THE INFLUENCE OF SELECTED FACTORS ON THE BEEF BULLS SPERM QUALITY USED FOR ARTIFICIAL INSEMINATION

Małgorzata Kunowska-Slósarz, Tomasz Przysucha, Marcin Gołębiewski, Jan Slósarz, Marek Balcerak
Department of Animal Breeding, Faculty of Animal Science, University of Life Sciences - SGGW, Poland

ABSTRACT

The aim of study was to examine the effect of bulls breed and collection month on characteristics of ejaculates: volume, concentration of spermatozoa, wave motion, percentage of live sperm in the ejaculate and percent of morphological defects of spermatozoa. The investigation was carried out on the 3454 beef bulls ejaculates collected from the following breeds: Limousine, Simmental, Piemontese, Charolaise, Blonde d’Aquitaine. Results reviled that breed and collection month significantly influenced volume, concentration of spermatozoa, wave motion, percentage of live sperm in the ejaculate and percent of morphological defects of spermatozoa obtained from beef bulls used for insemination. The highest volume of semen had Simmental bulls (5,69 ± 0,13 ml), the smallest volume of ejaculate characterized by semen collected from Piemontese bulls (3,05 ± 0,07 ml). Blonde d’Aquitaine breed had the highest concentration of spermatozoa (1545,57 ± 19,8 thous./ml). The Piemontese semen characterised by the smallest concentration (895,56 ± 15,45 thous./ml). Ejaculates collected in August and September had the largest volume, in January the lowest value of this trait was recorded. The highest concentration of spermatozoa had ejaculates collected in January. The lowest content of spermatozoa was recorded in August and September. Seasonal variability of sperm parameters was probably the result of a complex range of factors that can affect the animal organism, and the crucial is the air temperature.

Key words: beef bull, ejaculate, breed, artificial insemination, sperm quality.

INTRODUCTION

Artificial insemination is now one of the main biotechniques applied to cattle breeding, which accelerates genetic progress by insemination of cows with semen from the best bulls. Artificial insemination is also a method that allows the use of large-scale crossbreeding, where dairy cows are inseminated with semen from beef bulls in order to improve fattening and slaughter traits of the offspring [7]. Male fertility in Reproductive Centers, and above all the quality and quantity of sperm obtained from them, is a very important factor that may affect the effects of insemination. Numerous studies conducted in Poland and around the world are focused on the identification of factors that may interfere with the normal production of semen. The main factors are: environmental factors, physiological and genetic [1, 10, 14, 15]. Most of the researches conducted in Poland are based on evaluation of the impact of these factors on traits of dairy bulls semen, and study on the quality of beef bulls sperm practically has not been investigated yet.

The aim of presented study was to analyze the influence of selected factors (breed and month of collection) on the parameters of beef bulls semen used for insemination.

MATERIALS AND METHODS

The investigations were carried out on the 3454 beef bulls ejaculates collected from the following breeds (1210 ejaculates from Limousine (LM), 623 – Simmental (SM), 682 – Piemontese (PI), 541 – Charolaise (CH), 398 – Blonde d'Aquitaine (BD). All bulls were kept under the minimal standards of farm animal welfare for cattle studs. Animals were fed according INRA system with total mix ration depending on their breed and body weight. Ejaculates were obtained in 2004–2008 in Mazovian Centre of Animal Breeding and Reproduction in Łowicz. Frequency of semen collection mainly depended on the demands of the Centre for bull sperm. Ejaculates were collected by artificial vagina use. The following semen parameters were evaluated: volume of ejaculate (ml) (read from a calibrated semen reservoir), concentration of spermatozoa in the ejaculate (thous. / ml) – photometric method, sperm mass movement (in degrees: "+ + +" – strong waves (3), "+ +" – medium-wave (2), "+" – weak waves (1), "+" – barely marked fluctuation (-1), "–" – no wave (0), percentage of live sperm showing progressive motion, and sperm morphology (%) was carried out with Bydgoszcz method [3].

Statistical analysis of the data was performed using SPSS 12.0. (analysis of variance of the least squares method). The influence of the breed and the month of collection on sperm parameters were investigated. The significance of differences was estimated by Fischer test.

RESULTS

The data contained in Table 1 shows the diversity of semen parameters collected from analyzed breeds. The mean volume of ejaculates obtained from beef bulls was 4.31 ± 0.04 ml. In presented study, the highest volume of semen had Simmental bulls (mean 5.69 ± 0.13 ml), which was significantly different (P ≤ 0,01) than the average volume of the ejaculates of the other breeds. The smallest volume of ejaculate characterized by semen collected from Piemontese bulls, were average volume was 3.05 ± 0.07 ml, and statistically differed (P ≤ 0,01) from the average volume of ejaculates of the other examined breeds. All Piemnontese bulls in the study had muscle hypertrophy mutation. Similarly to Piemontese cattle, low volume of semen in our study had Blonde d'Aquitaine bulls (3.69 ± 0.09 ml).

Table 1. Ejaculate characteristics depending on the breed

Breed	N	Volume [ml]		Concentration [thous. / ml]		Sperm mass movement		% of sperm motility		Morphology [%]
		LSM	SE	LSM	SE	LSM	SE	LSM	SE	LSM	SE
		4.31	0.04	1274.36	8.99	2.33	0.02	75.44	0.25	1.68	0.06
BD	398	3.69A	0.09	1545.57A	19.80	2.53A	0.04	77.12A	0.55	1.68Aa	0.14
CH	541	4.90A	0.07	993.09AB	16.54	1.86A	0.03	70.07AB	0.46	1.55B	0.12
LM	1210	4.23A	0.05	1519.10BC	11.99	2.34A	0.02	72.96AC	0.33	2.40ABCD	0.08
PI	682	3.05A	0.07	895.56AC	15.45	2.18A	0.03	76.82BC	0.43	1.32aC	0.11
SM	623	5.69A	0.13	1418.50AC	30.33	2.76A	0.06	80.24AC	0.84	1.47D	0.21
LSM – least square mean SE – standard error A, B, C, D – are diffrent at P ≤ 0.01, a, b, c, d – are diffrent at P≤0.05

The results reviled that the average concentration of spermatozoa of all tested breeds was 1274.36 thous. / ml. The Piemontese semen characterized by the smallest concentration (mean 895.56 ±  15.45 thous. / ml) which was significantly different (P ≤ 0,01) from the spermatozoa concentration in ejaculates of other studied breed. Blonde d'Aquitaine breed had the highest concentration of spermatozoa (average 1545.57 ± 19.80 thous. / ml), which differed significantly (P ≤ 0.01) from the sperm concentration in ejaculates of Charolais, Simmental and Piemontese bulls.

In the Simmental bulls sperm mass movement was the strongest (average 2.76 ± 0.06) and differed statistically (P ≤ 0.01) from the other breeds. The weakest value of this trait was observed in the Charolaise breed (1.86 ± 0.03).

The average level of sperm motility was 75.44% ± 0.25 (Tab. 1). Similarly, to previous trait, also the highest percentage of sperm moving with advancing motion was found in Simmental breed (mean 80.24 ± 0.84%, P ≤ 0.01). Our study reviled that, the smallest value of this trait was recorded in Charolaise breed (mean 70.07% ± 0.46).

Also sperm morphology was differed between analyzed breeds. The percentage of sperm defects averaged 1.68 ± 0.06. Most defects of spermatozoa were registered in Limousine ejaculates (2.40% ± 0.08). The smallest percentage of morphological defects were observed in the semen of Piemontese breed (1.32% ± 0.11).

Table 2 summarized data concerning the characteristics of bulls semen in each month of the year. Ejaculates collected in August (average 4.58 ± 0.11 ml) and September (average 4.64 ± 0.11 ml) had the largest volume (Tab. 2). In the coming months, the semen volume was gradually decreasing until January, when the lowest value of this trait was recorded (mean 3.9 ml). From January gradual increase in the volume of obtained ejaculates was observed.

Table 2. Ejaculate characteristics depending on the month of collection

Month of collection	N	Volume [ml]		Concentration [thous. / ml]		Sperm mass movement		% of sperm motility		Morphology [%]
		LSM	SE	LSM	SE	LSM	SE	LSM	SE	LSM	SE
		4.31	0.04	1274.36	8.99	2.33	0.02	75.44	0.25	1.68	0.06
I	300	3.90	0.11	1382.51	24.70	2.36	0.04	76.62	0.70	1.90	0.17
II	254	4.14	0.11	1369.37	25.86	2.41	0.05	76.43	0.72	1.76	0.18
III	294	4.33	0.11	1360.33	24.48	2.53	0.05	77.41	0.68	1.69	0.17
IV	281	4.19	0.11	1314.38	25.36	2.40	0.05	76.76	0.71	1.87	0.18
V	301	4.25	0.10	1280.58	23.31	2.39	0.05	75.86	0.65	1.64	0.16
VI	272	4.42	0.11	1253.59	24.67	2.24	0.05	74.52	0.69	1.60	0.17
VII	303	4.44	0.11	1227.21	23.95	2.29	0.05	72.45	0.67	1.40	0.17
VIII	306	4.58	0.11	1115.56	23.87	2.26	0.05	74.08	0.66	1.45	0.17
IX	264	4.64	0.11	1153.24	25.77	2.26	0.05	76.02	0.72	1.59	0.18
X	302	4.48	0.11	1238.60	24.22	2.26	0.05	76.51	0.67	1.54	0.17
XI	260	4.11	0.12	1263.36	26.60	2.30	0.05	73.93	0.74	1.82	0.19
XII	263	4.23	0.12	1333.61	26.90	2.32	0.05	74.74	0.75	1.94	0.19
Significance of differences		P ≤ 0.01		P ≤ 0.01		P ≤ 0.01		P ≤ 0.01		NS
LSM – least square mean SE – standard error

The researches reviled that the highest concentration of spermatozoa had ejaculates collected in December (1333.61 ± 26.9 thous./ ml), January (1382.51 ± 24.7 thous. / ml), February (1369.37 ± 25.86 thous. / ml), March (1360.33 ± 24.48 thous. / ml) and April (1314.38 ± 25.36 thous. / ml) (Tab. 2).

The lowest content of spermatozoa in the beef bulls semen was recorded in August (1115.56 ± 23.87 thous. / ml) and September (1153.24 ± 25.77 thous. / ml), and the reduction of the semen concentration was also evident in May (Tab. 2). Our study confirms an inverse relation between the volume of semen and its concentration of spermatozoa in ejaculates.

The highest wave motion showed semen collected from January to May and in November and December (Tab. 2).

Also, the highest percentage of live sperm was found in the months from January to April and in September and October (Tab. 2). The lowest values of this trait coincided with the period from May to September and November to December. July was characterized by the lowest percentage of live sperm (mean 72.45%), and statistically (P ≤ 0.01) differed from the other months.

The number of reported changes in sperm morphology was also changed during the year, but were not statistically significant. The most anomalies in the construction of sperm was found in the semen in autumn and winter, while at least in the spring and summer (Tab. 2).

DISCUSSION

The male genotype is one of the traits determining the characteristics of bull’s semen [9, 17].

Fuerst-Waltl et al [6], who analyzed the semen of Simmental bulls recorded an annual average volume ejaculate of 5.8 ml in one breeding center and 6.9 ml in the second, which were slightly higher values than those obtained in the study.

Research showed, that the double muscling was associated with reduced fertility. Arthur [2] stated that the volume of semen collected from hypertrophic bulls was indeed smaller than the "normal" bulls, but the total number of spermatozoa in the ejaculates was not deviate from the norm. Also, Pileckas et al. [18], who observed changes in sperm parameters in different breeds: Holstein, Limousine, Salers x Aberdeen Angus, Simmental and Charolaise obtained comparable results. Much weaker sperm motility was evaluated in the study by Fuerst-Waltl et al [6].

Season is the term involving many factors such as temperature, humidity, length of light day, the quality of feed or welfare. The world's population of cattle is kept in different climate zones, so the impact of environmental factors on the animal organism was studied in different breeds and different countries. Many studies confirmed that, the season affected sperm production [13, 14, 19, 21], while in others there was no such an effect [4]. These differences are mainly due to the temperature distribution during the year, as well as the degree of adaptation to the climatic conditions of breed [22].

In the study of Janyk [11], where the dairy bulls were examined, the largest volume of semen was observed in the summer months: July, August and September, while the volume of ejaculates decreased in autumn and winter, which fits the results obtained in the present study. Similar observations were indicated by Drzażdżyński and Grodzki, [5] and Klupczyński et al. [13] who reported that the volume of the bulls semen during the summer months was much higher than during the winter months. Mroczek [16] presented the study on Simmental breed which also confirmed these observations.

The concentration of spermatozoa in ejaculates is a very important feature, because on the basis of this trait number of semen doses from ejaculate is estimated. Study of Miciński et al., [15] and Gumowski [8], confirmed that the highest sperm concentration was reported in the months from January to March. However, in the study of Mroczek [16], Simmental bulls had the highest concentration of spermatozoa in the spring months.

Janyk [11] and Gumowski [8] in their study observed a similar trend and showed reduced concentration of sperm in summer and autumn. The outside conditions directly affects the bulls testicles temperature and the whole process of spermatogenesis. Nevertheless, the influence of microclimatic factors on semen quality must be explain with caution as the sperm have been maturing in the male sexual ducts from 4 to 6 weeks, the features of ejaculates reflect environmental conditions about two months prior to collection. Considering our research, probably high temperatures in the summer had a negative effect on the sperm production and thus lowered the concentration of spermatozoa in ejaculates collected in August and September. There is no clearly defined the range of temperature, which would not disturb semen production. According to Jaśkowski [12] the optimum temperatures for spermatogenesis ranged between 10–20°C, Fuerst-Waltl et al. [6] indicated that 5–15°C and, Taylor et al., [21] find the optimum temperature of 15–20°C. Igna et al. [10] reported adverse effects of temperature (above 20°C) on basic parameters of Simmental bulls semen.

Our study reviled that the largest volume semen were characterized by ejaculates collected in the summer and early autumn period, while during this period the lowest concentration of sperm was observed, which confirms the results obtained by Gumowski [8].

The study reviled similar semen wave motion to results  reported by Janyk [11] who observed that semen of bulls had the highest wave motion from January to July. However, in studies of Mroczek [16] the strongest spermatozoa wave movement characterized the semen collected during the winter months, and the weakest in the summer months.

Stenzel and Kamieniecki [20], stated the increased percentage of sperm with normal movement in the winter and spring months. Jaśkowski [12] stated that this is probably a result of high temperatures during the summer. However, Gumowski [8] in their study identified no seasonal changes in the number of live sperm in the ejaculates during the year.

CONCLUSIONS

To summarized breed, and months of semen collection had an effect on ejaculate volume, sperm concentration, the mass movement of sperms, percentage of live sperm and their morphology. Seasonal variability of sperm parameters was probably the result of a complex range of factors that can affect the animal organisms, and the crucial is the air temperature.

REFERENCES

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Accepted for print: 26.03.2015

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Małgorzata Kunowska-Slósarz
Department of Animal Breeding, Faculty of Animal Science, University of Life Sciences - SGGW, Poland
ul. Ciszewskiego 8
02-786 Warsaw
Poland
email: malgorzata_kunowska_slosarz@sggw.pl

Tomasz Przysucha
Department of Animal Breeding, Faculty of Animal Science, University of Life Sciences - SGGW, Poland
Ciszewskiego 8, 02-786 Warszawa, Poland
Phone/fax: 48 22 5936536
email: tomasz_przysucha@sggw.pl

Marcin Gołębiewski
Department of Animal Breeding, Faculty of Animal Science, University of Life Sciences - SGGW, Poland
ul. Ciszewskiego 8
02-786 Warsaw
Poland

Jan Slósarz
Department of Animal Breeding, Faculty of Animal Science, University of Life Sciences - SGGW, Poland
ul. Ciszewskiego 8
02-786 Warsaw
Poland

Marek Balcerak
Department of Animal Breeding, Faculty of Animal Science, University of Life Sciences - SGGW, Poland
ul. Ciszewskiego 8
02-786 Warsaw
Poland

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