Quality of ram spermatozoa separated with modified swim up method

2018 ◽  
Vol 33 (2) ◽  
pp. 62-70 ◽  
Author(s):  
A Hossain ◽  
MM Islam ◽  
F Naznin ◽  
RN Ferdousi ◽  
FY Bari ◽  
...  
Keyword(s):  
Semen Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Fluid Medium ◽  
Normal Morphology ◽  
The Mean ◽  
Mass Activity ◽  
Fresh Semen ◽  
Artificial Vagina

Semen was collected from four rams, using artificial vagina and viability%, motility% and plasma membrane integrity% were measured. Fresh ejaculates (n = 32) were separated by modified swim-up separation using modified human tubal fluid medium. Four fractions of supernatant were collected at 15-minute intervals. The mean volume, mass activity, concentration, motility%, viability%, normal morphology and membrane integrity% (HOST +ve) of fresh semen were 1.0 ± 0.14, 4.1 ± 0.1 × 109 spermatozoa/ml, 85.0 ± 1.3, 89.4 ± 1.0, 85.5 ± 0.7, 84.7 ± 0.5 respectively. There was no significant (P>0.05) difference in fresh semen quality parameters between rams. The motility%, viability% and HOST +ve % of first, second, third and fourth fractions were 53.4 ± 0.5, 68.2 ± 0.3, 74.8 ± 0.3 and 65.5 ± 0.4; 55.5 ± 0.4, 66.2 ± 0.4, 74.5 ± 0.3 and 73.6 ± 0.3 and 66.7 ± 0.5, 66.8 ± 0.5, 65.2 ± 0.4 and 74.7 ± 0.5 respectively. The motility%, viability% and membrane integrity% of separated semen samples differed significantly (P<0.05) between four fractions. The mean motility% and viability% were significantly higher (P<0.05) in third fraction (74.8 ± 0.3%), whereas the mean HOST +ve% was significantly higher (P<0.05) in fourth fraction (74.7 ± 0.5). All quality parameters of separated spermatozoa were significantly (P<0.05) lower than that of fresh semen. The pregnancy rates were higher with fresh semen (71%) in comparison to that of separated sample (57%).Bangl. vet. 2016. Vol. 33, No. 2, 62-70

scholarly journals Effect of preservation time on the quality of frozen semen in indigenous rams

2015 ◽  
Vol 44 (1) ◽  
pp. 10-15 ◽  
Author(s):  
BBA Mahmuda ◽  
Azizun Nesa ◽  
BF Zohara ◽  
MGS Alam ◽  
FY Bari
Keyword(s):  
Semen Quality ◽  
Egg Yolk ◽  
Volume Density ◽  
Normal Morphology ◽  
Mean Values ◽  
Frozen Semen ◽  
Significant Difference ◽  
The Mean ◽  
Fresh Semen

The study was carried out to observe the effects of preservation time on the quality of frozen semen of indigenous rams. Semen was collected using AV once a week from 4 rams. Tris based with 10% egg yolk and 7% glycerol extender was used to extend and freezing the semen. Fresh semen was evaluated for volume, density, mass motility and concentration, and mean values were observed as 0.8±0.2ml, 3.0±0.3, 3.2±0.7, 3.9±0.7×109/ml, respectively. Significant difference (p<0.05) was found for all the parameters among the rams. Mean values of motility, viability and normal morphology percentages were 83.3±4.3%, 88.2±4.4%, 84.2±3.5% in fresh semen while those of chilled semen at 40C were 74.7±2.3, 78.8±4.9 and 79.2±2.9%, respectively. For all the parameters, significant (p<0.05) difference was found among the rams. Frozen sperm motility was observed after thawing at 39-400C for 14-15 seconds. The mean motility, viability and normal morphology percentages after freezing for 24hrs, 7, 15 and 30 days of duration were 39.8±3.1, 41.1±4.3, 40.1±4.1 and 39.4±2.9%; 44.5±2.5, 45.3±2.8, 44.6±2.8 and 43.9±2.8%; 71.0±2.0, 71.7±1.5, 70.7±1.7 and 70.3±1.8%, respectively and values did not decrease significantly (p>0.05) with the increasing time of preservation. Non significantly decrease of the semen quality with advance of preservation time indicates the suitability of the protocol used for freezing of indigenous ram semen in Bangladesh.DOI: http://dx.doi.org/10.3329/bjas.v44i1.23113            Bang. J. Anim. Sci. 2014. 44 (1): 10-15

scholarly journals Spectrophotometric Analysis of the Resazurin Reduction Test as a Tool for Assessing Canine Semen Quality

2013 ◽  
Vol 57 (2) ◽  
pp. 281-285 ◽  
Author(s):  
Rafał Strzeżek ◽  
Krystyna Filipowicz ◽  
Marta Stańczak ◽  
Władysław Kordan
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Spectrophotometric Analysis ◽  
Normal Morphology ◽  
Additional Tool ◽  
Reduction Test ◽  
Highly Correlated

Abstract The resazurin reduction test (RRT) was subjected to spectrophotometric analysis to evaluate the quality of canine semen. Twenty four samples of canine semen were analysed. The absorption peaks for resazurin and resorufin were determined at 615 and 580 nm, respectively. The RRT ratio (RRTsperm-the ratio for samples containing spermatozoa, RRTplasma-the ratio for samples containing seminal plasma) was calculated by dividing the absorbance at 580 nm by the absorbance at 615 nm. Spearman’s correlation test was used to determine the significance of correlations between the analysed sperm parameters and the results of the resazurin reduction assay. The RRT ratio was highly correlated with sperm motility (r=0.68, P<0.01), progressive sperm motility (r=0.61, P<0.01), the subpopulation of cells with rapid velocity (r=0.72, P<0.01), and the subpopulation of cells with medium velocity (r= -0.54, P<0.05). A negative correlation was observed between the reducing capacity of seminal plasma vs. sperm with plasma membrane integrity (r= -0.60, P<0.01) and sperm with normal morphology (r= -0.58, P<0.01). The RRT test can be used as an additional tool for evaluation of the quality of canine semen.

17 EFFECT OF LOCAL TREATMENT OF SEMINAL VESICULITIS ON THE QUALITY OF EQUINE FRESH SEMEN

2015 ◽  
Vol 27 (1) ◽  
pp. 101
Keyword(s):  
Semen Quality ◽  
Local Treatment ◽  
Membrane Integrity ◽  
Ringer Lactate ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Seminal Parameters ◽  
Fresh Semen ◽  
Lactate Solution

Stallions affected by seminal vesiculitis present history of infertility or subfertility, ejaculatory disturbance, spread of sexually transmitted pathogens, and changes in semen characteristics, leading to reduced semen quality and longevity. The aim of this study was to evaluate the semen quality of stallions with seminal vesiculitis before and after local treatment. Five stallions with a mean age of 12.4 years diagnosed with seminal vesiculitis were used. The identification of the microorganism involved in the pathogenesis of seminal vesiculitis of each animal was performed by bacterial culture of the seminal vesicles flush with Ringer Lactate solution, performed in duplicate at 1-week intervals. After identification of bacteria was performed, there was susceptibility testing to antibiotic (antibiogram) and the appropriate antibiotic was chosen. The local treatment was performed by endoscopy for 10 consecutive days, and this consisted of flushing with Ringer Lactate solution, followed by infusion of the antibiotic selected. The semen analyses were performed before starting the local treatment for seminal vesiculitis (M0), after a week (M1), and after a month (M2) of therapy. Sperm kinetics were performed by computerized method – CASA for the following parameters: percentage of sperm with total motility, progressive motility, and rapid sperm. Analysis of plasma membrane integrity was performed by epi-fluorescence microscopy, using the combination of fluorescent probes carboxyfluorescein diacetate and propidium iodide. Percentage of leukocytes was assessed through evaluation in light optical microscopy of semen smears stained with DiffQuick. The content of nitric oxide (NO) was determined by colourimetric Griess reaction by a spectrophotometer through the concentrations of nitrate (NO3–) and nitrite (NO2–). To perform the count of colony forming units per millilitre (CFU mL–1), an aliquot of 0.1 mL of semen was diluted in 9.9 mL of saline. A 0.1-mL aliquot of this sample was plated on Mueller-Hinton agar. The seeded plates were incubated, and the bacterial colonies were counted after 24 h. According to the performed dilution, total colonies identified corresponds to ×10 000 CFU mL–1. The data were analysed by two-way ANOVA followed by Tukey's test (P < 0.05). The values (mean ± standard error) of seminal parameters on M0, M1, and M2 were the following, respectively: sperm kinetics (total motility: 46.5 ± 5.13a; 75.1 ± 3.42b; 42.8 ± 5.28a; progressive motility: 19.3 ± 3.86a; 33.4 ± 2.39b; 16.5 ± 2.40a; rapid sperm: 22.2 ± 1.82a; 52.2 ± 5.65b; 22.1 ± 2.62a); plasma membrane integrity (47.5 ± 4.65a; 62.9 ± 5.41b; 39.1 ± 4.32a); percentage of leukocytes (35.2 ± 2.36a; 15.1 ± 2.55b; 36.1 ± 4.04a); CFU (119 980 × 103 ± 19 528.0 × 103a; 5375 × 103 ± 2453.7 × 103b; 65 850 × 103 ± 19 701.0 × 103ab) on fresh semen; and NO content (0.645 ± 0.172a, 0.117 ± 0.023b, 0.364 ± 0.110ab) on seminal plasma. The results demonstrate that local treatment after a week leads to an improvement in sperm quality; however, this was not maintained after 1 month of therapy, since the seminal parameters at this time are similar to pretreatment, which can be justified by recurrent disease.

scholarly journals Effect of adding Manganese chloride and Co-enzymes (α-lipoic acid and Q10) on post-cryopreservation semen quality characteristics of Holstein bulls

2018 ◽  
Vol 41 (2) ◽  
pp. 86-93
Author(s):  
Eidan S. M.
Keyword(s):  
Lipoic Acid ◽  
Semen Quality ◽  
Manganese Chloride ◽  
Control Group ◽  
Control Groups ◽  
Holstein Bulls ◽  
Animal Resource ◽  
Fresh Semen ◽  
Artificial Vagina

     This study was undertaken to explore the adding effect of manganese chloride (MnCl2), co-enzyme Q10 (Co-Q10) as well as α-lipoic acid to Tris extender on Freeze ability of Holstein bulls’ semen. This study was carried out at the department of artificial insemination at the Directorate of Animal Resource, Ministry of Agriculture in Abu-Ghraib, Baghdad for the duration from October\ 2013 to Jun\ 2014, including three experiments. Seven Holstein bulls of 3.5-4 years old were used in this study. Semen was collected via artificial vagina by one ejaculate/ bull/ week. The assessments were conducted for fresh semen, which was later pooled, equally divided for various treatments within each experiment, using Tris extender. In the first experiment, pooled semen was divided into three groups. First group was diluted with Tris only. Manganese chloride was added to Tris extender (0.7 mM) in the 2nd group while in 3rd group (0.9 mM) of the Manganese chloride was used. In the second experiment, semen was divided equally into three groups. The first group was considered as a control group diluted with Tris only. Co-enzyme Q10 was added to 2nd (0.2 mM) and 3rd groups (0.5 mM) treatments respectively. In the third experiment, semen was divided into three groups. The first group was diluted with Tris only (control group). While the 2nd and 3rd groups were added 0.5 and 1.0 mM α-lipoic acid respectively. The effect of these additions on Holstein bulls semen quality was studied during different periods (48 hours, one, two and three months post cryopreservation) for three experiments. The results revealed that the addition two levels of the MnCl2 (2nd and 3rd, experiment 1), Co- Q10  (2nd and 3rd, experiment 2) and α- lipoic acid (2nd and 3rd experiment 3) led to significant increases freezability as compared with control groups during all the experiment periods. In conclusion, the addition of MnCl2, Co-Q10 and α-lipoic acid led to improved post-cryopreservation semen quality of Holstein bulls. This will in turn enhance fertility rate of artificially-inseminated cows and owners economic income consequently.

scholarly journals The effect of addition selected amino acids in extender semen on quality and DNA stability of frozen-thawed Sumba Ongole bull spermatozoa

2019 ◽  
Vol 24 (1) ◽  
Author(s):  
Syahruddin Said ◽  
Setiorini Setiorini ◽  
Amaitshaa Adella ◽  
Indah Sari ◽  
Nursafira Fathaniah ◽  
...  
Keyword(s):  
Amino Acids ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Dna Integrity ◽  
Freezing And Thawing ◽  
Dna Stability ◽  
Cattle Breeding ◽  
Egg Yolks

<p class="abstrak1"><span lang="IN">The objective of the current study was to asses the optimal concentration of glutamine, glycine and cysteine amino acids in tris-citric-acid-fructose egg yolks (TCFY) extender on quality of SO bull spermatozoa during freezing and thawing. </span><span lang="EN-US">In t</span><span lang="IN">his study the DNA stability of frozen-thawed Sperm</span><span lang="EN-US"> was </span><span lang="IN">also indentified. Three mature bulls maintained at PT. Karya Anugerah Rumpin, private cattle breeding company, West Java, Indonesia were used as semen donors. Semen was collected using artificial vagina and were evaluated </span><span lang="EN-US">prior</span><span lang="IN"> to freezing. Semen was diluted with TCFY supplemented with different concentrations of amino acids (5, 15 and 25 mM glycine and glutamine, and 3, 5 and 7 mM cysteine) then processed for colling and freezing. Semen quality parameters (subjective motility, viability and membrane and DNA integrity). </span><span lang="EN-US">D</span><span lang="IN">ata showed that in general the effect of addition of selected amino acids (glycine, glutamine and cysteine) </span><span lang="EN-US">in</span><span lang="IN">to TCFY extenders on motility, viability and membrane integrity of SO spermatozoa after cooling were significantly different (p&lt;0.05) higher than</span><span lang="EN-US"> that of</span><span lang="IN"> control. Addition of 15 mM glycine, 15 mM glutamine and 5 mM cysteine resulted in significant (p&lt;0.05) increase post-thawing sperm motility and sperm viability as compared to th</span><span lang="EN-US">at of</span><span lang="IN"> control. Furthermore, when spermatozoa were stained with acridine orange after fixation with acetic alcohol, the DNA integrity of post-thawing spermatozoa showed that all spermatozoa were remain intact. In conclusion </span><span lang="EN-US">,</span><span lang="IN">addition of 15 mM glycine, glutamine and 5 mM cysteine increase the cryoprotecting efficacy of bovine bull cryopreservation extender, and furthermore all DNA spermatozoa were remain intact. </span></p>

scholarly journals Comparative effects of addition of superoxide dismutase and reduced glutathione on cryopreservation of Sahiwal bull semen

2019 ◽  
Vol 69 (4) ◽  
pp. 1291
Author(s):  
A. Murtaza ◽  
M. Ahmad ◽  
M. Zubair ◽  
S. Umar ◽  
A. Mushtaq ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Semen Quality ◽  
Reduced Glutathione ◽  
Membrane Integrity ◽  
Bull Semen ◽  
Microscopic Evaluation ◽  
Artificial Vagina

The present study aimed to investigate effects of superoxide dismutase (SOD) and reduced glutathione (GSH) on the quality of frozen-thawed semen of Sahiwal bulls. Semen was collected twice a week for 8 weeks by artificial vagina from six Sahiwal bulls, kept at the Semen Production Unit Qadirabad, Sahiwal-Pakistan. After gross and microscopic evaluation, qualifying semen ejaculates were divided into 10 equal aliquots and diluted in extenders enriched with no antioxidants (control); or supplemented with either SOD (50, 100 and 200 IU/mL), or GSH (0.5, 1 and 2 mM) or their combinations (50 IU/mL SOD and 0.5 mM GSH, 100 IU/mL SOD and 1 mM GSH and 200 IU/mL SOD and 2 mM GSH). Samples were then frozen and stored in liquid nitrogen at -196°C for 24 h. The following parameters were evaluated for semen quality: post-thawed sperm motility, viability, acrosome and membrane integrity. According to the results, sperm motility, viability, acrosome and membrane integrity were significantly (P<0.05) higher in samples treated either with 100 IU/mL of SOD; 1 mM and 2 mM of GSH or 50 IU/mL of SOD plus 0.5 mM of GSH. In conclusion, semen quality might be improved by supplementing semen extenders with 100 IU/mL of SOD; 0.5 and 1 mM of GSH and combination of 50 IU/mL and 0.5 mM of SOD and GSH, respectively.

scholarly journals DAYA HIDUP SPERMATOZOA KAMBING PERANAKAN ETAWA YANG DIPRESERVASI DENGAN BERBAGAI JENIS PENGENCER

2016 ◽  
Vol 3 (3) ◽  
pp. 81
Author(s):  
Muhammad Rizal ◽  
Muhammad Thahir
Keyword(s):  
Artificial Insemination ◽  
Sodium Citrate ◽  
Randomized Design ◽  
Completely Randomized Design ◽  
Mass Activity ◽  
Fresh Semen ◽  
Artificial Vagina

The purpose of this research was to examine the effectivity of Tris, sodium citrate, and lactose extenders in maintaining viability of etawa crossbreed (EC) goat spermatozoa preserved at 3–5oC.  Semen was collected using artificial vagina once a week for six times as replicate.  Fresh semen was divided into three tubes then diluted with Tris, sodium citrate, and lactose extenders, respectively.  Diluted-semen were stored in refrigerator at 3–5oC.  Quality of diluted-semen including percentages of motile spermatozoa (MS) and live spermatozoa (LS) were evaluated every day during storage at 3–5oC for four days.  Data were analyzed using completely randomized design with three treatments and six replicates.  Results of this study showed that mean volume, colour, consistency, pH, mass activity, spermatozoa concentration, MS, LS, and spermatozoa abnormal of PE goat fresh semen were 1.05 ml, cream, thick, 6.9, ++ – +++, 3,756.67 million cell/ml, 76.67%, 85.83%, and 4.5%, respectively.  Tris, sodium citrate, and lactose extenders have same ability in maintaining viability of EC goat spermatozoa during preserved at 3–5oC.  At day-4 storage, percentages of MS and LS for Tris (44.17 and 57.83%) was no difference significant with sodium citrate (43.33 and 56.67%), and lactose (43.33 and 54.17%).   In conclusion, Tris, sodium citrate, and lactose extenders have same ability in maintaining viability of EC goat spermatozoa during preserved at 3–5oC.  Tris, sodium citrate, and lactose extenders could be maintaining 40% in percentage of motile spermatozoa for three days after preserved at 3–5oC, and suitable for use in artificial insemination (AI) programKeywords: Tris, sodium citrate, lactose, spermatozoa viability, PE goat.

scholarly journals Comparative Study of Gir Cattle and Surti Buffalo Bulls Semen under Middle Gujarat Climate

2017 ◽  
Vol 13 (01) ◽  
Author(s):  
P. J. Chaudhary ◽  
A. J. Dhami ◽  
D. V. Chaudhari ◽  
K. K. Hadiya ◽  
J. A. Patel
Keyword(s):  
Sperm Concentration ◽  
Membrane Integrity ◽  
Mean Values ◽  
Abnormal Sperm ◽  
Buffalo Semen ◽  
The Mean ◽  
Mass Activity ◽  
Live Sperm ◽  
Surti Buffalo ◽  
Fresh Semen

This study was undertaken during the favourable breeding season of the year 2016-17 on healthy mature Gir cattle and Surti buffalo bulls, three each, at Sperm Station of the College. The ejaculates (9/bull, total 54) collected in the morning using artificial vagina were evaluated for routine seminal attributes, including acrosomal and plasma membrane integrity. The mean values of ejaculate volume, sperm concentration, mass activity (0-5 score), individual sperm motility, live sperm, abnormal sperm, intact acrosome and HOST reactive sperms observed in fresh semen of Gir cattle and Surti buffalo bulls were 6.20±0.42 and 3.34±0.23 ml (P less than 0.01), 1169.44±61.71 and 846.30±54.82 million (P less than 0.01), 3.44±0.09 and 3.42±0.08, 76.53±0.53 and 80.76±0.39 % (P less than 0.05), 81.00±1.32 and 84.73±0.78 % (P less than 0.05), 6.00±0.37 and 5.81±0.40 %, 95.59±0.35 and 95.54±0.25 % as well as 80.30±1.90 and 84.58±0.88 % (P less than 0.05), respectively. The variation between cattle and buffalo semen was significant for most of these traits. The variations between bulls within breed were however not significant. Significant correlations were observed only between mass activity and initial motility (0.62), and live and abnormal sperm (-0.41) in Gir bulls, and for ejaculate volume with sperm concentration (-0.56) and abnormal sperm (0.45), and between live and HOS reactive sperms (0.48) in Surti bulls.

scholarly journals PENGARUH METODE PENCUCIAN SPERMATOZOA SAPI ACEH TERHADAP MOTILITAS, PERSENTASE HIDUP, DAN INTEGRITAS MEMBRAN PLASMA UTUH SPERMATOZOA (The Infuence of Aceh Bull Spermatozoa Washing Method on Spermatozoa Motility and Plasma Membrane Integrity of Intact Spermatozoa)

2015 ◽  
Vol 9 (2) ◽  
Author(s):  
Listin Handayani ◽  
Dasrul Dasrul ◽  
Muslim Akmal ◽  
Cut Nila Thasmi ◽  
Hamdan Hamdan ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Spermatozoa Motility ◽  
Sperm Washing ◽  
Fresh Semen

This study aimed to determine the effect of sperm washing by swim up and centrifugation in isotonic medium on sperm quality of aceh bull. In this study, fresh semen from healthy male aceh bull aged 3-4 months was collected using artificial vagina. Immediately after semen collection, fresh semen quality was examined macroscopically and microscopically. Subsequently, sperm washing was performed by centrifugation and swim up in sperm washing medium. Group 1 (P0) as control group, cement washed with isotonic solution (andromed medium: saline solution) with ratio of 1:8. 2. Group 2 (P1), cement was separated by centrifugation method, group 3 (P2), all cement was separated by swim up method then examined the sperm quality sperm washing results. Each treatment was repeated 5 times. Quality parameters measured were the percentage of spermatozoa motility, sperm viability, and plasma membrane integrity intact spermatozoa. Data were analyzed with analysis of variance one-way pattern, followed by Duncan's multiple test. The results showed the mean ± SD percentage of sperm motility of each treatment group (P0; P1; P2) respectively amounted to 72.00±3.74, 66.40±4.77, and 73.60±3.29%. The percentage of viability was 72.00 ±3.74%, 66.40±2.88%, 71.80±2.17%. The percentage of plasma membrane integrity is intact spermatozoa was 68.20±1.79%, 57.20±3.77%, 69.00±2.00%. Results of this study showed that the percentage of motility, live spermatozoa and plasma membrane integrity intact after separation by swim-up method were significantly different (P <0.05) compared with no separation.Key words: spermatozoa quality, aceh bulls, centrifugation, swim up

Bacterial contamination of ram semen used for artificial insemination in indigenous ewes

2018 ◽  
Vol 34 (1) ◽  
pp. 20-26 ◽  
Author(s):  
E Ahmed ◽  
MS Islam ◽  
MGS Alam ◽  
PK Jha ◽  
S Ghosh ◽  
...  
Keyword(s):  
Bacterial Contamination ◽  
Biochemical Tests ◽  
E Coli ◽  
Bacillus Spp ◽  
The Mean ◽  
Mass Activity ◽  
Staphylococcus Spp ◽  
Fresh Semen ◽  
Artificial Vagina ◽  
Growth Of Bacteria

Ram semen was assessed for quality and presence of bacteria. Four ejaculates were collected from each of four rams twice a week using artificial vagina. The volume varied from 0.4 - 1.3 mL, colour from 2 - 4 (creamy to creamy-grey), mass activity from 3 - 5, sperm motility from 75 - 85%, viability from 80 - 95%, and concentration from, 2500 - 5000 × 106/mL. The mass activity of ram R6 was significantly (P<0.05) higher (5.0 ± 0.0) compared with ram R1 (4.4 ± 0.5), R2 (3.9 ± 0.0) and R5 (4.7 ± 0.5). The mean motility was 81.7 ± 4.0, viability 90.0 ± 4.0 and concentration 3519.0 ± 545.6 x 106/ml. E. coli and Staphylococcus spp. were found in all four rams’ fresh semen confirmed by culture, staining and biochemical tests. However, Bacillus spp. was found only in ram R5. When the semen samples were treated with antibiotics there was no growth of bacteria after three days of incubation. It is suggested antibiotics control the transmission of microorganisms through AI in ewes.Bangl. vet. 2017. Vol. 34, No. 1, 20-26

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