Open Access

Effect of supplementation of feed with Flaxseed (Linumusitatisimum) oil on libido and semen quality of Nilli-Ravi buffalo bulls

  • Syed Mazhar Hussain Shah1,
  • Shujait Ali1,
  • Muhammad Zubair1Email author,
  • Huma Jamil1 and
  • Nazir Ahmad1
Journal of Animal Science and Technology201658:25

DOI: 10.1186/s40781-016-0107-3

Received: 4 March 2016

Accepted: 31 May 2016

Published: 1 July 2016

Abstract

Background

The current study was designed to investigate the effect of supplementation of Flaxseed (Linumusitatisimum) oil on libido and semen quality of Nilli-Ravi buffalo bulls.

Methods

In this study, 12 adult healthy bulls kept at the Semen Production Unit, Qadirabad district Sahiwal, were used. These bulls were divided into three equal groups, A, B and C. Group A was kept as control, while in groups B and C supplementation of feed was provided by using flaxseed oil @125 ml/day and 250 ml/day,respectively for 12 weeks. Two ejaculates per animal were collected at 0 day then 5th, 6th, 7th, 8th, 9th, 10th, 11th and 12th week of treatment. In this way a total 216 samples were taken, and each semen sample was evaluated for color, volume, mass activity, percent motility, sperm cell concentration per ml, percentage of live sperm, and plasma membrane integrity. Libido of bulls was also evaluated before every collection.

Results

Analysis of data revealed that these parameters were significantly (P < 0.01) increased in flax oil treated animals as compared to control.

Conclusion

It was concluded from the present study that flax seed oil has beneficial effects on reproductive health of buffalo bull.

Keywords

Semen Flax seed and breeding bulls

Background

Male contributes 50 % of the herd and importance of male in genetic improvement of livestock cannot be denied. Genetic potential of a male is commonly exploited through artificial insemination. Reproductive capacity of males is judged by sperm production and success of artificial insemination depends upon number and quality of sperms in the ejaculate [6]. Semen is collected from genetically superior animals. In addition to other factors, volume of semen, sperm concentration, motility and number of normal sperm have great importance in conception.

In Mammalian sperm, lipids especially n-3 fatty acids are dominantly present [24]. These fatty acids are involved in motility of sperm, integrity of sperm membrane and protection of sperm from cold shock [25]. Animals are unable to synthesize the n-3 and n-6 fatty acids in their body as they do not have sufficient amount of fatty acid denaturize enzymes. Therefore, these fatty acids need to be taken by dietary sources [31]. Previous studies have shown that those fatty acids which are present in diet are also a part of sperm in human [8] and in farm animals [16]. Moreover, ration having fatty acids in diet improves value of post thawed buffalo bull semen [1].

Docosahexaenoic acid has been shown to increases serum testosterone level and sperm motility in man [8], boar [26], goat [9] and sheep [30] and flaxseed is a source of docosahexaenoic acid. Omega -3 fatty acids are necessary for cell membrane integrity and fitness of brain and eyes [8]. Main component of flaxseed is lignin, which contains plant estrogen and antioxidants and has protective effect on sex hormone metabolism [2].

Little information is available with regard to the effect of dietary flax seed oil supplementation and fertility of buffalo bulls during heat stress . The objective of present study was to investigate the effect of flaxseed oil supplementation on libido and semen quality parameters of Nilli-Ravi buffalo bulls.

Methods

Experimental animals

A total of 12 adult Nilli-Ravi buffalo bulls with clinically normal reproductive tract kept at the Semen Production Unit Qadirabad district Sahiwal, Pakistan were used in this study. These bulls were routinely used for semen collection. Animals were randomly divided into 3 equal groups A, B and C. Group A was kept as control while, group B and C was supplemented flaxseed oil drenched orally with dose of 125 ml and 250 ml respectively. All these treatments were continued for consecutive 12 weeks. The study period was from mid April to mid July, 2015. Semen was collected by artificial vagina at day 0 then 5th, 6th, 7th, 8th, 9th, 10th, 11th and 12th week of treatment. Two ejaculates per animal were collected on each collection day.

Parameters examined

Libido of each bull was assessed by mating behavior of bulls. Mating behavior was videotaped using video camera and video recorder. By watching videotapes libido in terms of reaction time to first semen collection was recorded [17]. Each ejaculate was evaluated for color, volume, mass activity, percent motility, sperm concentration, live sperm, and plasma membrane integrity. To examine mass activity, a small drop of semen was placed on pre warmed glass slide with glass rod and examined under microscope. On the base of wave pattern, mass activity score from0 to 5 was assigned to each sample [27]. To examine motility of spermatozoa, a drop of semen was placed on glass slide and mixed with a drop of 2.9 % sodium citrate. Slide was examined under 200X magnification. Sperm concentration was directly determined using Accucell Bovine Photometer. Sperm membrane functional integrity was determined by hypo-osmotic swelling test (HOST) which is based on semi permeability of cell membrane. The hypo-osmotic solution (150 mosm/L) was prepared as described by [19]. Eosin-nigrosin staining was used to determine live perm percentage.

Statistical analysis

Mean values (± SE) of various parameters for bulls of three groups were calculated. The differences in group were compared by “Tukey test”. The data was analyzed using SPSS software.

Results

Analysis of variance showed that there is (P < 0.01) improvement of libido in treated groups as compared to control. Interaction between weeks and groups was also significant (Table 1). Analysis of variance of Table 2 revealed that there was the significant effect of weeks and treatment on semen volume (p˂0.01). Further analysis showed that semen volume for bulls of group B and C was (p˂0.01) higher than the control. However, difference between group B and C was non-significant. The effect of weeks and treatment on semen mass activity of treated groups was significant (p˂0.01) higher as in Table 3. Likewise, weekly analysis of variance revealed that semen mass activity for bulls of group B and C was significantly higher than the control. However, there was non-significant difference between group B and C.
Table 1

Mean values (±SE) of Libido collected from bulls of control and treatment groups on different weeks

Weeks of Treatments

Group

Over all Mean

A (Control)

B (Treatment 1)

C (Treatment 2)

0

7.50 ± 0.50a

7.25 ± 0.48ab

7.50 ± 0.29a

7.42 ± 0.23A

5

7.50 ± 0.65a

4.75 ± 0.25cde

4.25 ± 0.48e

5.50 ± 0.50B

6

6.75 ± 0.48a-d

5.00 ± 0.41b-e

4.75 ± 0.48cde

5.50 ± 0.36B

7

7.25 ± 0.48ab

4.50 ± 0.29de

4.25 ± 0.25e

5.33 ± 0.45B

8

7.50 ± 0.29a

4.50 ± 0.29de

4.25 ± 0.48e

5.42 ± 0.48B

9

7.00 ± 0.71abc

4.00 ± 0.41e

4.00 ± 0.41e

5.00 ± 0.51B

10

7.25 ± 0.48ab

4.25 ± 0.48e

4.00 ± 0.41e

5.17 ± 0.51B

11

6.75 ± 0.25a-d

4.00 ± 0.41e

4.00 ± 0.41e

4.92 ± 0.43B

12

7.00 ± 0.41abc

4.00 ± 0.41e

4.00 ± 0.41e

5.00 ± 0.48B

Mean

7.17 ± 0.15A

4.69 ± 0.20B

4.56 ± 0.22B

 

Means sharing similar letters in a row or in a column are statistically non-significant (P > 0.05). Small letters represent comparison among interaction means and capital letters are used for overall means

Table 2

Mean values (±SE) of semen volume (ml) collected from bulls of control and treatment groups on different weeks

Weeks of Treatment

Group

Overall Mean

A (Control)

B (Treatment 1)

C (Treatment 2)

0

5.25 ± 0.95d-h

4.50 ± 0.29fgh

3.75 ± 0.48 h

4.50 ± 0.38D

5

3.38 ± 0.24 h

5.00 ± 0.41d-h

4.50 ± 0.29fgh

4.29 ± 0.26D

6

3.75 ± 0.48 h

5.75 ± 0.14c-g

4.75 ± 0.25e-h

4.75 ± 0.30CD

7

4.00 ± 0.41gh

6.50 ± 0.20a-e

6.00 ± 0.41b-f

5.50 ± 0.37BC

8

3.63 ± 0.38 h

6.50 ± 0.20a-e

6.75 ± 0.32a-d

5.63 ± 0.46ABC

9

3.63 ± 0.38 h

7.38 ± 0.24abc

7.38 ± 0.24abc

6.13 ± 0.55AB

10

3.75 ± 0.48 h

7.63 ± 0.24abc

7.88 ± 0.13ab

6.42 ± 0.59AB

11

3.63 ± 0.38 h

7.88 ± 0.31ab

8.00 ± 0.20a

6.50 ± 0.63A

12

3.63 ± 0.24 h

7.88 ± 0.13ab

8.13 ± 0.13a

6.54 ± 0.63A

Mean

3.85 ± 0.17B

6.56 ± 0.21A

6.35 ± 0.28A

 

Means sharing similar letters in a row or in a column are statistically non-significant (P > 0.05). Small letters represent comparison among interaction means and capital letters are used for overall means

Table 3

Mean values (±SE) of Mass Activity collected from bulls of control and treatment groups on different weeks

Weeks of Treatment

Group

Overall Mean

A (Control)

B (Treatment 1)

B (Treatment 2)

0

1.75 ± 0.25ef

1.50 ± 0.29f

1.50 ± 0.29f

1.58 ± 0.15E

5

1.75 ± 0.25ef

2.25 ± 0.25def

2.00 ± 0.00def

2.00 ± 0.12DE

6

2.00 ± 0.00def

2.25 ± 0.25def

2.00 ± 0.00def

2.08 ± 0.08CDE

7

2.00 ± 0.00def

2.50 ± 0.29c-f

2.50 ± 0.29c-f

2.33 ± 0.14CD

8

1.75 ± 0.25ef

2.50 ± 0.29c-f

2.75 ± 0.25b-e

2.33 ± 0.19CD

9

1.75 ± 0.25ef

3.00 ± 0.00a-d

3.00 ± 0.00a-d

2.58 ± 0.19BC

10

2.00 ± 0.00def

3.50 ± 0.29abc

3.50 ± 0.29abc

3.00 ± 0.25AB

11

2.00 ± 0.00def

3.75 ± 0.25ab

4.00 ± 0.00a

3.25 ± 0.28A

12

2.00 ± 0.00def

4.00 ± 0.00a

4.00 ± 0.00a

3.33 ± 0.28A

Mean

1.89 ± 0.05B

2.81 ± 0.15A

2.81 ± 0.15A

 

Means sharing similar letters in a row or in a column are statistically non-significant (P > 0.05). Small letters represent comparison among interaction means and capital letters are used for overall means

Analysis of variance indicated that motility percentage was highly significant (P˂0.01) in treated groups as compared to control. However, difference between group B and C was non-significant. Interaction between weeks and groups was also significant (Table 4). Analysis of variance reveled that sperm concentration is highly significant (P < 0.01) in both treated group as compared to control group (Table 5). However, difference between group B and C was non-significant. Interaction between weeks and groups was also significant. Mean value of live sperm percentage in control group A was lower as compared to b and C. Analysis of variance showed that there was significant effects on treated group as compared to control (Table 6). Overall mean value for HOST in control group A was lower as compared to treated groups (Table 7). Weekly analysis of variance showed that there was highly significant (P < 0.01) difference between treated and control group. Interaction between weeks and groups was also significant.
Table 4

Mean values (±SE) of Motility percentage collected from bulls of control and treatment groups on different weeks

Weeks of Treatments

Group

Overall Mean

A (Control)

B (Treatment 1)

C (Treatment 2)

0

68.75 ± 1.25c-f

65.00 ± 2.04ef

65.00 ± 0.00ef

66.25 ± 0.90C

5

63.75 ± 1.25f

70.00 ± 2.04b-f

66.25 ± 2.39def

66.67 ± 1.28C

6

67.50 ± 1.44def

68.75 ± 1.25c-f

67.50 ± 1.44def

67.92 ± 0.74BC

7

67.50 ± 1.44def

70.00 ± 2.04b-f

68.75 ± 1.25c-f

68.75 ± 0.90BC

8

66.25 ± 2.39def

73.75 ± 1.25a-e

75.00 ± 2.04a-d

71.67 ± 1.55AB

9

66.25 ± 2.39def

77.50 ± 1.44abc

77.50 ± 1.44abc

73.75 ± 1.86A

10

66.25 ± 2.39def

77.50 ± 1.44abc

77.50 ± 1.44abc

73.75 ± 1.86A

11

63.75 ± 2.39f

78.75 ± 1.25ab

80.00 ± 2.04a

74.17 ± 2.45A

12

61.25 ± 1.25f

81.25 ± 1.25a

82.50 ± 1.44a

75.00 ± 3.02A

Mean

65.69 ± 0.67B

73.61 ± 0.99A

73.33 ± 1.14A

 

Means sharing similar letters in a row or in a column are statistically non-significant (P > 0.05). Small letters represent comparison among interaction means and capital letters are used for overall means

Table 5

Mean values (±SE) of Sperm Concentration (Million/ml) of semen collected from bulls of control and treatment groups on different weeks

Weeks of Treatments

Group

Overall Mean

A (Control)

B (Treatment 1)

C (Treatment 2)

0

1626.8 ± 38.0d-g

1001.9 ± 182.9 k

1246.0 ± 108.5 h-k

1291.5 ± 101.3 F

5

1266.0 ± 93.8 g-k

1429.8 ± 29.1f-j

1354.0 ± 24.9f-k

1349.9 ± 036.6EF

6

1094.8 ± 102.2jk

1591.5 ± 16.0e-h

1472.5 ± 13.8e-i

1386.3 ± 071.2EF

7

1176.0 ± 41.6ijk

1696.8 ± 26.1def

1644.0 ± 39.4def

1505.6 ± 073.1DE

8

1171.3 ± 92.8ijk

1844.5 ± 83.8cde

1811.0 ± 38.3cde

1608.9 ± 101.4CD

9

1171.3 ± 92.8ijk

2071.3 ± 56.2abc

1988.5 ± 39.2bcd

1743.7 ± 127.3BC

10

1270.0 ± 72.5 g-k

2356.0 ± 47.7ab

2105.8 ± 45.8abc

1910.6 ± 143.1AB

11

1243.3 ± 52.8 h-k

2371.8 ± 46.2a

2299.0 ± 59.4ab

1971.3 ± 157.9A

12

1196.3 ± 46.5ijk

2405.0 ± 47.4a

2378.0 ± 58.4a

1993.1 ± 172.0A

Mean

1246.2 ± 32.7B

1863.2 ± 80.2A

1811.0 ± 67.3A

 

Means sharing similar letter in a row or in a column are statistically non-significant (P > 0.05). Small letters represent comparison among interaction means and capital letters are used for overall mean

Table 6

Mean values (±SE) of live percentage of spermatozoa collected from bulls of control and treatment groups on different weeks

Week of Treatments

Group

Overall Mean

A (Control)

B (Treatment 1)

C (Treatment 2)

0

83.75 ± 1.25a-d

77.50 ± 1.44 cd

76.25 ± 1.25 cd

79.17 ± 1.20BC

5

73.75 ± 2.39d

77.50 ± 1.44 cd

75.00 ± 2.04 cd

75.42 ± 1.14C

6

76.25 ± 2.39 cd

77.50 ± 1.44 cd

75.00 ± 2.04 cd

76.25 ± 1.09BC

7

73.75 ± 2.39d

77.50 ± 1.44 cd

76.25 ± 2.39 cd

75.83 ± 1.20C

8

76.25 ± 2.39 cd

80.00 ± 2.04bcd

82.50 ± 2.50a-d

79.58 ± 1.44BC

9

76.25 ± 2.39 cd

83.75 ± 2.39a-d

85.00 ± 2.89a-d

81.67 ± 1.78AB

10

73.75 ± 2.39d

86.25 ± 2.39abc

85.00 ± 2.89a-d

81.67 ± 2.16AB

11

73.75 ± 2.39d

90.00 ± 2.04ab

92.50 ± 1.44a

85.42 ± 2.71A

12

73.75 ± 2.39d

93.75 ± 1.25a

93.75 ± 1.25a

87.08 ± 2.98A

Mean

75.69 ± 0.85B

82.64 ± 1.12A

82.36 ± 1.33A

 

Means sharing similar letters in a row or in a column are statistically non-significant (P > 0.05). Small letters represent comparison among interaction means and capital letters are used for overall mean

Table 7

Mean values (±SE) of sperm membrane functional integrity (%) of control and treatment groups on different weeks

Week of Treatments

Group

Overall Mean

A (Control)

B (Treatment 1)

C (Treatment 2)

0

52.50 ± 1.44hi

51.25 ± 1.25i

53.75 ± 2.39ghi

52.50 ± 0.97 F

5

51.25 ± 1.25i

55.00 ± 2.04ghi

53.75 ± 2.39ghi

53.33 ± 1.12 F

6

51.25 ± 1.25i

62.50 ± 1.44e-h

57.50 ± 3.23f-i

57.08 ± 1.79EF

7

52.50 ± 1.44hi

66.25 ± 1.25c-f

63.75 ± 2.39d-g

60.83 ± 2.03DE

8

52.50 ± 3.23hi

68.75 ± 1.25a-e

67.50 ± 1.44b-f

62.92 ± 2.50CD

9

52.50 ± 3.23hi

72.50 ± 1.44a-e

71.25 ± 2.39a-e

65.42 ± 3.04BCD

10

55.00 ± 4.08ghi

73.75 ± 1.25a-d

71.25 ± 1.25a-e

66.67 ± 2.84ABC

11

55.00 ± 2.04ghi

77.50 ± 1.44ab

76.25 ± 1.25abc

69.58 ± 3.23AB

12

55.00 ± 2.04ghi

78.75 ± 1.25a

78.75 ± 1.25a

70.83 ± 3.47A

Mean

53.06 ± 0.75B

67.36 ± 1.59A

65.97 ± 1.62A

 

Means sharing similar letters in a row or in a column are statistically non-significant (P > 0.05). Small letters represent comparison among interaction means and capital letters are used for overall means

Discussion

This study was planned to examine the effect of supplementation of feed with flaxseed oil on libido and quality parameters of Nilli- Ravi buffalo bull’s semen. Poly unsaturated fatty acids are mainly present in mammalian sperm which are not synthesized in body so they should be supplemented [31].

In the present study, libido in treated bulls was increased from 9 to 12 weeks of treatment. These values are in line with Estienne et al., [11] reported in boars. This increment in the libido may be due to production of higher amount of testosterone hormone. As testosterone is a steroid hormone and synthesized by cholesterol. Flaxseed oil having polyunsaturated fatty acids such as olic acids and linolinic acids which are major source of cholesterol [22]. In ram testosterone level increases in short day, but by supplementation of flaxseed oil same concentration is obtained in summer months (Baiomy and Mottelib, 20e409).

The volume of semen volume was increased in from 9 to 12 months of treatment as compared to control animals. Similarly, increase in semen volume was observed in goats [9], rams [5] and quails [4], Rooke et al., [26]. In contrary to present study Gliozzi et al., [14], Adeel et al., [1], Gholami et al., [13] reported non-significant increase in semen volume in chicken, rabbit, buffalo bulls and Holstein bulls respectively. Flaxseed contains high value of omega-3 and omega-6 fatty acids which acts as antioxidant.

In present study overall mean values of mass activity were higher due to flax seed. Jafaroghli et al., [18] reported that supplementation with omega-3 fatty acids lead to significantly increase in mass activity of human semen. Improvement in mass activity of semen may be due to higher concentration and viablity sperm.

The importance of sperm motility has achieved a central role in the routine diagnosis of male fertility [3]. Motility percentage was significantly higher (p˂0.01) in treatment groups as compared to control group during 8–12 weeks of trial. Similar kind of results are reported in human [8], pigs [26], rams [5], goat [9, 28], rats [32] and boars [20]. Contrary to this study Adeel et al., [1] reported no improvement in motility of semen of buffalo bulls. Increased in motility percentage of semen may be due to fatty acids composition of sperm tail as reported by Mourvaki et al., [21] that flaxseed seed supplementation increased poly unsaturated fatty acids in rabbit sperm. This study showed that tail is major portion which is affected by supplementation of flaxseed supplementation. It can be assumed that poly unsaturated fatty acids present in flaxseed oil are involved in flagellar movement of sperm which in turn increases the motility percentage. The overall mean values of sperm concentration of treated groups were higher as compared to control. These results were similar in Stallion [29], goat [9], boars [11, 12], rams (Baiomy and Mottelib [5]) and men [12]. This increment in the sperm concentration in present study may be due to rapid spermatogenesis and antiapoptosis properties of supplementation of omega -3 fatty acids in feed. Supplementation of poly unsaturated fatty acids has positive effect on biosynthesis of prostaglandin and steroidogenesis. Poly unsaturated fatty acids have ability to affect the hypothalamic—pituitary axis and hormonal control of spermatogenesis.

The viability of sperm is considered as the useful parameters to differentiate between fertility and infertility conditions of males [15]. In the present study, lives sperm percentage was increased from 8 to 12 weeks of treatment. These findings are in close harmony as described by Rooke et al., [26], Dolatpanah et al., [9], Baiomy and Mottelib, [5], Mourvaki et al., [21],Gholami et al., [13], Al-Daraji et al., [4] and Esmaeili et al., [10]. The more number of live spermatozoa in ejaculates may be due to inhibition of apoptosis by action of poly unsaturated fatty acids as seen in culture off retinal photoreceptor and neuron [23].

The intact functional membrane of sperm is considered as the prerequisite for the sperm fertilizing ability due to successful acrosome reaction and binding to egg surface [7]. The functional integrity of sperm increased from 9 to 12 weeks of treatment in treated animals as compared to control. Similar to this study, comparable results were present in rabbit buck [21], buffalo bull [1], and in Holstein bulls Gholami et al., [13]. Increase in HOST positive spermatozoa was may be due to effect of poly unsaturated fatty acid on integrity of membrane [1].

Conclusion

It could be concluded that Supplementation of flaxseed oil has beneficial effect on semen quality parameter and libido of Nilli-Ravi buffalo bulls. This study further need to evaluate the effects of flax seed on cryopreservation and bull fertility.

Declarations

Acknowledgements

We are highly thankful to staff of semen production unit for their cooperation.

Authors’ contributions

First three authors played an active role in designing and conducting this trial. Similarly, parameter analysis, data analysis and manuscript preparation was performed by last two authors. All authors read and approved the final manuscript.

Competing interests

The authors have no conflict for the publication of this manuscript.

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors’ Affiliations

(1)
Department of Theriogenology, Faculty of Veterinary Sciences, University of Agriculture Faisalabad

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