Semen Quality of Bulls as Influenced by Breed, Body Condition Score and Ascorbic Acid under Heat Stress

Muamber Shah1, Muhammad Subhan Qureshi1, Rifat Ullah Khan1, Muhammad Mobashar1, Mubashar Ali Khalique2, Irfan Khattak3, Ambrina Tariq4, Ijaz Ahmad1 and Shabana Naz5,*

1Faculty of Animal Husbandry and Veterinary Sciences, The University of Agriculture, Peshawar

2Faculty of Veterinary and Animal Sciences, University of Poonch, Rawlakot, Azad Kashmir

3College of Veterinary Science and Animal Husbandry, Abdul Wali Khan University, Mardan

4Civil Veterinary Hospital, Dera Ismail Khan, Directorate of Livestock and Dairy

Development, Peshawar

5Department of Zoology, Govt. College University, Faisalabad

ABSTRACT

This study evaluated the effect of breed, body condition score and ascorbic acid supplementation on the semen quality of breeding bulls. A total of 8 adult (≥2 year) bulls (2 each from Jersey, Friesian, Sahiwal and cross bred) were selected with body condition score (BSC) at a scale of (0-5). The results showed that mean semen volume was significantly (P<0.05) high in Friesian bull. Sperm concentration and motility were significantly (P<0.05) high in Jersey bull while live sperm percentage was significantly (P<0.05) high in Jersey bulls and sperm abnormality was significantly (P<0.05) high in Sahiwal and Friesian-Sahiwal cross. In addition, semen quality was improved in all the breeds after supplementation of vitamin C. There was no significant effect of BCS on the sperm quality of bulls. The results give important information regarding the semen quality of different breeds of bulls under heat stress condition.

Article Information

Received 01 November 2018

Revised 11 December 2018

Accepted 20 December 2018

Available online 18 June 2019

Authors’ Contribution

MS, MSQ and RUK conducted the study. MM, MAK and IK wrote and edited the manusript. IA conducted the statistical analysis. SN revised the manusript.

Key words

Bulls, Semen, Body condition score, Motility.

DOI: http://dx.doi.org/10.17582/journal.pjz/2019.51.5.1699.1703

* Corresponding author: drshabananaz@gcuf.edu.pk

0030-9923/2019/0005-1699 $ 9.00/0

Copyright 2019 Zoological Society of Pakistan

Introduction

When the animal is exposed to high environmental temperature a state of heat stress could occur which affects the quality of semen (Marai et al., 2010). Heat stress may directly affect reproductive performance in the form of impaired spermatogenesis and lower testosterone level (Murugaiyah, 1992). In Pakistan, during summer, the atmospheric temperature ranges from 30-45oC. For exotic breeds, high ambient temperature has adverse effect on the reproductive performance (Ihsanullah et al., 2017). Ascorbic acid works as a natural antioxidant and is found in elevated level in seminal fluid (Thiele et al., 1995). Vitamin C also plays an important role in reproduction and showed significant results (Luck et al., 1995; Khan et al., 2012). Its role in the physiological reproduction had been undefined. Ascorbic acid prevents sperm oxidative damage by keeping genetic integrity of sperm cells. Considering the merits and future potential and benefits of using ascorbic acid, its effects on the semen parameters are needed to be explored. Therefore, the present study was designed to investigate the effects of different levels of ascorbic acid and body condition score (BCS) on semen quality and semen traits of bulls. The present study assessed the effect of heat stress on the semen quality under different BCS and the ameliorative effect of ascorbic acid supplementation on semen traits of different breeds of bulls.

 

Materials and methods

A total of 8 adult (≥2 year) bulls (2 each from Jersey, Friesian, Sahiwal and cross bred) were selected. Body condition score was recorded at a scale of (0-5) points as described by Ihsanullah et al. (2016). Further 12 animals of different breeds were selected and divided into four treatment groups. Ascorbic acid was supplemented in feed at the rate of 0, 50, 100, and 200 mg/day of feed. The experiment was continued for 3 weeks.

Semen evaluation

Semen was collected from the experimental bulls early in the morning using artificial vagina (AV). Soon after collection, semen was transferred to water bath with a temperature of 35°C for further investigation. The collection was done twice a week i.e., Monday and Thursday. The semen samples were subjected to determination of semen volume, sperm concentration, percentage of motile spermatozoa, live dead and sperm morphology as described by Majid et al. (2015).

Temperature and humidity index (THI)

The THI was calculated using the formula of Ihsanullah et al. (2017) as follow:

THI = (0.8 × Tmix) + [(H/100) × (Tmix − 14.4)] + 46.4

Where, Tmix is maximum temperature in centigrade and H is percent relative humidity.

Different levels of THI in dairy breeds: THI=< 72 (no stress), THI = 72-79 (mild stress), THI = 80-89 (moderate to high stress) and THI = > 96 (very danger and death may occur).

Statistical analyses

Data was statistically analyzed with the help of statistical software, Statistix (version 8.1). Means of control and treated groups were compared by using one way analysis of variance (Steel et al., 1997).

 

Results

Among the different breeds, maximum semen volume was produced by Holstein Friesian (HF) bulls followed by Jersey, Friesian Sahiwal Crossed (FSC) and Sahiwal as given in Table I. The overall mean volume of semen was also significantly higher (P<0.05) in HF breed as compared to other breeds in this study even in condition of high temperature and humidity. The results also presented that the volume of the semen in all breeds was high during first week and then reduced with an increase in temperature and humidity index. The concentration of semen was also significantly different (P<0.05) in different breeds of bulls in this study. Significantly, the highest semen concentration was found in Jersey bulls followed by HF, FSC and Sahiwal even at in high temperature and humidity. The overall mean of high concentration was also significantly high in sperm of Jersey bulls as shown in Table I. The result presented in Table I describes sperm percent motility of different breeds of bulls in different weeks at high temperature and humidity. The sperm percent motility was significantly (P<0.05) increased in Jersey bulls followed by the HF, FSC and Sahiwal breeds of bulls. The overall mean of sperm percent motility was also significantly high in Jersey breed of bulls compare to others. Results on live percentage of sperm among different breeds during various weeks at high temperature and humidity are shown in Table II. Live percentage of sperm was significantly (P<0.05) higher in Jersey breed of bulls followed by the HF, FSC and Sahiwal breed. The overall live percentage was also significantly high in Jersey bulls. Sperm primary abnormality of different breeds of bulls at different weeks in different temperature and humidity are shown in Table II. However, FSC bull breed showed significantly (P<0.05) greater sperm primary and secondary abnormalities at high temperature and humidity as compared to other bull breeds.

 

Table I.- Mean values of semen traits of different bull breeds at different temperature and humidity index (n=64).

Breed	Parameters	Week 1	Week 2	Week 3	Week 4	Overall mean
Friesian	Volume (ml)	6.26±0.14a	6.20±0.13a	5.75±0.15a	5.50±0.28a	5.68±0.03a
	Concentration (106/ml)	1375.0±14.43b	1375.0±14.43b	1300.0±28.86b	1275.0±14.43b	1331.3±1.04b
	% Motility	89.00±0.28b	88.50±1.15b	88.50±0.28b	87.50±0.28b	88.40±0.05b
Jersey	Volume (ml)	5.35±0.14b	5.30±0.12b	5.25±0.10b	5.25±0.14a	5.50±0.00ab
	Concentration (106/ml)	1725.0±14.43a	1675.0±14.43a	1600.0±28.86a	1600.0±28.86a	1650.00±7.21a
	% Motility	92.00±0.28a	91.50±0.86a	91.50±0.57a	91.50±0.86a	91.63±0.20a
Sahiwal	Volume (ml)	5.22±0.14b	5.16±0.10b	5.00±0.09c	4.75±0.28b	5.06±0.10c
	Concentration (106/ml)	900.00±14.43d	875.00±14.43d	875.00±28.86d	825.00±14.43d	868.75±10.8d
	% Motility	85.50±0.57d	83.00±0.86b	83.00±1.15c	82.50±0.28d	83.50±0.28d
FSC**	Volume (ml)	5.50±0.14b	5.50±0.12ab	5.25±0.28bc	5.00±0.14a	5.38±0.14c
	Concentration (106/ml)	1200.0±28.86c	1200.0±57.73c	1150.0±28.86c	1125.0±14.43c	1168.8±18.1c
	% Motility	90.00±0.28c	86.50±1.15a	85.00±0.28c	84.50±0.57c	86.53±0.43c
THI		83.68	84.75	85.22	86.16	84.95

a,b,c,d, means with different superscripts within the same column for the same parameter differ significantly (P<0.05) for different breeds. THI, Temperature and humidity index; FSC, Fresian Shiwal cross; THI, temperature and humidity index.

 

The results present in Table II describes the sperm secondary abnormality of different breeds of bulls at different weeks at high temperature and humidity showing that sperm secondary abnormality was significantly (P<0.05) high in FSC breed of bulls followed by Sahiwal, Friesian and Jersey breeds. The overall sperm secondary abnormality was also significantly (P<0.05) high in cross breed bulls as compare to other breeds in this study.

The ascorbic acid supplementation significantly (P<0.05) increased the volume of semen and sperm concentration in bulls. The highest volume of semen and sperm concentration was produced by the dose rate of 200 mg as compared to non-supplemented or low supplemented group as shown in Table III. The overall mean volume was also significantly high in the same group.

The result of the sperm concentration of different group of bulls at different weeks is given in Table III. The sperm concentration was significantly high (P<0.05) in all four weeks in group (C200) bulls as compare to all other groups in the study. The overall mean of sperm concentration was also significantly high in group (C 200) of bulls. However, Ascorbic acid supplementation didn’t influence live percent motility of sperm among different bull breeds (Table IV).

 

Table II.- Mean values of live percent and abnormalities percentage of sperm of different bull breeds at different temperature and humidity index (n=64).

Breed	Parameters	Week 1	Week 2	Week 3	Week 4	Overall mean
Friesian	Live %	93.00±0.57a	91.50±0.28b	90.50±0.28b	90.50±0.28b	91.37±0.07b
	ABN % (P)	5.50±0.28c	6.00±0.57c	6.00±0.57b	7.00±0.57bc	6.12±0.36c
	ABN % (S)	13.50±0.28 c	13.50±0.86b	14.00±1.15a	15.00±0.57b	14.00±0.72b
Jersey	Live %	93.50±0.28b	92.50±0.28a	92.00±0.28a	90.50±0.57a	92.12±0.07a
	ABN % (P)	3.00±00d	3.50±0.28d	5.50±0.28d	5.50±0.28d	4.37±0.07d
	ABN % (S)	13.50±0.28c	13.50±0.57b	13.50±0.86a	13.00±0.28b	13.37±0.36b
Sahiwal	Live %	89.50±0.57c	89.50±0.28c	89.00±0.86b	89.00±0.57c	89.25±0.14d
	ABN % (P)	7.50±0.28b	7.50±00b	9.00±0.57a	10.00±0.28b	8.50±0.14b
	ABN % (S)	15.00±0.28b	15.50±0.57a	16.50±0.57a	17.00±0.86ab	16.00±0.14a
FSC	Live %	91.50±0.28b	90.50±0.75c	90.50±0.28b	89.00±0.28b	90.37±0.21bc
	ABN % (P)	8.50±0.28a	10.50±0.28a	10.50±0.57a	11.00±0.86a	10.12±0.21a
	ABN % (S)	15.50±0.28a	16.50±0.28a	17.50±0.28a	17.50±0.86a	16.75±0.28a
THI		83.68	84.75	85.22	86.16	84.95

a,b,c,d, means with different superscripts within the same column for the same parameter differ Significantly (P<0.05) for different breeds. ABN (P), primary abnormality; ABN (S), secondary abnormality; FSC, Fresian Sahiwal cross.

 

Table III.- Effect of different supplementations of ascorbic acid on mean values of semen traits of different bull breeds at different weeks (n=96).

Group	Parameters	Week 1	Week 2	Week 3	Week 4	Overall mean
C 00	Volume (ml)	4.66±0.16b	4.83±0.16b	4.83±0.16b	5.06±0.06b	4.83±0.04b
	Concentration (106/ml)	1266 .7±28.86b	1276.7±44.09b	1300.0±14.53b	1316.7±16.66b	1290.3±16.4b
	% Motility	89.00±0.57a	89.66±1.20a	88.00±0.57a	88.66±0.33a	88.83±0.33a
C 50	Volume (ml)	5.50±0.28a	5.50±0.28a	5.50±0.28a	5.50±0.28a	5.50±0.14a
	Concentration (106/ml)	1300.0±33.33ab	1300.0±28.86ab	1316.7±28.86b	1333.3±16.66b	1312.7±21.6b
	% Motility	86.00±0.57a	88.00±0.57a	88.33±0.88a	89.33±0.66a	87.91±0.58a
C 100	Volume (ml)	5.50±0.28ab	5.50±0.28b	5.33±0.33ab	5.66±0.33b	5.50±0.14ab
	Concentration (106/ml)	1300.0±28.86b	1300.3±16.66ab	1333.0±28.86b	1333.3±16.66b	1316.7±8.33b
	% Motility	87.66±2.02a	87.66±0.33a	88.66±0.88a	89.33±0.66a	88.33±0.79a
C 200	Volume (ml)	5.66±0.28a	6.00±0.66a	6.00±0.57a	6.36±0.44a	6.01±0.49a
	Concentration (106/ml)	1383.3±44.09a	1383.3±18.86a	1400.0±16.66a	1433.3±44.09a	1400.3±3.94a
	% Motility	89.00±0.66a	88.66±0.66a	88.33±0.57a	88.33±0.33a	88.58±0.44a

a,b, means with different superscripts within the same column for the same parameter differ significantly (P<0.05)for different supplementation level. C 00, C50, C100 and C200 denote different vitamin C supplementation at rate of 0, 50, 100 and 200 mg/day of feed.

 

Table IV.- Effect of different levels of ascorbic acid on sperm live parentage, sperm primary and secondary abnormalities of different bull breeds (n=96) during different weeks.

Group	Parameters	Week 1	Week 2	Week 3	Week 4	Overall mean
C 00	Live %	92.00±0.57	91.33±1.20	91.00±0.57a	90.66±0.33a	91.08±0.36a
	ABN % (P)	9.66±0.88	8.00±0.66	7.33±0.33a	6.66±0.57a	7.33±0.36a
	ABN % (S)	14.00±0.57	15.33±0.88	12.00±0.57a	11.33±0.88a	13.16±0.46a
C 50	Live %	91.33±0.88	93.33±1.33	94.00±1.15a	90.33±0.33a	91.41±0.36a
	ABN % (P)	8.66±0.88	7.00±0.57	8.33±0.66a	5.33±0.33b	7.20±0.16ab
	ABN % (S)	13.66±1.76	13.00±0.57	12.00±o.57a	11.33±0.66a	12.50±0.76a
C 100	Live %	93.00±0.57	90.33±0.33	92.00±1.15a	90.33±0.33a	91.16±0.30a
	ABN % (P)	8.33±0.33	6.33±0.33	8.33±0.33a	5.33±0.33b	7.18±0.22ab
	ABN % (S)	13.66±0.88	13.33±0.88	13.66±0.88a	12.00±0.57a	13.16±0.08a
C 200	Live %	93.00±1.00	91.00±0.57	92.00±0.57a	91.33±0.33a	91.75±0.14a
	ABN % (P)	9.33±0.88	7.66±0.33	6.33±0.33b	6.00±0.57b	7.16±0.22b
	ABN % (S)	14.00±1.15	14.00±0.57	12.00±0.57a	11.33±0.33a	12.83±o.16a

Values with different superscripts within the same column differ significantly (P<0.05) for different supplementation levels. C00, C50, C100 and C200 represent 0, 5, 100 and 200 mg/feed day. ABN (P), primary abnormality; ABN (S), secondary abnormality.

 

Table V.- Mean values of different semen parameters of different bull breeds under variable body condition score (n=64).

Breed	BCS	Volume (ml)	Concent. (106/ml)	% Motility	Live %	% ABN (P)	% ABN (S)**
Friesian	3.25	6.05±0.21	1350±20.41	87.75±1.10	91.00±0.40	7.00±0.40	15.25±0.47
	3.50	5.77±0.27	1325±32.27	88.25±1.31	92.00±0.81	6.50±0.28	15.50±0.64
Jersey	3.25	6.05±0.21	1687.5±23.93	89.25±1.43	91.25±0.62	4.25±0.25	12.00±0.40
	3.75	5.65±0.11	1650.0±35.35	88.75±1.03	89.00±0.40	4.25±0.62	12.00±0.70
Sahiwal	3.00	5.30±0.28	837.50±23.93	82.75±0.47	88.75±0.47	5.75±0.47	12.50±0.50
	3.25	5.00±0.14	837.50±12.50	84.50±0.50	87.50±0.28	6.50±0.86	13.50±0.00
FSC	3.00	5.60±0.07	1237.5±12.50	85.25±1.49	87.50±1.32	9.75±0.47	17.25±0.47
	3.5	5.57±0.13	1237.5±37.50	83.50±2.21	89.00±0.90	8.50±0.28	17.75±0.47

Mean values within the same column for the same parameters are not significantly different (P>0.05). BCS, body score condition; ABN (P), primary abnormality; ABN (S), secondary abnormality; FSC, Friesian Sahiwal cross.

 

The results revealed in Table IV that there was non-significant (P>0.05) difference in live percentage of sperm in different groups of bulls with different levels of ascorbic acid supplementation. Furthermore, primary abnormality in sperm among different bull breeds during various weeks was significantly higher (P<0.05) in control group while secondary abnormalities remained unaffected among treatments.

The different semen parameters like semen volume and concentration and percent motility and live percentage, primary abnormality and secondary abnormality of sperm of different bull breeds at various weeks were not significantly affected by variable body condition score (Table V).

 

Discussion

High environmental temperature has negative effects on fertility of bulls. In a previous study, it was reported that Holstein Friesian bulls are mostly affected by heat stress as compared to other breeds (Garacia et al., 2005). In the present study, sperm concentration was significantly higher in Jersey bulls compared to other breeds. This is in agreement with findings of Ahmad et al. (1993) and Shaha et al. (2008) who also found higher concentration of sperm in Jersey bulls. Similarly, in the present study, Jersey bulls also had lesser primary and secondary abnormalities in comparison with other bull breeds. While FSC bulls had significantly higher primary and secondary sperm abnormalities, which might be linked with its genetic disturbance and imbalance. It was also observed that live percentage and percent motility of sperm was significantly high in Jersey bull breed and high dead sperm ratio was found in Sahiwal breed. The high concentration of sperm, less dead percentage of sperm, high motility and lesser primary and secondary abnormality in sperm of Jersey breed showed that Jersey breed is lighter and very favorable for the subtropical environment. According to the breeding policy in Pakistan, the semen of Jersey breed is suitable for local cattle breeds.

In the current study, the semen volume was significantly increased and the sperm primary abnormalities were decreased in the ascorbic acid supplemented groups of bulls. Yousef et al. (2003) also observed similar pattern of semen volume with ascorbic acid treatment in rabbits. Similarly, higher sperm concentration was found in bulls at ascorbic acid level of C200. Dawson et al. (1987) also found significantly higher sperm concentration in human with higher level of ascorbic acid. Interestingly, sperm motility, live percentage of sperm and sperm abnormalities were not influenced by ascorbic acid treatment.

Animals with high BCS have highest sperm reserve count as well as sperm count (Salisbury et al., 1978). In the present study, all the semen parameters mentioned above at different weeks under variable BCS of different breeds’ bulls were not significantly different. Our results are in contrast with the results of Salisbary et al. (1978). The reason could be that we have collected data from semen processing unit, where all high quality semen producing animals were kept and the difference among the body condition score were not high. All the animals kept in SPU were healthy and there were no weak or obese animals which could not make any statistically difference in the semen traits.

 

Conclusion

Ascorbic acid should be supplemented to all bulls used for artificial insemination to improve the semen quantitative and qualitative characteristics under heat stress. The body condition score within range of (3-3.75) had no significance effect on semen traits. Jersey breed of cattle is the best adapted summer stressed breed.

 

Statement of conflict of interest

The authors declare no conflict of interest.

 

References

Dawson, E.B., Harris, W.A., Rankin, W.E., Charpentier, L.A. and McGanity, W.J., 1987. Effect of ascorbic acid on male fertility. Annls. N.Y. Acad. Sci., 498: 312-323. https://doi.org/10.1111/j.1749-6632.1987.tb23770.x

Ihsanullah, Qureshi, M.S., Suhail, S.M., Akhtar, S. and Khan, R.U., 2017. Postpartum ovarian activities, blood metabolites and milk yield are influenced by changing levels of thermal stress in crossbred dairy cows. Int. J. Biometerol., 61: 1561-1569. https://doi.org/10.1007/s00484-017-1335-z

Khan, R.U., Naz, S., Nikousefat, Z., Selvaggi, M., Laudadio, V. and Tufarelli, V., 2012. Effect of ascorbic acid in heat-stressed poultry. World’s Poult. Sci. J., 68: 477-490. https://doi.org/10.1017/S004393391200058X

Luck, M., Jeyaseelan, R.I. and Scholes, R.A., 1995. Ascorbic acid and fertility. Biol. Rep., 52: 262-266. https://doi.org/10.1095/biolreprod52.2.262

Marai, M. and Haeeb, A.A., 2010. Buffalo’s biological functions as affected by heat stress-A review. Livest. Sci., 127: 89-109. https://doi.org/10.1016/j.livsci.2009.08.001

Majid, A., Qureshi, M.S. and Khan, R.U., 2015. In vivo adverse effects of alpha-tocopherol on the semen quality of male bucks. J. Anim. Physiol. Anim. Nutr., 99: 841-846. https://doi.org/10.1111/jpn.12284

Murugaiyah, M., 1992. Changes in the semen characteristics of Kambing Katjan crossbreed buck under hot and humid environmental temperatures. In: Recent advances in goat production (ed. R.R. Lokeshan). Proc. 5th Int. Conf. Goats, ND, pp. 1126-1129.

Salisbury, G.W., Vandemark, N.L. and Lodge, J.R., 1978. Physiology of reproduction and artificial insemination of cattle. W.H. Freeman, Sanfrancisco, pp. 574.

Shaha, S.P., Alam, M.G., Khatun, M. and Ahmadi, U., 2008. Breeding and soundness of stud bulls. Bangladesh Vet., 25: 51-61. https://doi.org/10.3329/bvet.v25i2.4618

Steel, R.G.D., Torrie, J.H. and Dieky, A., 1997. Principles and procedures of statistics, 3rd ed. McGraw Hill Book Co. Inc., N.Y.

Thiele, J.J., Friesleben, H.J., Fuchs, J. and Ochsendorf, F.R., 1995. Ascorbic acid and urate in human seminal plasma: Determination and interrelationships with chemiluminescence in washed semen. Hum. Rep., 10: 110-115. https://doi.org/10.1093/humrep/10.1.110

Yousef, I., Abdullah, G.A. and Kamel, K.I., 2003. Effect of ascorbic acid and vitamin E supplementation semen quality and biochemical parameters of male rabbits. J. Anim. Reprod. Sci., 76: 99-111. https://doi.org/10.1016/S0378-4320(02)00226-9