alexa <p>Semen Characteristics of Normal and Low Grade Ejaculates in Ongole (Bos indicus) Bulls</p> | OMICS International
ISSN: 2157-7579
Journal of Veterinary Science & Technology

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Semen Characteristics of Normal and Low Grade Ejaculates in Ongole (Bos indicus) Bulls

Manda Srinivas1*, Makkena Sreenu2, Srilatha CH3, Babu Rao K4 and Naidu KS5

1Department of Veterinary Gynaecology and Obstetrics, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India

2Deparment of Veterinary Surgery and Radiology, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India

3Department of Veterinary Pathology, College of Veterinary Science, Tirupati, Andhra Pradesh, India

4Livestock Research Station, LAM, Guntur, Andhra Pradesh, India

5Sri Venkateswara Veterinary University, Tirupati, Andhra Pradesh, India

*Corresponding Author:
Manda Srinivas
Associate Professor
Department of Veterinary Gynaecology and Obstetrics
NTR College of Veterinary Science, Gannavaram
Andhra Pradesh-521 102, India
Tel: +919951332132
E-mail: [email protected]

Received Date: February 29, 2016; Accepted Date: October 07, 2016; Published Date: October 12, 2016

Citation: Srinivas M, Sreenu M, Srilatha CH, Rao BK, Naidu K (2016) Semen Characteristics of Normal and Low Grade Ejaculates in Ongole (Bos indicus) Bulls. J Vet Sci Technol 7:391. doi: 10.4172/2157-7579.1000391

Copyright: © 2016 Srinivas M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Abstract

The normal and low-grade ejaculates collected from the eight bulls under study were evaluated of various semen parameters. The overall means of HOS-G sperm was recorded as 56.50 ± 0.50 and 30.35 ± 0.66 percent for HOS positive and acrosome positive (HPAP), 17.30 ± 0.54 and 19.48 ± 0.66 percent for HOS positive and acrosome negative (HPAN), 18.73 ± 0.54 and 30.11 ± 0.89 percent for HOS negative and acrosome positive (HNAP) and 7.64 ± 0.47 and 19.63 ± 0.94 percent for HOS negative and acrosome negative (HNAN) in fresh semen of normal and low grade ejaculates, respectively. The overall mean sperm penetration distance was recorded as 19.75 ± 0.19 and 14.40 ± 0.19 mm/20 min in normal and low grade ejaculates (Table 1) with a highly significant difference between them (P<0.01). The sperm penetration distance (mm/20 min) differed only amongst bulls of low grade ejaculates and not among those of normal ejaculates. It was concluded from the present study that routine semen evaluation tests which are mainly subjective in nature suffered human bias and have limited ability to predict the fertilizing capacity of the sperm, hence still more sensitive tests like functional assessment of plasma membrane and acrosome membrane (HOS-G Test) and sperm penetration distance in synthetic medium are essential to predict optimum fertility.

Keywords

Semen characteristics; HOS-G; Sperm penetration distance; Normal ejaculates; Low-grade ejaculates; Ongole bulls

Introduction

Reproductive soundness evaluation facilitates the elimination of sub-fertile sires from breeding programmes. Andrological investigation of breeding bulls is important to evaluate them for their fertility which includes scrotal and testicular biometry, ultrasonography of male reproductive tract and testes and laboratory evaluation of semen quality with fertility [1]. It has been noticed that in indigenous as well as cross-bred bulls a huge percentage of ejaculates are discarded due to high proportion of abnormal sperms and their inability to withstand to freezing [2,3]. Earlier studies on semen from indigenous Gir and Jafrabadi bulls was observed to be consistently poor in its quality and freezability, in spite of proper inputs in health care, feeding and management. The quality of low-grade ejaculates could be improved by filtration methods, which removes the dead, non-motile and abnormal sperms. Thus studies on low grade ejaculates are of immense value in exploiting the breeding potential of genetically superior sires giving poor semen which was destined to be discarded [4]. Therefore, keeping in view of the above facts, the present study was attempted to evaluate the membrane integrity and sperm penetration distance of normal and low grade ejaculates in Ongole breeding bulls.

Materials and Methods

Semen was collected from eight bulls under study once in a week between 6.00 AM and 7.00 AM with the standard artificial vagina using an anoestrus cow as a dummy. Each time two ejaculates were collected at a gap of 20 to 30 minutes after allowing one or two false mounts. Soon after the collection, the semen collection tubes were numbered and transferred to a water bath at 37°C. During the period under report a total of 549 ejaculates were collected from the eight bulls under study they included normal ejaculates and low grade ejaculates discarded at various stages of semen processing. Ejaculates having initial motility between 40 and 50% were classified as low grade as per the method of Ahmed et al. [5]. Functional membrane integrity was assessed by hypo-osmotic swelling-Giemsa (HOS-G) test. 1 ml of hypo osmotic swelling solution (100 mOsmol/L) was mixed with 0.1 ml of semen and incubated at 37°C for 1 hour. Similarly, 0.1 ml of semen was incubated in isotonic solution (300 mOsmol/L). After incubation at 37°C for 30 minutes one drop of well-mixed sample was smeared on a warm glass slide and then stained with Giemsa. Minimum of 200 spermatozoa were observed in a phase-contrast microscope under oil immersion and expressed in percentage for different subpopulations. Sperm penetration distance was carried out as per the method described by Kumar [6]. Polyacrylamide gel a synthetic migration medium was prepared as per the method described by Lorton et al. [7] with slight modification.

Results

The overall means of HOS-G sperm was recorded as 56.50 ± 0.50 and 30.35 ± 0.66 percent for HOS positive and acrosome positive (HPAP), 17.30 ± 0.54 and 19.48 ± 0.66 percent for HOS positive and acrosome negative (HPAN), 18.73 ± 0.54 and 30.11 ± 0.89 percent for HOS negative and acrosome positive (HNAP) and 7.64 ± 0.47 and 19.63 ± 0.94 percent for HOS negative and acrosome negative (HNAN) in fresh semen of normal and low grade ejaculates, respectively (Table 1).

Parameter Ejaculate Quality 408 409 417 424 426 465 1011 1027 Over All Mean
HOS-G HPAP(%) Normal 59.90 ± 0.55a 56.70± 1.41ab 58.70 ± 1.12ab 55.60 ± 1.60b 57.60 ± 1.26ab 55.40 ± 1.30b 57.90 ± 0.72ab 50.20 ± 0.83c 56.50 ± 0.50a
Low Grade 35.00 ± 1.89ab 28.50 ± 0.81cd 31.10 ± 2.00bc 30.70 ± 1.43bc 30.70 ± 1.16bc 25.20 ± 1.37d 36.70 ± 1.45a 24.90 ± 1.17d 30.35 ± 0.66b
HOS-G HPAN(%) Normal 13.50 ± 0.43c 20.30 ± 1.57ab 16.40 ± 1.62bc 16.00 ± 0.92bc 15.90 ± 1.41bc 20.80 ± 1.37a 16.10 ± 1.03bc 19.40 ± 2.13ab 17.30 ± 0.54b
Low Grade 18.80 ± 1.55abc 14.70 ± 1.31c 17.90 ± 1.92bc 17.00 ± 1.92bc 21.10 ± 1.53ab 20.90 ± 1.39ab 21.50 ± 1.38ab 23.90 ± 2.48a 19.48 ± 0.66a
HOS-G HNAP(%) Normal 22.50 ± 0.78a 16.40 ± 1.19bc 20.30 ± 0.73ab 22.60 ± 1.19a 18.70 ± 1.52abc 14.70 ± 1.09c 17.70 ± 1.10bc 16.90 ± 2.34bc 18.73 ± 0.54b
Low Grade 32.60 ± 1.38b 42.90 ± 1.35a 28.20 ± 1.25bc 32.90 ± 1.92b 24.40 ± 1.54cd 27.90 ± 2.07bc 29.40 ± 1.93bc 22.60 ± 2.32d 30.11 ± 0.89a
HOS-G HNAN(%) Normal 4.30 ± 0.45c 6.60 ± 0.85bc 4.60 ± 0.65c 6.60 ± 1.24bc 7.80 ± 1.10b 9.10 ± 1.12b 8.60 ± 0.99b 13.50 ± 1.52a 7.64 ± 0.47b
Low Grade 13.40 ± 2.35c 12.10 ± 1.75c 22.80 ± 2.35ab 18.40 ± 2.04bc 23.00 ± 1.59ab 24.40 ± 2.78ab 14.40 ± 1.85c 28.60 ± 1.54a 19.63 ± 0.94a
SPD (mm/20 min) NormalNS 20.50 ± 0.45 19.10 ± 0.56 20.50 ± 0.43 20.00 ± 0.65 19.30 ± 0.30 20.30 ± 0.72 19.40 ± 0.48 18.90 ± 0.38 19.75 ± 0.19a
Low Grade 15.50 ± 0.69a 14.50 ± 0.43abc 15.20 ± 0.49ab 14.30 ± 0.26abc 13.30 ± 0.42c 13.90 ± 0.55bc 13.90 ± 0.31bc 14.60 ± 0.60abc 14.40 ± 0.19b

Table 1: HOS-G and sperm penetration distance of normal and low grade fresh semen (Means ± SE) in Ongole bulls.

HPAP, HPAN, HNAP and HNAN percentages differed significantly between normal and low grade ejaculates as well as between the bulls in both grades of ejaculates (Table 2). HPAP sperm percent of normal fresh semen had a highly significant negative correlation on HNAN sperm percent of fresh semen (P<0.01) and a significant positive correlation with live sperm percent and HPAP sperm percent of post diluted semen (P<0.05), while the HPAP sperm percent of fresh low grade semen had a highly significant positive correlation with HPAP sperm percent post diluted low grade semen (P<0.01).

Source of variation df Mean sum of squares with F value in parenthesis
HOS-G HPAP(%) HOS-G HPAN(%) HOS-G HNAP(%) HOS-G HNAN(%) SPD (mm/20 min)
Between groups 1 27352.900 (1000.824)** 189.225 (6.482)* 5187.006 (120.028)** 5760.000 (130.853)** 1144.900 (424.435)**
Error 158 27.330 29.195 43.215 44.019 2.697
Between groups-normal ejaculates 7 87.60 (6.62)** 66.714 (3.37)** 82.479 (4.66)** 85.684 (7.90)** 4.229 (1.61)NS
Error 72 13.233 19.483 17.675 10.843 2.631
Between groups-low grade ejaculates 7 174.857 (8.23)** 86.021 (2.88)** 393.727 (12.76)** 355.713 (8.30)** 5.171 (2.17)*
Error 72 21.225 29.831 30.860 42.840 2.375

Table 2: Analysis of variance for HOS-G and sperm penetration distance between normal and low grade fresh semen in Ongole bulls.

The overall mean sperm penetration distance was recorded as 19.75 ± 0.19 and 14.40 ± 0.19 mm/20 min in normal and low grade ejaculates (Table 1) with a highly significant difference between them (P<0.01). The SPD (mm/20 min) differed only amongst bulls of low grade ejaculates and not among those of normal ejaculates (Table 2). Sperm penetration distance of fresh semen had a significant negative correlation with abnormal sperm count (P<0.05), while the same for low grade semen revealed a significant positive correlation with post diluted HNAP sperm percent and post diluted sperm penetration distance (P<0.05). Sperm penetration distance of normal fresh semen had a significant positive correlation with fertility (P<0.05), while it had a non-significant negative correlation in low grade ejaculates.

Discussion

In the present study HOS positive and acrosome positive (HPAP) percent differed significantly between normal and low grade ejaculates (P<0.05) and had a highly significant difference between the bulls in both the grades of semen (P<0.01). HOS positive and acrosome negative (HPAN) percent differed non significantly between normal and low grade ejaculates and had a highly significant difference between the bulls in both the grades of semen (P<0.01) which was in agreement with the observations of Sushant and Kumar [8].

HPAP sperm percent of normal fresh semen had a non-significant positive correlation with fertility, while the HPAP sperm percent of low grade fresh semen had a non-significant negative correlation with fertility. Kumaresan et al. [9] opined that HOS test evaluates the biochemical intactness plasma membrane as normal, intact active sperm plasma membrane was essential for fertilization to occur.

The overall mean sperm penetration distance recorded in the present investigation was in corroboration with those of Kumar et al. [10] while Sushant and Kumar [8] reported higher sperm penetration distance values assessed for 60 minutes in Holstein Friesian bulls and their crosses and concluded that the values differed significantly between bulls and breeds as observed in the present study.

Sperm penetration distance of normal fresh semen had a significant negative correlation on abnormal sperm count of fresh semen (P<0.05) and had a significant positive correlation with fertility (P<0.05), while it had a non-significant negative correlation in low grade ejaculates as spermatozoal characteristics such as motility, morphology, concentration and acrosome integrity significantly affect the sperm penetration as opined by Anilkumar et al. [11].

It was concluded from the present study that routine semen evaluation tests which are mainly subjective in nature suffered human bias and have limited ability to predict the fertilizing capacity of the sperm, hence still more sensitive tests like functional assessment of plasma membrane and acrosome membrane (HOS-G Test) and sperm penetration distance in synthetic medium are essential to predict optimum fertility as reported by Kumaresan et al. [9]. These tests have been performed in the present investigation with ease and produced consistent results and hence could be adopted for routine semen evaluation as synthetic medium (2% polyacrylamide gel) could be stored in the refrigerator for prolonged use to evaluate sperm penetration distance as availability of bovine cervical mucus is restricted and could not be stored for prolonged usage.

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