Bacteriospermia and Sperm Quality of Cryopreserved Bull Semen Used in Artificial Insemination of Cows in South Wollo Zone, Ethiopia

The objectives of this trial were to estimate prevalence of bacteriospermia, to determine the bacterial load, and to isolate the types of bacteria as well as to assess the association between bacterial load and sperm quality traits in cryopreserved bull semen in field conditions in the South Wollo Zone. A total of 309 cryopreserved straws of semen from the Holstein Friesian (HF)-cross bull (n = 180 straws) and pure Jersey bull (n = 129 straws) were investigated. Bacteriological assessments of the presence of aerobic bacteria, estimation of bacterial count and bacterial isolation, as well as semen quality were performed. Aerobic bacterial contamination was prevalent in 38.8% of the semen straws. No significant difference in the prevalence of bacteriospermia was observed among bulls although the HF-cross bull had a higher prevalence (40.0%). But, significant difference in prevalence of bacteriospermia was found among semen ejaculates of the same bull. The risk of bacteriospermia in the HF-cross bull was higher (Odds ratio = 1.86, 95% CI = 0.168–20.26) compared to Jersey although not significant. Overall average bacterial load of 50.38 ± 16.29 colony-forming units (CFU)/ml (from nil to 1318.20 CFU/ml) was found. No significant difference in bacterial count among bulls and their ejaculates was observed. Moreover, correlation analysis revealed that the proportions of motility, live, and normal morphology were negatively influenced by an increase in the bacterial contamination of semen. In this study, three isolates of coagualse-negative Staphylococcus species and one isolate of Corynebacterium species were found. Average percentages of sperm motility (48.35 ± 1.23), live (66.08 ± 1.0), and normal morphology (80.62 ± 1.24) were observed. It was concluded that cryopreservation does not guarantee the quality of semen from bacterial contamination. Hence, meticulous care should be adopted to prevent contamination of semen by bacteria during collection, transportation, processing, and storage times.

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Effect of whole and clarified egg yolk based extenders on post-thaw semen quality in bulls.

10.31220/agrirxiv.2021.00050 ◽

2021 ◽

Author(s):

Prosper Kamusasa ◽

Eddington Gororo ◽

Fungayi Primrose Chatiza

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

Egg Yolk ◽

Quality Parameters ◽

Normal Morphology ◽

Semen Cryopreservation ◽

Bull Semen ◽

Inclusion Level ◽

Whole Egg

Abstract This study was conducted to evaluate the comparative cryoprotective effects of whole egg yolk and clarified egg yolk on post thaw sperm quality parameters and to determine the optimum clarified egg yolk inclusion level (10-20%) in semen extenders for Mashona bull semen cryopreservation. It was shown that there was a significant decrease in sperm quality variables following cryopreservation. Semen quality increased with the concentration of clarified egg yolk, indicating a positive relationship between egg yolk LDL concentration and maintenance of in vitro sperm quality. The 20% clarified egg yolk (CEY20) extender treatment gave post-thaw motility, viability and normal morphology values which were comparable to the control (20% whole egg yolk, WEY20). The 10% clarified egg yolk concentration gave the least post-thaw quality values and the greatest proportion of defective spermatozoa. This experiment found no advantage of replacing whole egg yolk with up to 15% clarified egg yolk in Mashona bull semen cryopreservation. However, 20% clarified and 20% whole egg yolk performed similarly in the maintenance of post-thaw sperm motility, viability and normal morphology.

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Efficacy of potassium permanganate and turmeric as antimicrobial agents on the bacterial load and quality of boar semen during preservation at 15 °C

Veterinarski arhiv ◽

10.24099/vet.arhiv.0715 ◽

2020 ◽

Vol 90 (5) ◽

pp. 443-451

Author(s):

Nang Kanna Manpoong ◽

Kutubuddin Ahmed ◽

Dilip Kumar Bhattacharya ◽

Dipak Kumar Sarma ◽

Nekibuddin Ahmed ◽

...

Keyword(s):

Antimicrobial Agents ◽

Semen Quality ◽

Sperm Quality ◽

Bacterial Load ◽

Conception Rate ◽

E Coli ◽

Bacillus Spp ◽

Boar Semen ◽

Conventional Antibiotics

The present study was conducted to evaluate the efficacy of different natural antimicrobials agents (KMnO4 and Turmeric) in comparison with conventional antibiotics against the bacterial load and in relation to the quality of boar semen in Modena extender for up to 120 hours of preservation at 15 °C. A total of 56 ejaculates, 14 from each of four Hampshire crossbred boars maintained within the ICAR-AICRP on Pigs, in Guwahati, Assam, India, were utilized in the study. Thirty-two ejaculates, 8 from each of four boars were used to study the effect of antimicrobial agents on semen quality during preservation at 15 °C in Modena extender. A total of 9 different bacterial types were identified from 46 bacterial isolates, obtained from 24 fresh semen samples viz. Staphylococcus aureus (24%), E. coli (22%), Bacillus spp. (13%), Citrobacter spp. (9%), Pseudomonas spp. (9%), Staphylococcus epidermidis (9%), Klebsiella spp. (6%), Streptococcus spp. (6%) and Proteus spp. (2%). The overall sensitivity of the recovered isolates to Gentamicin, Ampicillin, Enrofloxacin, Cloxacillin, Streptomycin, Penicillin, Amoxycilln, Ofloxacin and Tetracyclin were 89, 39, 37, 48, 74, 52, 56, 76 and 63% respectively. The mean sperm motility, intact acrosome, HOST-reacted spermatozoa and bacterial load differed significantly (P˂0.01) between antimicrobial agents (Gentamicin, KMnO4 and Turmeric) and preservation periods (0, 48, 72, 96 and 120 hours). Sperm quality based on Gentamicin was found to be best, followed by Turmeric and KMnO4 during preservation at 15 °C. The conception rate for the semen preserved for 0, 24, 48, 72, 96 and 120 hours of preservation was 83.33, 80.00, 75.00, 66.66, 66.66 and 50.00% respectively. In the present study, the preserved semen with ascending bacterial load containing Gentamicin did not affect the conception rate.

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

Bangladesh Journal of Animal Science ◽

10.3329/bjas.v44i1.23113 ◽

2015 ◽

Vol 44 (1) ◽

pp. 10-15 ◽

Cited By ~ 1

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

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Practical use of bioenergetic evaluation of sire-bulls with different quality of semen production

BIO Web of Conferences ◽

10.1051/bioconf/20213204003 ◽

2021 ◽

Vol 32 ◽

pp. 04003

Author(s):

Ludmila Samusenko ◽

Andrey Mamaev ◽

Natalya Rodina ◽

Ekaterina Sergeeva

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

The Body ◽

Biologically Active ◽

Black And White ◽

Field Of Vision ◽

Short Period ◽

Significant Difference ◽

Active Centres

For modern cattle industry, it is very important to evaluate bull-sires of different breeds by semen quality. The relevance of this problem arises in the context of development of artificial insemination technologies and biotechnology, the increasing role of sire-bulls in the genetic improvement of the herd, which makes it possible to significantly change the breed composition of herds in large regions in a short period of time. In cattle breeding, sires of Black-and-white and Holstein black-and-white breeds, which are imported into the Russian Federation, are used almost throughout the country. The purpose of the study is to assess the practical possibility of using the measured data of the bioelectric potential of superficially localized biologically active centres of sire-bulls to evaluate the quality of their semen production. The subject of the study was the semen production of the bulls of Black-and-white and Holstein blackand-white breeds. The groups were formed according to the principle of analogues; the bulls were from 2 to 4 years old; the body weight was from 700 to 800 kg. As a result of evaluating the sperm quality in sire-bulls with a high level of biopotential of the superficially localized biologically active centres, in the field of vision of the preparation, spermatozoa with an intact acrosome were found, which amounted to 96.5% of the total number of spermatozoa of the preparation and had a significant difference with the experimental group. The use of bioenergetic data of the superficially localized biologically active centres makes it possible to evaluate the sperm quality before semen collection and thereby, reduce the time and labour costs for low-quality ejaculates culling.

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The Efficiency of Selected Extenders against Bacterial Contamination of Boar Semen in a Swine Breeding Facility in Western Slovakia

Animals ◽

10.3390/ani11113320 ◽

2021 ◽

Vol 11 (11) ◽

pp. 3320

Author(s):

Eva Tvrdá ◽

Ondřej Bučko ◽

Kristína Rojková ◽

Michal Ďuračka ◽

Simona Kunová ◽

...

Keyword(s):

Bacterial Contamination ◽

Sperm Quality ◽

Bacterial Species ◽

Bacterial Load ◽

Bacterial Count ◽

Total Bacterial Count ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Gram Positive Bacteria ◽

Boar Semen

Bacteriospermia has become a serious factor affecting sperm quality in swine breeding, this is why antibiotics (ATBs) are a critical component of semen extenders. Due to ever-increasing antimicrobial resistance, the aim of this study was to assess the efficiency of selected commercially available semen extenders to prevent a possible bacterial contamination of boar ejaculates. Three Androstar Plus extenders containing different combinations of antibiotics were used to process ejaculates from 30 healthy Duroc breeding boars. Androstar Plus without antibiotics was used as a control. The extended samples were stored at 17 °C for 72 h. Sperm motility, viability, mitochondrial activity, DNA integrity and oxidative profile of each extended sample were assessed following 24 h, 48 h and 72 h. Furthermore, selective media were used to quantify the bacterial load and specific bacterial species were identified with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The results indicate that semen extenders enriched with ATBs ensured a significantly higher preservation of the sperm quality in comparison to the ATB-free control. The total bacterial count was significantly decreased in the extenders supplemented with ATBs (p < 0.001), however gentamycin alone was not effective enough against Gram-positive bacteria, while a few colonies of Enterococcus hirae, Bacillus subtilis and Corynebacterium spp. were present in the samples extended in the presence of a triple combination of ATBs. In conclusion, we may suggest that semen extenders enriched in antibiotics were not able to fully eliminate the bacteria present in the studied samples. Furthermore, selection of suitable antibiotics for semen extension should be accompanied by adequate hygiene standards during the collection and handling of boar ejaculates.

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Kualitas spermatozoa semen sexing kambing peranakan etawa (pe) dengan metode sedimentasi putih telur menggunakan pengencer yang berbeda

Jurnal Sains Peternakan ◽

10.21067/jsp.v5i1.3136 ◽

2017 ◽

Vol 5 (1) ◽

pp. 39-49

Author(s):

Kornelis Tamo Ama ◽

Enike Dwi Kusumawati ◽

Aju Tjatur Nugroho Krisnaningsih

Keyword(s):

Semen Quality ◽

Sperm Quality ◽

Egg White ◽

Laboratory Research ◽

Sedimentation Method ◽

Randomized Design ◽

Significant Difference ◽

Completely Randomized Design ◽

Fresh Semen

ABSTRAK Tujuan penelitian ini adalah untuk mengetahui kualitas spermatozoa semen hasil sexing kambing Peranakan Etawa (PE) dengan metode sedimentasi putih telur menggunakan pengencer yang berbeda. Materi yang digunakan dalam penelitian ini adalah semen segar kambing PE. Metode percobaan yang digunakan adalah percobaan laboratorium dengan menggunakan rancangan acak lengkap (RAL) Setiap perlakuan semen sexing kambing Peranakan Etawa dengan pengencer yang berbeda yaitu (P0) CEP, (P1) CEP+kuning telur+putih telur, (P2)CEP+putih telur (P3), CEP+kuning telur dan diulang sebanyak 10 kali. Variabel yang diukur pada penelitian ini adalah motilitas, viabilitas, dan abnormalitas spermatozoa. Data yang diperoleh dianalisis dengan mengunakan analisis varian. Apabila perlakuan memberikan pengaruh maka dilanjutkan dengan uji Beda Nyata Terkecil (BNT). Hasil penelitian menunjukkan bahwa kualitas spermatozoa kambing Peranakan Etawa (PE) dengan berbagai macam pengencer menunjukkan pengaruh yang sangat nyata (P<0,01). Motilitas dan Viabilitas menunjukkan perbedaan yang sangat nyata (P<0,01) pada berbagai pengencer Motilitas terbaik pada beberapa lapisan atas sebesar 65% dan CEP+PT lapisan atas 60% dilanjutkan oleh CEP+KT+PT lapisan bawah 55,9% dan CEP+PT lapisan bawah 55% dilanjutkan lagi oleh CEP+KT lapisan atas 50% dan lapisan bawah 45% dan yang paling terkecil adalah CEP lapisan bawah 40%. Viabilitas spermatozoa dari yang tertinggi yaitu CEP+KT+PT lapisan atas sebesar 69,553% dan CEP+PT lapisan atas sebesar 69,519% dan dilanjutkan dengan CEP+PT lapisan bawah sebesar 65,504% dan CEP+KT+PT lapisan bawah 65,473%, dan dilanjutkan lagi dengan CEP+KT lapisan atas sebesar 60,269% dan lapisan bawah sebesar 53,476% dan yang paling terkecil yaitu CEP lapisan bawah sebesar 40,371%. Presentase Abnormalitas menunjukkan bahwa perlakuan tidak memberikan pengaruh nyata (P>0,05). Tetapi CEP+KT+PT lapisan atas menunjukkan persentase yang paling rendah yaitu sebesar 4,88% dibandingkan perlakuan lainnya. Dari hasil penelitian dapat disimpulkan bahwa kualitas spermatozoa semen sexing kambing PE dengan menggunakan pengencer CEP+ kuning telur + putih telur pada lapisan atas memberikan hasil terbaik terhadap kualitas spermatozoa ditinjau dari motilitas, viabilitas dan abnormalitas. ABSTRACT The purpose of this study was to determine the sexing sperm quality of Etawa cross-bred goat (PE) with egg white sedimentation method using different diluents. The material used in this study was the fresh semen Etawa crossbreed goat (PE) of the center for Artificial Insemination (BBIB) Singosari Malang. The method is laboratory research using completely randomized design. Consist of CEP, CEP + KT, CEP + PT, CEP + KT + PT and repeated 10 times. The variables are motility, viability and abnormality of sperm. Data were analyzed by using variance analysis. If the treatment effect then continued by Least Significant Difference (LSD). The results showed that the quality of Etawa cross-bred goat sperm with various diluents showed a significant influence (P <0.01). Motility and viability showed differences (P <0.01) in various diluents. The best motility and viability on top layer of CEP+KT+PT diluent were 65% and 69%, 55%. Percentage abnormalities showed that the treatment was not significant effect (P> 0.05). but top layer CEP+KT+PT diluent shows the percentage of abnormality west 4.88% compared to the other treatments. From the results of this study concluded that the quality of sexing semen quality by using dilution CEP + yolk white egg on the top layer gives the best results on the quality of sperm in terms of motility, viability and abnormalities. Based on this study it is suggested that the use of sexing semen with egg white sedimentation method using diluent CEP + yolk + white egg.

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Hygiene assessment of sheep slaughter cycle

Bulletin of the Veterinary Institute in Pulawy ◽

10.2478/bvip-2014-0036 ◽

2014 ◽

Vol 58 (2) ◽

pp. 243-246

Author(s):

Renata Pyz-Łukasik ◽

Waldemar Paszkiewicz

Keyword(s):

Bacterial Contamination ◽

Bacterial Load ◽

Bacterial Count ◽

Stage I ◽

Stage Iii ◽

Total Bacterial Count ◽

Microbiological Analysis ◽

Mean Values ◽

Significant Difference ◽

Commission Regulation

Abstract The purpose of the study was to determine microbial contamination of mutton carcass surface with regard to the number of the slaughtered animals. The total bacterial load and Enterobacteriaceae and enterococci counts were determined. Sampling for microbiological analysis as well as detection and enumeration of each microorganism group were performed according to the Polish Standards. No significant effect of the order of the slaughtering animals during the slaughter day on total bacterial count on mutton carcass surfaces was found. The overall bacterial contamination of mutton carcasses were between 1.0 × 103 cfu/cm2 (3.0 log - stage I) and 2.5 × 103 cfu/cm2 (3.4 log - stage III). No significant difference among the slaughter cycles, as indicated by total microbial numbers was observed. The obtained daily mean log values ranged from 4.7 × 102 (2.67 log) and 7.6 × 103 (3.88 log) cfu/cm2. The daily log mean values were lower than the maximal bacteria count (M) set out for hygiene standard of sheep slaughter process by the Commission Regulation 2073/2005. Bacteria belonging to the Enterobacteriaceae family were recovered from 21 (65.6%) samples while enterococci were identified in 28 (87.5%) samples. In most cases, significant differences in the level of contamination with bacteria isolated from the carcasses at each stage of a daily slaughter cycle, were not observed. At stage III, significantly higher levels of bacterial contamination (0.86 and 1.31 log cfu/cm2 respectively) were established as compared to stage I (0.37 and 0.58 log cfu/cm2 respectively). There were no Salmonella-positive samples determined. Importantly, the number of slaughtered animals during a slaughter day did not influence bacterial contamination on carcass surface if the successful application of HACCP control system was combined with the implementation of optimal sanitary supervision.

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Quality of ram spermatozoa separated with modified swim up method

Bangladesh Veterinarian ◽

10.3329/bvet.v33i2.36459 ◽

2018 ◽

Vol 33 (2) ◽

pp. 62-70 ◽

Cited By ~ 2

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

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Macro- and microelements in serum and seminal plasma as biomarkers for bull sperm cryotolerance

Acta Veterinaria Scandinavica ◽

10.1186/s13028-021-00590-2 ◽

2021 ◽

Vol 63 (1) ◽

Author(s):

Maja Zakošek Pipan ◽

Petra Zrimšek ◽

Breda Jakovac Strajn ◽

Katarina Pavšič Vrtač ◽

Tanja Knific ◽

...

Keyword(s):

Seminal Plasma ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Characteristics ◽

Freezing And Thawing ◽

Additional Tool ◽

Bull Semen ◽

Bull Sperm ◽

Fresh Semen

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.

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Effect of docosahexanoic acid on quality of frozen–thawed bull semen in BioXcell extender

Reproduction Fertility and Development ◽

10.1071/rd15089 ◽

2017 ◽

Vol 29 (3) ◽

pp. 490 ◽

Cited By ~ 3

Author(s):

Asmatullah Kaka ◽

Wahid Haron ◽

Rosnina Yusoff ◽

Nurhusien Yimer ◽

A. M. Khumran ◽

...

Keyword(s):

Lipid Peroxidation ◽

Liquid Nitrogen ◽

Sperm Motility ◽

Membrane Integrity ◽

Optimum Level ◽

Docosahexanoic Acid ◽

Normal Morphology ◽

Bull Semen ◽

Acrosome Integrity

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen–thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15 ng mL–1 DHA was added. The supplemented semen samples were incubated at 37°C for 15 min for DHA uptake by spermatozoa. Later, samples were cooled for 2 h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24 h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3 ng mL–1 significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3 ng mL–1 concentration of DHA resulted in superior quality of frozen–thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.

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