The genetic improvement of offspring quality and potential for the propagation of
hybrids germplasm by assisted semen delivery into the female reproductive tract is
achievable through artificial insemination (AI). In vitro,
spermatozoa motility of fresh semen decreases within hour of collection and
consequently reduces fertility potential [1].
The importance of extender in poultry is to increase volume and facilitate prolonged
handling, maintaining good sperm motility and viability through inhibition of
pathways detrimental to semen. During semen handling and storage, there is often the
problem of the formation of lipid peroxides causing impaired spermatozoa plasma
membrane, which culminate into decrease in semen motility [2]. Seminal plasma possesses an antioxidant system to mitigate
the consequences of reactive oxygen species (ROS) accumulation; this seems to
protect sperm cells. Unfortunately, this protective capacity is limited in
spermatozoa compared with somatic cells [3,4].There are reports that suggests that herbs and fruits with antioxidant enrichment
protect cells by mitigating oxidative damage [5-7]. Bayemi et al. [8] reported that fruits based extenders
performed better in improving sperm motility than egg yolk extenders. Balogun and
Jimoh [9] developed garlic extracts as a
supplement in rooster semen diluent with appreciable success, but this requires high
standardization and techniques which limits its use by local farmers. Eggs are
relatively more expensive than fruits, thus tropical fruits based extenders may be
more economically effective in AI practise in farming households. Imminently,
growing interests in a new generation of semen extenders based on the presence of
natural products to minimize the risk of contamination through fruits containing
natural antioxidants which are more acceptable than synthetic antioxidants [10].Watermelon is a good source of lycopene and citrulline, it also contains large
amounts of beta-carotene and Vitamin C [11].
The therapeutic effect of watermelon has been ascribed to its composition of a
myriad of antioxidant compounds; chiefly citrulline and lycopene [12]. Water melon based diluents has shown great
potential in rabbit semen extension and AI [13]. There is growing interests to utilize fruits and natural extracts
as diluents and constituents of semen extender. This study attempt to harness the
potentials of water melon fortification of dextrose saline on roosters semen
extension.
METHODS
Experimental animals and management
This research was undertaken with approval from the institutional ethics
committee with IACUC approval no:- FPA/EC/19/0062 and the National Institutes of
Health guide for the care and use of Laboratory animals were followed, and
appropriate measures were taken to minimize pain or discomfort on the
animals.Thirty five to forty weeks old breeder roosters (30) and breeder hens (168) were
obtained from a reputable breeder farm were used for this study. Ripe water
melon (Sugar baby variety) fruit was purchased, washed, peeled and the fruit
pulp blended. Juice extractor (Mikachi model No 1706) was used to clarify the
fruit juice and was designated as watermelonjuice (WMJ) and kept frozen at
4°C in disposable 5 mL sterile sample bottles. Dextrose saline (5%
dextrose in 0.9% normal saline; Unique pharmaceuticals, Nig.) was procured for
the study. The experimental design is an entirely randomized design, consisting
of seven treatments, 8 replicates and three birds per replicate.Prior to collection of the semen, 2 weeks training for 30 roosters was undertaken
and semen were harvested twice weekly and ejaculates evaluated using
in-vitro analysis. Semen collection was carried out
according to the modified collection procedure outlined by Balogun et al. [14]. All roosters were assessed for
fertility and only roosters of good fertility were used, care was taken to avoid
any contamination of semen with faeces.
Extension of semen with extenders and evaluation
Seven (7) treatments consist of different diluent formulated. Treatment 1:
undiluted semen (positive control), Treatment 2: dextrose saline + 10% WMJ
diluent, Treatment 3: dextrose saline + 20% WMJ diluent, Treatment 4: dextrose
saline + 30% WMJ diluent, Treatment 5: dextrose saline + 40% WMJ diluent,
Treatment 6: dextrose saline + 50% WMJ diluent, Treatment 7: dextrose saline +
0% WMJ diluent (negative control).The pooled semen were allotted as described earlier and diluted with respective
extender dose. Diluted samples at 1 : 2 (semen : diluents) was equilibrated by
mixing gently and semen assessment took place immediately. Semen qualitative and
oxidative assay were assessed for each treatment. The assessments were done
hourly for 3 hours.
Semen evaluation
Extended semen according to treatments were evaluated for sperm kinetics using
computer assisted sperm analyzer (SpermAnalyzeWin7 Xuzhon city, China, setting
of CASA as in 5th WHO manual, 51 sperm tracks, evaluated magnification
×10, image acquisition rate: number frames/s 60); percentage motility,
progressive motility, non- progressive motility, curvilinear velocity (um/s),
average path velocity (um/s), straight line velocity (um/s), linearity,
straightness, amplitude of lateral head (um), beat cross frequency (Hz), wobble,
While sperm concentration and livability using convention procedures. Sperm
concentration (duplicates per sample) were determined using Neubauer
haemocytometer (TH-100, HechtAssistant, Sondheim, Germany) and expressed as
spermatozoa × 108/ mL. Livability was done by placing a drop
of semen on a glass slide, one drop of eosin-nigrosin stain added and mixed
gently, then smeared on a slide, air-dried and viewed under the microscope at
magnification of × 400.
Oxidative status assay
The semen samples of all treatment obtained at zero and three hours were
centrifuged and seminal fluid obtained was assayed for lipid peroxidation and
total antioxidant activity using standard procedures as outlined by Jimoh et al.
[6]. The assay for seminal lipid
peroxidation involve the reaction mixture in a total volume of 3.0 mL contained
1.0 mL seminal plasma and 1.0 mL of trichloroacetic acid (TCA) (0.67%). All the
test tubes were placed in a boiling water bath for a period of 45 min. The tubes
were shifted to the ice bath and then centrifuged at 12,000 g for 10 min. The
amount of malondialdehyde (MDA) formed in each of the samples was assessed by
measuring the optical density of the supernatant at 532nm [15].Seminal total antioxidant capacity activities involve a reaction mixture
containing 0.5 mL of a (10 mmol/L) Na-Benzoate, 0.2 mL of
H2O2 (10 mmol/L), 0.49 mL of phosphate buffer (100
mmol/L, pH = 7.4) (prepared by mixing 19.5 mL of KH2PO4
(100 mmol/L) with 80.5 mL of Na2HPO4 (100 mmol/L), then
adjusted the pH to 7.4 and 0.2 mL of Fe-ethylenediaminetetraacetate (EDTA)
complex (2 mmol/L) (prepared freshly by mixing equal volumes of EDTA (2 mmol/L),
and ferrous ammonium sulfate (2 mmol/L), then left at 25°C for 60 min.
Ten microliters of the seminal plasma were added to the latter reactive mixture
and were incubated at 37°C for 60 min. Finally, 1 mL glacial acetic acid
(20 mmol/L) and 1 mL thiobarbituric acid (0.8% w/v in 100 mL of 50 mmol/L NaOH)
were added, and the absorbance at 532 nm was measured spectrophotometrically
after incubation at 100°C for 10 min. Total antioxidant capacity was
calculated according to the following formula:Where CUA (mmol/L); concentration of uric acid; K: absorbance of the control (K1
– K0); A: absorbance of the sample (A1 − A0); UA: absorbance of
uric acid solution (UA1 – UA0) [16].
Artificial insemination
The hens were randomly divided into 7 treatments, each comprising of 8 replicates
and 3 hens per replicate. The hen insemination took place immediately after the
assessment of semen quality. Each hen was everted and semen deposited into the
intra vagina of the hen through the use of tuberculin syringe in which tubular
glass rod is attached to its mouth with the aid of rubber cork. The hens were
inseminated twice weekly in the evening for eight weeks. And the insemination
doses were 0.01 mL per hen (2 × 108 spermatozoa). Eggs were
collected for 7 weeks after the first week of insemination.
Egg fertility assessment
Eggs were collected daily, stored and incubated weekly. Eggs were incubated in a
reputable commercial hatchery. The fertility percentage ([fertile eggs / total
eggs] × 100) was determined by candling on the 18th day of incubation.
Using the formulae below:At every 18th day of incubation, eggs were transferred from the setter to the
hatcher (sections of the incubator) for the last 7 days were incubation and
hatching took place. Hatching percentage was determined by hatching of fertile
eggs about 21 days of incubation. Numbers of chicks hatched were counted and
percentage hatchability calculated using the formular below:
Statistical analysis
The data obtained were subjected to descriptive statistics and one way analysis
of variance of GLM procedure of IBM SPSS 25 at p = 0.05.
Differences in mean value were separated using New Duncan’s multiple
range test.The statistical model is as follow:Where Yijk represents the value of spermatozoa kinetics, oxidative stability and
fertility measured in the ith diluted semen; μ is the overall mean for
each character; Bi is the fixed effect of ith rooster semen diluted watermelon
(i = T1 is undiluted semen (positive control), watermelon was incorporated into
dextrose saline at 0%, 10%, 20%, 30%, 40%, and 50%); and eijk is the random
residual effect.
RESULTS
Semen quality of watermelon fortified rooster semen ab initio
The semen quality of watermelon fortified rooster semen is shown in Table 1. The semen motility and viability
was statistically similar across the treatments. Progressive motility values was
significantly (p < 0.05) highest in T3, T4, T5, and T6
semen groups, while values of T7 and T2 semen groups were not significantly
(p > 0.05) different. The significantly
(p < 0.05) lowest semen progressive motility was
observed in T1. Non- progressive motility of semen in T1 was significantly
(p < 0.05) highest. The significantly
(p < 0.05) lowest semen non progressive motility was
obtained in semen on T3, T4, T5, and T6. The average path velocity, linearity
and straightness of diluted semen (T2–T7) were significantly
(p < 0.05) higher than undiluted semen (T1).
Amplitude of lateral head of semen was (p < 0.05) higher
in undiluted semen than in the diluted semen group (T2–T7). Wobble of
semen in T2 was the highest, T1, T3, T4, T5, T6, and T7 semen groups were
lowest.
Table 1.
Semen characteristics of watermelon extended rooster semen at 0
hour
Parameters
T1
T2
T3
T4
T5
T6
T7
Percentage motility (%)
93.30
90.43
90.38
89.79
90.40
90.11
92.18
Progressive motility (%)
55.05[d]
69.64[c]
83.68[a]
81.08[ab]
84.55[a]
83.55[a]
72.57[bc]
Non- progressive motility (%)
38.25[a]
20.80[b]
6.70[c]
8.71[c]
5.85[c]
6.57[c]
19.61[b]
Curvilinear velocity (VCL)
(μm/s)
15.00[b]
21.45[b]
27.29[a]
28.94[a]
28.32[a]
28.22[a]
22.43[ab]
Average path velocity (VAP)
(μm/s)
5.74[b]
19.45[a]
19.40[a]
20.25[a]
19.70[a]
19.72[a]
22.46[a]
Straight line velocity (VSL)
(μm/s)
7.30[b]
9.72[a]
8.04[ab]
8.30[a]
8.02[ab]
8.09[ab]
9.48[a]
Linearity (%)
30.18[b]
52.90[a]
53.30[a]
52.71[a]
51.95[a]
52.44[a]
54.40[a]
Straightness (%)
46.38[b]
61.84[a]
62.89[a]
61.96[a]
61.46[a]
61.87[a]
63.93[a]
Amplitude of lateral head (ALH)
(μm)
0.76[a]
0.42[c]
0.50[c]
0.50[c]
0.48[c]
0.49[c]
0.62[b]
Beat cross frequency (BCF) (Hz)
4.48[a]
3.77[b]
4.35[a]
4.70[a]
4.68[a]
4.58[a]
5.00[a]
Wobble (%)
64.44[b]
70.78[a]
54.33[b]
54.13[b]
52.99[b]
53.57[b]
55.54[b]
Viability (%)
93.30
90.43
90.38
89.79
90.40
90.11
92.18
Means in the same row with different superscripts are significantly
(p < 0.05) different.
Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.
Means in the same row with different superscripts are significantly
(p < 0.05) different.Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.
Semen quality of watermelon extended rooster semen for 1 hour
Semen quality of watermelon extended rooster semen for 1 hour is shown in Table 2. Progressive motility of undiluted
semen (T1) was not significantly (p > 0.05) different
from the diluted semen groups. The percentage motility was significantly
(p < 0.05) highest in T2 and the significantly
(p < 0.05) lowest values is obtained in T7.
Progressive motility of semen in T1, T3, and T4 are not significantly
(p > 0.05) different but were significantly
(p < 0.05) higher than T2. Similarly,
non-progressive motility of semen on T2 was significantly (p
< 0.05) higher than semen on T1, T3, T4, and T5. Curvilinear velocity,
average path velocity and viability of the extended semen were not significantly
(p > 0.05) affected by the treatments. Straight line
velocity of semen on T2 was significantly (p < 0.05)
higher than semen on T4. Linearity of semen on T2 was significantly
(p < 0.05) higher than T6, while semen linearity
values for T1, T3, T4, and T5 had the significantly (p <
0.05) lowest values. Straightness of semen across the treatment was
significantly (p < 0.05) highest in T2 and T7 and values
for T1 and T3 was significantly (p < 0.05) lowest.
Amplitude of lateral head in semen on T6 was significantly (p
< 0.05) higher than those on T2. And the significantly
(p < 0.05) lowest values were found in T3 and T5.
Beat cross frequency was statistically similar in T1 and T5 and were
significantly (p < 0.05) higher than T7. Wobble of semen
on T1 and T5 was significantly (p < 0.05) lower than T7
and the significantly (p < 0.05) highest value was
obtained in semen on T2.
Table 2.
Semen characteristics of watermelon extended rooster semen at 1
hour
Parameters
T1
T2
T3
T4
T5
T6
T7
Percentage motility (%)
90.50[abc]
91.16[a]
90.73[abc]
89.67[bc]
89.78[b]
91.01[ab]
89.60[c]
Progressive motility (%)
83.99[a]
80.87[b]
84.45[a]
83.84[a]
83.65[ab]
82.67[ab]
82.19[ab]
Non- progressive motility (%)
6.51[b]
10.30[a]
6.28[b]
5.83[b]
6.13[b]
8.34[ab]
7.42[ab]
Curvilinear velocity (VCL)
(μm/s)
18.25
17.82
17.82
17.15
18.15
18.34
16.73
Average path velocity (VAP)
(μm/s)
9.79
10.23
9.69
9.25
9.71
10.07
9.32
Straight line velocity (VSL)
(μm/s)
4.08[ab]
4.44[a]
4.06[ab]
3.92[b]
4.08[ab]
4.31[ab]
4.04[ab]
Linearity (%)
22.33[c]
24.89[a]
22.79[c]
22.87[c]
22.49[c]
23.98[b]
24.18[ab]
Straightness (%)
41.65[c]
43.34[a]
41.90[c]
42.38[bc]
42.02[bc]
42.79[ab]
43.36[a]
Amplitude of lateral head (ALH)
(μm)
0.49[bc]
0.51[ab]
0.48[c]
0.50[ab]
0.48[c]
0.52[a]
0.50[ab]
Beat cross frequency (BCF) (Hz)
4.66[a]
4.29[ab]
4.51[ab]
4.27[ab]
4.67[a]
4.57[ab]
4.08[b]
Wobble (%)
53.59[cd]
57.44[a]
54.41[cd]
53.96[cd]
53.53[d]
54.88[bc]
55.76[b]
Viability (%)
90.50
91.16
90.73
89.67
89.78
91.01
89.60
Means in the same row with different superscripts are significantly
(p < 0.05) different.
Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.
Means in the same row with different superscripts are significantly
(p < 0.05) different.Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.
Semen quality of watermelon extended rooster semen for 2 hours
Semen quality of watermelon extended rooster semen for 2 hours is shown in Table 3. Percentage motility in undiluted
semen is not significantly (p > 0.05) different from the
extended semen groups (T2–T7). Progressive motility of semen in T1, T2,
T3, T5, and T6 were not significantly (p > 0.05)
different but were significantly (p < 0.05) higher than
semen on T7 and T4. Non progressive motility of semen in T4 and T7 were
significantly higher than other semen treatment groups. Curvilinear velocity of
semen in T4 was significantly (p < 0.05) highest and the
significantly (p < 0.05) lowest was observed T2. Average
path velocity in semen on T4 was significantly (p <
0.05) highest across the treatment, undiluted semen samples had statistically
similar values as diluted semen samples in T5 and T7. Straight line velocity of
undiluted semen samples was not significantly (p > 0.05)
different from semen in T2, T3, T5, and T6 and were significantly
(p < 0.05) higher than semen obtained in T7, while
the significantly (p < 0.05) highest values were
obtained in T4. Linearity of undiluted semen is not significantly
(p > 0.05) different from semen I n T2, T3, T5, and
T6 but were significantly (p < 0.05) lower than semen
samples in T4 and T7. Straightness of undiluted samples were significantly
(p > 0.05) similar with samples in T2, T3, and T6
but were significantly (p < 0.05) lower than semen in
T4, T5, and T7. Semen samples in T4 had the significantly (p
< 0.05) highest amplitude of lateral head while the significantly
(p < 0.05) lowest values is obtained in semen in T3
and T5. Beat cross frequency of undiluted semen was not significantly
(p > 0.05) different from semen samples in T3, T4,
T5, and T6, but were significantly (p < 0.05) higher
than semen on T2 and T7. The wobble in semen on T4 and T7 was significantly
(p < 0.05) higher than semen on T1, T2, T3, T5, and
T6. The viability of semen across the treatments were not significantly
(p > 0.05) different.
Table 3.
Semen characteristics of watermelon extended rooster semen at 2
hour
Parameters
T1
T2
T3
T4
T5
T6
T7
Percentage motility (%)
89.97[ab]
90.30[ab]
89.53[ab]
89.89[ab]
90.74[a]
89.23[b]
89.72[ab]
Progressive motility (%)
83.66[a]
85.22[a]
83.70[a]
75.50[b]
83.62[a]
83.83[a]
77.84[b]
Non- progressive motility (%)
6.31[b]
5.09[b]
5.83[b]
14.39[a]
7.12[b]
5.40[b]
11.87[a]
Curvilinear velocity (VCL)
(μm/s)
18.52[b]
17.61[c]
18.14[b]
19.94[a]
18.18[b]
18.25[b]
18.48[b]
Average path velocity (VAP)
(μm/s)
10.05[bc]
9.47[e]
9.67[d]
11.25[a]
9.82[c]
9.65[d]
10.34[b]
Straight line velocity (VSL)
(μm/s)
4.15[c]
3.98[c]
4.06[c]
4.85[a]
4.18[c]
3.97[c]
4.48[b]
Linearity (%)
22.38[bc]
22.61[bc]
22.35[bc]
24.39[a]
22.99[b]
21.74[c]
24.21[a]
Straightness (%)
41.25[c]
42.04[bc]
41.95[bc]
43.15[a]
42.55[ab]
41.09[c]
43.27[a]
Amplitude of lateral head (ALH)
(μm)
0.50[b]
0.50[bc]
0.49[c]
0.55[a]
0.49[c]
0.50[bc]
0.50[b]
Beat cross frequency (BCF) (Hz)
4.71[a]
4.44[b]
4.57[ab]
4.75[a]
4.61[ab]
4.68[a]
4.43[b]
Wobble (%)
54.25[b]
53.77[b]
53.29[b]
56.39[a]
54.03[b]
52.88[b]
56.94[a]
Viability (%)
89.97
90.30
89.53
89.89
90.74
89.23
89.72
Means in the same row with different superscripts are significantly
(p < 0.05) different.
Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.
Means in the same row with different superscripts are significantly
(p < 0.05) different.Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.
Semen quality of watermelon extended rooster semen for 3 hours
Semen quality of watermelon extended rooster semen for 3 hours is shown in Table 4. The percentage motility of
undiluted semen was not significantly (p > 0.05)
different from watermelon fortified semen (T2, T3, T4, T5, and T6) and were
significantly (p < 0.05) higher than semen diluted with
only dextrose saline (T7). Progressive motility of undiluted semen was
significantly (p > 0.05) similar with semen in T2 and
were significantly (p < 0.05) higher than semen in T3,
T4, T6, and T7, while significantly (p < 0.05) lowest
values was obtained in T5.
Table 4.
Semen characteristics of watermelon extended rooster semen at 3
hour
Parameters
T1
T2
T3
T4
T5
T6
T7
Percentage motility (%)
90.37[a]
90.02[a]
91.47[a]
91.99[a]
92.84[a]
90.99[a]
80.74[b]
Progressive motility (%)
80.66[a]
83.55[a]
66.08[b]
62.99[b]
46.70[c]
68.51[b]
61.36[b]
Non- progressive motility (%)
9.71[c]
6.47[d]
25.41[b]
29.00[b]
46.15[a]
22.49[b]
19.37[ab]
Curvilinear velocity (VCL)
(μm/s)
18.20[c]
17.79[c]
20.89[b]
26.68[a]
26.96[a]
23.24[b]
10.19[d]
Average path velocity (VAP)
(μm/s)
10.11[c]
9.65[c]
13.51[b]
14.88[b]
18.13[a]
13.04[b]
8.98[c]
Straight line velocity (VSL)
(μm/s)
4.31[c]
4.07[c]
6.51[b]
6.44[b]
8.41[a]
5.81[b]
5.42[c]
Linearity (%)
23.69[c]
22.88[c]
30.94[b]
24.14[c]
35.76[b]
24.98[c]
58.03[a]
Straightness (%)
42.60[bc]
42.22[bc]
47.31[b]
43.27[bc]
38.99[c]
44.54[b]
60.12[a]
Amplitude of lateral head (ALH)
(μm)
0.57[d]
0.49[d]
0.66[c]
0.73[b]
0.84[a]
0.65[c]
0.39[e]
Beat cross frequency (BCF) (Hz)
4.28[bc]
4.61[bc]
3.94[c]
5.54[a]
4.44[bc]
5.05[ab]
1.46[d]
Wobble (%)
55.59[c]
54.20[c]
64.41[b]
55.78[c]
67.42[b]
56.13[c]
88.33[a]
Viability (%)
90.37
90.02
91.47
91.99
92.84
90.99
80.74
Means in the same row with different superscripts are significantly
(p < 0.05) different.
Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.
Means in the same row with different superscripts are significantly
(p < 0.05) different.Treatments are % watermelon in dextrose saline diluents: T1 is
undiluted semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5,
40%; T6, 50%; T7, 0%.Non-progressive motility of semen samples in T5 was significantly
(p < 0.05) highest and the lowest values was
obtained in T2. Curvilinear velocity of semen samples in T4 and T5 were
significantly (p < 0.05) highest and significantly
(p < 0.05) lowest value was obtained in semen on T7.
Average path velocity and straight line velocity of undiluted semen is not
significantly (p > 0.05) different from T2 and T7 wand
were significantly (p < 0.05) higher than T3, T4, and
T6. Linearity, wobble and Straightness of semen diluted with only dextrose
saline was significantly (p < 0.05) higher than
undiluted semen and watermelon fortified semen groups. Amplitude of lateral head
of semen diluted with dextrose saline was significantly (p
< 0.05) lowest and the significantly (p < 0.05)
highest values was obtained in semen on T5.
Oxidative status and fertility assessment of watermelon extended rooster
semen
The antioxidant activity of seminal plasma declines across all the treatment
groups with time from zero hours to 5 hours (Fig.
1). At zero hour, as watermelon inclusion in semen increased it led
to corresponding increase in seminal antioxidant activity. The lowest values
were found in undiluted semen and dextrose saline diluted semen groups. Lipid
peroxidation of seminal of undiluted semen and dextrose saline diluted groups
increase from 0 hours to 5 hours (Fig. 2).
This is contrary to the rate of lipid peroxidation in watermelon-dextrose
diluted semen, which declined from 0 hours to 5 hours. The fertility of hens
inseminated with watermelon extended semen is shown in Fig. 3. The percentage fertility was highest in T3 (81.01%)
and T4 (81.24%) with lowest value obtained in T7 (73.46%). Hatchability of eggs
set of hens inseminated with undiluted semen (71.46%) was lower than values for
hens inseminated with watermelon inclusive extended semen
(75.71%–80.39%). The hatchability of fertile eggs was highest in hens
inseminated with T4 (99.07%) and the lowest values was obtained in hens
inseminated with T7 (94.8).
Fig. 1.
Antioxidant activity of rooster semen in watermelon-dextrose based
diluent.
Treatments are % watermelon in dextrose saline diluents: T1 is undiluted
semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5, 40%; T6, 50%;
T7, 0% (negative control).
Fig. 2.
Lipid peroxidation of rooster semen in watermelon-dextrose based
diluent.
Treatments are % watermelon in dextrose saline diluents: T1 is undiluted
semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5, 40%; T6, 50%;
T7, 0% (negative control).
Fig. 3.
Fertility of hens inseminated with watermelon-dextrose extended
rooster semen.
Treatments are % watermelon in dextrose saline diluents: T1 is undiluted
semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5, 40%; T6, 50%;
T7, 0% (negative control).
Antioxidant activity of rooster semen in watermelon-dextrose based
diluent.
Treatments are % watermelon in dextrose saline diluents: T1 is undiluted
semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5, 40%; T6, 50%;
T7, 0% (negative control).
Lipid peroxidation of rooster semen in watermelon-dextrose based
diluent.
Treatments are % watermelon in dextrose saline diluents: T1 is undiluted
semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5, 40%; T6, 50%;
T7, 0% (negative control).
Fertility of hens inseminated with watermelon-dextrose extended
rooster semen.
Treatments are % watermelon in dextrose saline diluents: T1 is undiluted
semen (positive control); T2, 10%; T3, 20%; T4, 30%; T5, 40%; T6, 50%;
T7, 0% (negative control).
DISCUSSION
Seminal plasma has limited antioxidant capacity, thus the need for an extender with
strong antioxidant effect to maintain the viability and subsequent fertilizing
capacity. This study elucidated the importance of watermelon in enhancing viability
of rooster semen. From in vitro results watermelon in dextrose
saline in semen dilution enhanced progressive motility and low non progressive
motility, this is in congruent with a recent study that showed WMJ as a potent semen
diluent [13]. Although, Adebisi and Ewuola
[17] documented the potency of dextrose
saline as a superior semen diluent in poultry. The higher performance of watermelon
fortified semen can be attributed to the phytoconstituents of watermelon chiefly,
antioxidants such as vitamin C, lycopene and beta carotene. Consequently, study have
stated that the addition of vitamin C to an extender could possibly improve sperm
function by reducing cell damage through its continuous radical-scavenging action
[18]. For instance, alpha-tocopherol
which is one of the main antioxidants, was found to be abundant in spermatozoa
membrane that protects sperm from oxidative damage [19]. Similarly, lycopene in tomatojuice acts as an antioxidant that
protect spermatozoa from oxidative damage in an extender composition [1]. Tvrda et al. [20] suggested that lycopene supplementation to semen extender
acts as an effective motion-promoting and membrane protecting molecule, by
significantly improving spermatozoa motility, membrane integrity, and mitochondrial
activity during preservation. The results shows that watermelon addition to rooster
semen diluent enhance the antioxidant prowess of rooster semen and lower lipidperoxide generation. This shows that watermelon inclusion in dextrose enhance
scavenging ability of the ROS and free radical and consequently reduced lipid
peroxidation in rooster semen. This would account for the trend in semen kinetics
and motility in this study. Amedu & Idoko [21] also reported that watermelon (Citrullus lanatus)
promotes normal sperm morphology, concentration, motility and volume of rat semen.
C. lanatus thus possess health benefits as it contains lycopene
with rich antioxidant. The inclusion of watermelon in rooster semen performed better
than undiluted semen despite over 3 hours storage, this is similar to results from
Mangiagalli et al. [22] that reported
positive effects of lycopene supplementation to extender on fowl sperm survival
during liquid storage. Report that vitamin C is naturally present in seminal plasma
to scavenge and decrease numerous disruptive free radical processes, including lipid
peroxidation [23], show the enhancement
potential of watermelon bound antioxidant complex in enhancing semen quality of
rooster semen. Thus sperm kinetics properties of extended semen tend to be better
with watermelon inclusion. This is similar to claims of Zheng & Zhang [24] that antioxidative compounds are beneficial
for sperm viability and reduction of lipid peroxidative damage to sperm
membranes.In vivo result showed that watermelon in dextrose saline when used
as diluent for rooster semen enhanced fertility and hatchability of eggs than
undiluted semen and dextrose saline. This thus corroborates claim by Jimoh &
Ayedun [13] that WMJ is a potent diluent and
can be incorporated as extender constituent in the preservation of semen for optimal
performance. Finally, watermelon based diluents enhance reproductive output of
animals in assisted reproductive techniques which is in line with report of
Mangiagalli et al. [25] that lycopene
addition affects reproductive performance of multiparous rabbit does inseminated
with fresh semen.
CONCLUSION
The supplementation of watermelon in dextrose saline as rooster semen diluent
revealed its potential to enhance spermatozoa motility and sperm kinetics over 3
hours of observation. The hatchability and fertility of eggs obtained from hens
inseminated with watermelon-dextrose saline diluted semen were better than only
dextrose diluted semen. The optimal inclusion of 30%–50% watermelon in
dextrose saline diluent enhance semen kinetics, fertility and hatchability of eggs
obtained from inseminated hens and were better than result obtained with undiluted
semen. It can be recommended that future efforts should focus on natural/organic
products in extender composition to cryopreserve rooster semen to utilize the
potentials of watermelon revealed in this study.
Fig. 1.
Fig. 2.
Fig. 3.