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Effect of Diluents and Preservation on Time Native Cock Semen

Das SK
Veterinary Surgeon
Department of Livestock Services, Dhaka, Bangladesh.

Rahman MM
Department of Surgery and Obstetrics, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Bhuiyan MMU
Department of Surgery and Obstetrics, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Begum N
Veterinary Surgeon
Department of Livestock Services, Dhaka, Bangladesh.

FY Bari
Department of Surgery and Obstetrics, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh.

Abstract/ Executive Summary:

Evaluation, dilution and preservation are the essential part of quality determination of native cock semen. Total of 21 semen samples were collected by abdominal massage from 7 native cocks at 3 days interval. Fresh semen was examined macroscopically for physical parameters. Most of the samples were found in yellowish white in color. The mean±SD volume, motility, concentration, dead and morphologically abnormal spermatozoa of semen samples varied from 0.3±0.01 to 0.5±0.03 ml, 58.3±2.9 to 71.7±3.5%, 3733±153 to 6067±130 x 10⁶/ml, 22.3±4.51 to 33.7±1.53% and 18.3±7.8 to 30.7±1.5%, respectively. Cock # 1 showed the highest body weight (1.3 kg) and produced significantly (P<0.05) highest ejaculate volume and motility among the cocks. However, significantly (P<0.05) higher concentration of spermatozoa was observed in cock # 5 compared with others. The most frequent sperm abnormalities recorded in the mid-piece (8.5 to 13.0 %), followed by the head (3.5 to 6.0%) and tail (1.5 to 6.0%) of spermatozoa. After diluted in Lake A and Lake B (antibiotic added) diluents and preserved at 4ºC for two days, semen samples were examined on Day 0 (15 min chilled), Day 1 and Day 2, respectively. With the advancement of preservation, sperm motility, live spermatozoa and sperm morphology deteriorated progressively in both the diluents. However, significantly (P<0.05) higher percentage of motility was observed on Day 0 (60.6±2.4%) compared with Day 2 (39.2±2.0%) in Lake A diluent only. it was observed that Lake B diluent showed significantly (P<0.05) higher number of motile spermatozoa than Lake A on Day 1 and 2. Similar with the motility, significantly (P<0.05) higher proportion of live spermatozoa was present in Lake A on Day 0 compared with Day 2. However, proportion of live spermatozoa was significantly higher in Lake B than Lake A on Day 2 only. Significantly higher (P<0.05) number of tail abnormality was present on Day 2 compared with Day 0 and 1 in both diluents. The overall abnormalities were found higher when diluted in Lake A than Lake B. The semen quality can be affected by the absence of antibiotic in the diluents and various preservation times.

Keywords: Cock, Semen, Diluents, Preservation

Location: Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh

Start Date: 00-07-2010

End Date: 00-11-2010

Program Area: Postharvest and Agro-processing

Commodity: Poultry

Objectives:

1. To evaluate the native cock semen immediately after collection,

2. To determine the proper diluents and storage time and morphological evaluation of cock semen for betterment of artificial insemination in poultry industry throughout the country.

Methodology:

The study was conducted in the Department of Surgery and Obstetrics, Bangladesh Agricultural University, Mymensingh during the period from July 2010 to November 2010. Apparently healthy seven (n= 7) mature local cocks were purchased from the various local markets of the Mymensingh district devoid of any external anatomical deformities. The average age and body weight of cocks were 34±1.15 weeks and 1.14±0.13 kg respectively. Each bird was kept in individual large cages. They were dewormed against digestive and gastrointestinal round worm by using Avinex ® powder (Renata Limited, Bangladesh) and vaccinated against Ranikhet diseases using intramuscular injection of RDV (LRI, Mohakhali, Bangladesh). Each bird was fed with 120 gm commercial Layer Layer 1 (Quality Feeds Limited, Bangladesh) mash diet daily, containing 2900 kcal/kg ME and 19% CP. Total feeds were given in two splits per day. Water was provided ad-libitum. During the four weeks of pre-experimental period, cocks were trained twice daily for semen collection by the massage technique. Each cock was ejaculated three times at three days interval between 9.00 to 10.00 AM throughout the experimental period. The cocks were taken gently from the cage and manipulated immediately. Semen was collected by abdominal massage technique (Lake, 1957) followed by manipulation of cloaca. The ejaculated volume was collected by 1ml disposable syringe. To minimize the stress and maximize the quality of semen, collection was always performed by the same operators, at the same time, and under the same conditions. Special care was taken to avoid contamination of semen with urine and transparent fluid, which could degrade the semen quality. Within 30 min. of collection, semen samples were divided into 3 parts. One part was kept for fresh semen evaluation viz; mass activity, motility, concentration, viability and morphology. Other two parts were diluted in to 1:4 ratio (1 part of semen: 4 part of extender) at room temperature with 2 different extenders viz: Lake A and Lake B and was placed into 10 ml glass tubes and stored at 4°C in a refrigerator for further evaluation. The evaluation was done for the effects of two different semen extenders on the motility, viability and sperm morphology on immediately after addition of diluents and Day 1 and Day 2 hrs of preservation. Several parameters were evaluated to characterize each cock’s fresh semen quality and effects of diluents and preservation periods. These were semen volume, color, mass activity, motility, concentration, viability and morphology. The concentration of spermatozoa was determined by using hemocytometer technique as described by Bane. A drop of (10µl) diluted semen (1:100) was placed on the edge of cover slip and spermatozoa were allowed to settle for five minutes on the hemocytometer before placing on the stage of microscope. The spermatozoa were then counted in five large squares; four at corners and one at the center of 25 large squares. The numbers of spermatozoa in five large squares were counted and average number was recorded. The concentration of the spermatozoa per ml of semen was calculated by multiplying the average number of spermatozoa in large five squares with dilution factor and expressed as 10⁶ million per ml. The mean and standard deviation (SD) of ejaculate volume, color, mass activity, motility, concentration, dead sperm and sperm with abnormal head, mid-piece and tail were measured by using descriptive statistics in Microsoft Office Excel work sheet 2007. The percentage of motility, live sperm, morphologically abnormal spermatozoa and the proportion of different (Head, Mid-piece and Tail) abnormalities were calculated as pooled data. The data recorded were compiled, tabulated and analyzed in accordance with the objective of the study by one way analysis of variance (ANOVA) and Duncan’s multiple range test (DMRT) using MSTATC ® software. The difference between values was considered significant when the P value was less than 0.05.

Source of Information: International Journal of Veterinary Science, 5(2): 51-57, P-ISSN: 2304-3075; E-ISSN: 2305-4360

Article Link (Online Journal): www.ijvets.com

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

On the basis of experiment concluded that the quality of native cock semen depends on various factor body weight, nutrition and housing; Cock having heavier body weight produced higher ejaculate volume and diluents with antibiotic was preferable as dilution for keeping at 4ºC for two days.

Knowledge Management: Journal