citation: Keerthy AJ, Omprakash AV, Churchill RR, Hudson GH (2016) Effect of Semen Diluents, Dilution Rates, and Storage Periods on Spermatozoa
Motility of Different Varieties of Guinea Fowl. J Vet Sci Technol 7: 378. doi: 10.4172/2157-7579.1000378

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21. Lavor CTB, Câmara SR, Bandeira BM, Salgueiro CDM, Castro EV, et al. (2012) Physico-Biochemical Characteristics and Fertility of Guinea Fowl Semen. Adv
Powdered coconut water as preservation medium of guinea fowl (Numida Anim Vet Sci 1: 65-68.
meleagris) sperm. In: Ciência Animal 22: 79-81.
24. Clark RN, Bakest MR, Ottinger MA (1984) Morphological changes in Chicken
22. Słowińska M, Liszewska E, Dietrich GJ, Ciereszko A (2012) Characterization of and Turkey spermatozoa incubated under various conditions. Poult Sci 63:
proacrosin/acrosin system after liquid storage and cryopreservation of turkey 801-805.
semen (Meleagris gallopavo). Theriogenology 78: 1065-1077.
25. Saeki Y (1960) Crooked-Necked Spermatozoa in Relation to Low Fertility in the
23. Mohan J, Khanday JM, Singh RP, Tyagi JS (2013) Effect of Storage on the Artificial Insemination of Fowl. Poult Sci 39: 1354-1360.

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