Effects of dietary supplementation of bentonite and Saccharomyces cerevisiae cell wall on acute-phase protein and liver function in high-producing dairy cows during transition period.

The aim of this study was to investigate the efficacy of dietary endotoxin binders [bentonite (BEN) and Saccharomyces cerevisiae cell wall (SCW)] on acute-phase protein (APP) response and liver function in cows during the transition period. Twenty-four multiparous Holstein cows were randomly assigned to one of four treatment groups. The experimental groups consisted of (1) the basal diet (BD) + SCW, (2) BD + SCW + BEN, (3) BD + BEN, and (4) BD (control). Blood samples were taken at 1, 3 and 4 weeks before and 1 and 3 weeks after parturition and serum concentrations of non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), glucose, haptoglobin (Hp), serum amyloid A(SAA), albumin, g-glutamyl transferase (GGT), aspartate aminotransferase (AST), cholesterol, iron, and lipopolysaccharide (LPS) were measured. The concentrations of LPS, SAA, albumin, and Hp in the blood were within reference range at all times. The level of blood LPS was not high enough to initiate an APP response. Mean BHBA concentration was highest at 1 week after calving. For NEFA, the pattern was similar, with a peak at 1 week after calving. Cholesterol concentration was lower in the SCW group, probably due to a lower lipoprotein concentration. Mean AST concentration was highest at 1 week after calving, especially in the SCW + BEN group. The results of a current study showed that, if the carbohydrate level is not high in the diet to cause rumen acidosis, it is not profitable to supplement BEN and SCW for adsorbing endotoxins in the diet, in transition cows.

Altering undigested neutral detergent fiber through additives applied in corn, whole barley crop, and alfalfa silages, and its effect on performance of lactating Holstein dairy cows.

OBJECTIVE: We hypothesized that silage additives may alter the undigested neutral detergent fiber (uNDF) content through ensiling. Therefore, urea and formic acid were applied to corn, whole barley crop (WBC) and alfalfa to change uNDF content of the ensiled forages. METHODS: Six experimental diets at two groups of high uNDF (untreated corn and alfalfa silages [CSAS] and untreated whole barley and alfalfa silages [BSAS]) and low uNDF (urea-treated corn silage+untreated alfalfa silage [CSUAS], urea-treated whole barley silage+untreated alfalfa silage [BSUAS], untreated corn silage+formic acid-treated alfalfa silage [CSASF], and untreated whole barley silage+formic acid-treated alfalfa silage [BSASF]), were allocated to thirty-six multiparous lactating Holstein dairy cows. RESULTS: The untreated silages were higher in uNDF than additive treated silages, but the uNDF concentrations among silages were variable (corn silage0.05). Milk yield tended to increase in the cows fed high uNDF diets than those fed low uNDF (p = 0.10). The cows fed diet based on urea-treated corn silage had higher milk yield than those fed other silages (p = 0.05). The substitution of corn silage with the WBC silage tended to decrease milk production (p = 0.07). Changing the physical source of NDF supply and the uNDF content from the corn silage to the WBC silage caused a significant increase in ruminal NH3-N concentration, milk urea-N and fat yield (p< 0.05). The cows fed diets based on WBC silage experienced greater rumination time than the cows fed corn silage (p<0.05). CONCLUSION: Administering additives to silages to reduce uNDF may improve the performance of Holstein dairy cows.

WITHDRAWN:The Effect of Intermittent Watering on Performance, Blood Metabolites, and Nutrient Digestibility in Baluchi Lambs.

Objective: This study was conducted to evaluate the effect of intermittent watering on Baluchi lambs, regarding their performance, blood metabolites, and nutrient digestibility. Methods: Twenty male Baluchi lambs, 120 ± 5 days of age and 29 ±2.2 kg body weight were randomly assigned to four watering treatments: (T1) free access to water during the day, (T2) watered ad libitum twice a day, (T3) watered ad libitum once a day, (T4) watered ad libitum once every two days. The experiment lasted 42 days. Results: water restriction led to reduction in weight gain, DMI, water consumption, Water/DMI (p<0.05). Rectal temperature and respiration rate non influenced (p>0.05). aldosterone, cortisol, insulin hormone affected by treatments (p<0.05). Serum glucose, creatinine, total protein and globulin not affected (p> 0.05). while observed significant difference among T3 with other treatment in BUN (p<0.05). significant differences were observed in serum cholesterol and triglyceride between T4 and other groups (p< 0.05). There was a significant difference between T4 and T1 and T2 in albumin serum (p< 0.05). RBC WBC non influenced by water restriction (p< 0.05). However, PCV, MCV, MCH, MCHC and hemoglobin influenced by treatments (p< 0.05). sodium and chloride of serum was significantly higher in T4, and there were significant differences among the treatments (p< 0.05). But there were no differences among the treatments (p> 0.05), with regard to calcium, phosphorus, magnesium, potassium. Digestibility of ADF, NDF, CP and OM was different among T3, T4 and other treatments (p< 0.05), while no differences were found in EE digestibility among treatments (p> 0.05). There was a significant difference between T4 and other groups and between T4 and T1 regarding fecal output and urine exertion respectively. Conclusion: This results implies that Baluchi lambs can tolerate water restriction up to once a day for at least 28 days without harmful effects.

Effect of micronization and meal size of corn grain on glycemic response and in vitro hindgut acidosis potential in horses.

This study aimed to 1) evaluate the interaction of corn grain micronization and starch levels per meal on equine plasma glucose, and 2) determine if micronization affects the risk of hindgut acidosis. Six mature (aged 6 to 10 years), healthy, non-pregnant mares (initial body weight [BW]: 301 to 463 kg) were used in a 2×3 factorial cross-over design. The treatments included two forms of corn grain (ground and micronized flaked) at three levels of starch (1, 1.5, and 2 g/kg BW per meal). The blood was sampled before and 30, 60, 90, 120, 180, 240, and 300 min after morning feeding and the glucose concentration in the plasma was determined. Small intestine and hindgut dry matter (DM) disappearances of ground and micronized corn were also compared using in vitro techniques. Micronized flaked corn grain showed three times more in vitro enzymatic DM disappearance (p < 0.001) compared with ground corn. Residues of in vitro enzymatic digestion of micronized flaked corn fermented 38.59 % faster than ground corn during in vitro hindgut incubation. The horses that consumed micronized flaked corn had higher post-prandial plasma glucose concentrations (p < 0.001). Increasing starch levels per meal from 1-2 g/kg BW resulted in higher plasma glucose concentrations (P = 0.005). However, no interaction of processing and starch meal size was found. Overall, processing the corn grain by micronization or increasing starch level per meal increased the plasma glucose concentrations, but the magnitude of the increases did not match that expected from in vitro studies.

Protein molecular structures in alfalfa hay cut at three stages of maturity and in the afternoon and morning and relationship with nutrient availability in ruminants.

BACKGROUND: Molecular structures in feed protein influence its digestive behavior, availability and utilization. From a nutritive point of view, stage of maturity and cutting time are important factors affecting nutrient profiles and availability of alfalfa (Medicago sativa L.) hay in ruminants. The objectives of this study were to determine protein molecular structures by Fourier transform infrared spectroscopy (FTIR), and their relationship with nutrient profiles and availability in ruminants of alfalfa hay cut at early bud, late bud and early flower stages and in afternoon and morning. RESULTS: With advancing maturity, molecular structure ratios of α-helix:ß-sheets decreased, while amide I:amide II increased (P ≤ 0.05). Alfalfa cutting in afternoon versus morning increased protein structure α-helix:ß-sheets and α-helix:others ratios (P < 0.05) and tended to decrease the proportion of ß-sheets (P = 0.09). Positive correlations were found for α-helix:ß-sheet ratio (R ≥ 0.60; P < 0.05) with intermediately degradable protein (PB2) and ruminal degradability and intestinal protein supply, and all these parameters correlated negatively with amide I:amide II ratio (R ≤ -0.62; P < 0.05). CONCLUSION: Protein molecular structures in alfalfa hay changed with advancing maturity and during the day and these protein structures affected predicted nutrient availability of alfalfa hay in ruminants.

Effects of monensin on metabolism and production in dairy saanen goats in periparturient period.

This trial evaluated the effects of dietary supplementation with monensin sodium on dry matter intake, metabolic parameters and milk yield and milk composition of dairy Saanen goats in the periparturient period. Twelve Saanen pregnant dairy goats were assigned to a treatment and a control group. Saanen goats were fed monensin as its 10% sodium salt in the amount of 33 mg/kg of total dry matter intake during 30 d before parturition till d 42 in milk. Blood samples were collected from each goat at d 30, 15 and 7 before expected kidding time and also in d 1, 7, 13, 19, 21, 28, 35 and 42 postpartum. The serum concentrations of ß-Hydroxybutyrate (BHBA), non-esterifed fatty acid (NEFA), calcium, magnesium, inorganic phosphorus, cholesterol, triglyceride, urea, total protein, albumin and glucose and aspartate aminotransferase (AST) activity were determined. The BHBA concentration significantly decreased in goats, which received monensin in comparison to controls in the postpartum period (p = 0.049). The concentration of sodium (Na) was significantly influenced by monensin treatment in the postpartum period (p = 0.048). In addition, the difference in dry matter intake was highly significant between the two groups during the pre-partum period. Controls had more dry matter intake (DMI) than treatment goats (p = 0.0001). Mean 3.5% fat corrected milk production was not influenced by monensin treatment. However, milk fat percentage was significantly decreased in monensin treated goats (p = 0.0017).

Determining Optimal Nonlinear Regression Models for Studying the Kinetics of Fatty Acid Ruminal Biohydrogenation In Vitro.

The accurate estimation of in vitro ruminal biohydrogenation (BH) kinetics of fatty acids (FA) allows for a more accurate understanding of their dynamics and develop targeted strategies to enhance desirable FA bypass. This study comprises a comprehensive evaluation of 33 nonlinear regression models to determine the most suitable model for accurately estimating the in vitro BH kinetics of individual FA. The data set utilized in the present research originates from a recent investigation on the effects of micronization and vitamin E on the in vitro ruminal BH of rapeseed. For the nonlinear regression analysis, data comprising FA concentrations (expressed as g FA/100 g FA) at the conclusion of 2, 4, 8, 12, 24, and 48 h incubation periods were employed. The evaluation of nonlinear regression models focused on identifying the ideal model based on criteria including the highest R2 value, the lowest RMSE value, and statistically significant coefficients. The results pinpoint the Gompertz model as an effective choice for estimating the in vitro ruminal BH kinetics of upward-trending fatty acids, including intermediate unsaturated fatty acids and saturated end FA. Additionally, the first-order kinetic model of Ørskov and McDonald emerges as the preferred model for investigating the BH kinetics of downward-trending fatty acids, including oleic acid, linoleic acid, and alpha-linolenic acid. In summary, this rigorous evaluation led to the identification of the most appropriate model, one that not only exhibited an exceptional fit to the data but also provided profound insights into the intricate relationships between predictors and the dynamic behavior of FA. The established nonlinear regression models will serve as invaluable tools for future research investigating FA biohydrogenation kinetics.

Influence of flaxseed with rumen undegradable protein level on milk yield, milk fatty acids and blood metabolites in transition ewes.

An experiment was conducted to determine the effects of two levels of rumen undegradable protein (RUP) without or with whole or extruded flaxseed on milk yield, milk component, milk fatty acids (FAs) profile and plasma metabolites in transition ewes. Three weeks before and after lambing, seventy-two Baluchi ewes were used in a completely randomized design with a 3 × 2 factorial arrangement of treatments. The treatments contained 1) no flaxseed + 20% RUP (no flaxseed, low RUP [NFLR]); 2) no flaxseed + 40% RUP (no flaxseed, high RUP [NFHR]); 3) 10% whole flaxseed + 20% RUP (whole flaxseed, low RUP [WFLR]); 4) 10% whole flaxseed + 40% RUP (whole flaxseed, high RUP [WFHR]); 5) 10% extruded flaxseed + 20% RUP (extruded flaxseed, low RUP [EFLR]), and 6) 10% extruded flaxseed + 40% RUP (extruded flaxseed, high RUP [EFHR]). Ewes fed 10% extruded flaxseed exhibited higher (p < 0.001) dry matter intake (DMI) and colostrum yield (p < 0.1) compared to other treatments. Two types of flaxseed and RUP levels had no significant effect on milk yield, but milk fat and protein contents decreased and increased in diets containing 40% RUP, respectively. Ewes fed extruded flaxseed produced milk with lower concentrations of saturated fatty acids (SFA) and higher α-linolenic and linoleic acids and also polyunsaturated fatty acids (PUFA) compared to other groups (p < 0.05). During post-lambing, the ewes fed diets containing flaxseed exhibited higher concentration of serum non-esterified FAs (NEFA) compared to diets without flaxseed (p < 0.01). The concentration of serum ß-hydroxybutyric acid (BHBA) decreased in the diets containing flaxseed types at pre-lambing, but increased in diets containing extruded flaxseed at post-lambing (p < 0.01). The serum glucose concentration of ewes (pre and post-lambing) which consumed diets containing extruded flaxseed or 40% RUP increased, but blood urea concentration was elevated following supplementation of diet with whole flaxseed or 40% RUP (p < 0.001). In conclusion, utilization of 10% extruded flaxseed in the diets of transition ewes had positive effects on animal performance with favorable changes in milk FAs profile. However, there is no considerable advantage to supply more than 20% RUP level in the diet of transition dairy sheep.

Metabolic Fingerprinting of Feces from Calves, Subjected to Gram-Negative Bacterial Endotoxin.

Gram-negative bacteria have a well-known impact on the disease state of neonatal calves and their mortality. This study was the first to implement untargeted metabolomics on calves' fecal samples to unravel the effect of Gram-negative bacterial endotoxin lipopolysaccharide (LPS). In this context, calves were challenged with LPS and administered with fish oil, nanocurcumin, or dexamethasone to evaluate treatment effects. Ultra-high-performance liquid-chromatography high-resolution mass spectrometry (UHPLC-HRMS) was employed to map fecal metabolic fingerprints from the various groups before and after LPS challenge. Based on the generated fingerprints, including 9650 unique feature ions, significant separation according to LPS group was achieved through orthogonal partial least squares discriminant analysis (Q2 of 0.57 and p-value of 0.022), which allowed the selection of 37 metabolites as bacterial endotoxin markers. Tentative identification of these markers suggested that the majority belonged to the subclass of the carboxylic acid derivatives-amino acids, peptides, and analogs-and fatty amides, with these subclasses playing a role in the metabolism of steroids, histidine, glutamate, and folate. Biological interpretations supported the revealed markers' potential to aid in disease diagnosis, whereas beneficial effects were observed following dexamethasone, fish oil, and nanocurcumin treatment.

Different processing effects on nutritive value of barely grains by gas production method.

The purpose of this study was to investigate the effects of warm and cold physical as well as chemical processing methods on the nutritive value of barley grains by gas production technique. The processing methods included milling, steam flaking, extruding and soaking up the grain in water containing 1.00% citric acid, 1.00% propionic acid and 0.01 M sulfuric acid. Two-hundred mg of milling samples were incubated in special 100 mL glasses and the amount of gas produced at different hr was measured. The data were analyzed in a completely randomized design. The results showed that different treatments did not have a significant effect on chemical composition of barley seeds. Other methods of processing compared to the grinding method, significantly reduced the total amount of produced gas. In the 72 hr incubation period, the lowest amount of gas production was in the extruded (245.6 mL per g dry matter) treatment. However, there was no significant difference between the two methods of extruding and flaking. The highest percentage of digestible organic matter was associated with propionic acid (64.90%), while the steam cracking method (58.74%) was the least. Among the processing methods, the highest amounts of methane production, total protozoa population and volatile fatty acid concentration were related to the grinding method and the least amount of extrusion treatment was observed during 24 hr of incubation. Different experimental treatments had a significant effect on ammonia nitrogen condensation and the highest level was observed in milling. According to our results, processing methods such as extrusion and flaking may improve the grain nutritive value.

Effects of feeding rumen-protected amino acids on the performance of feedlot calves.

OBJECTIVE: This study was conducted to produce and evaluate protected amino acids (AAs) against degradation in the rumen with greater bioavalibility and without the problems associated with polymer coating and the effect this has on calf performance. MATERIALS AND METHODS: In the first step, essential AAs methionine and lysine were reacted with two chemical compounds (Benzaldehyde and Glutaraldehyde) in an attempt to make ligands for producing protected AAs. The physico-chemical characterization, melting point, and mass spectrometric of products were estimated. These products were fed to 36 Holstein dairy calves with 110 ± 0.50 kg of average body weight and an age of 110 ± 10 days. Calves were randomly assigned to six treatments. This study was done with six treatments as a completely randomized one-way design. RESULTS: Feed consumption and average daily gain were less for control animals and those fed methionine and lysine glutaraldehyde compared to other treatments. The largest chewing time was observed for methionine and lysine glutaraldehyde, respectively, and the least was control. There was no difference for energy consumption, dry matter intake, or blood metabolites among the six treatments. The greatest total protein content was related to methionine and lysine glutaraldehyde treatment and the least total protein was observed in control treatment. CONCLUSION: It can be concluded that the use of chemical methods to protect AAs can be applied and may have some beneficial effects.

Effects of phosalone consumption via feeding with or without sodium bentonite on performance, blood metabolites and its transition to milk of Iranian Baluchi sheep.

BACKGROUND: Transfer of pesticides from environment to animal products is inevitable, so the purpose of the present work was to evaluate phosalone consumption via feeding with or without sodium bentonite (SB) on performance, blood metabolites and its transition to milk of Iranian Baluchi sheep. METHODS: Twenty Baluchi ewes were divided into four treatments (P1 as control, P2, P3, and P4) of five animals in which phosalone, an organophosphate pesticide, was given via diet (only for P2 and P3) at a dose of 280 mg/sheep/day for 63 consecutive days. The SB (32 g/sheep/day; for P3 and P4) was also evaluated for its ability to reduce deleterious effects of phosalone in the sheep diets. The control group (P1) did not receive any phosalone and SB during the experiment. Sampling was conducted in two periods of time including weeks 5 and 9. RESULTS: Phosalone residues were observed in the milk samples of P2 and P3 groups during two sampling periods. During period 1, the transfer rate of phosalone from feed to milk was 0.23 and 0.02%, respectively for the contaminated diets (P2 and P3), which is relatively similar to period 2 (0.22 and 0.02%). Only 0.34 (period 1) and 0.36% (period 2) of phosalone residue are excreted in the feces of P2 group following its daily consumption. Transfer of phosalone from feed to milk was affected (P < 0.05) by the dietary inclusion of a commercial SB, as it (SB) decreased excretion of phosalone via milk (P3). The phosalone and SB alone or together had no significant effect (P > 0.05) on the dry matter intake (DMI) and body weight (BW) gain, but feed efficiency, milk production, milk fat, dry matter (DM) and organic matter (OM) digestibility, acetylcholinesterase (AChE) inhibitory activity, hemoglobin (Hb), red blood cell (RBC), serum glutamic pyruvic transaminase (SGPT), serum glutamic oxaloacetic transaminase (SGOT), albumin and mean corpuscular hemoglobin concentration (MCHC) affected by the treatments in period 1 or 2 (P < 0.05). The Hb, RBC, and MCHC were significantly decreased (P < 0.05) by about 9.72, 20.77, and 9.71%, respectively in the group P2 as compared to those of the control group during period 1. The AChE inhibitory activity (period 1 and 2) significantly increased when phosalone administered via the diet (P < 0.05). CONCLUSIONS: Although there were no adverse effects on the performance of sheep following the intake of phosalone alone (P2 vs. P1), but other research on the long and short times to the phosalone in high and low doses with more animals is suggested. Overall, compared to the control group, addition of SB in the diet of sheep improved nutrient digestibility, animal performance, and milk health.

Evaluation of SCD and FASN Gene Expression in Baluchi, Iran-Black, and Arman Sheep.

BACKGROUND: With the increasing concern for health and nutrition, dietary fat has attracted considerable attention. The composition of fatty acids in the diet is important because they are associated with major diseases including cancers, diabetes, and cardiovascular disease. The fatty acid synthase (FASN) and stearoyl-CoA desaturase (delta-9-desaturase) (SCD) genes affect fatty acid composition (1). The expression of SCD and FASN genes is related to an increase in conjugated linoleic acid (CLA) in dairy products, which benefits human health.The aim of current study was to investigate expression changes of SCD and FASN genes that resulted from crossbreeding the local Baluchi sheep with alien breeds. METHODS: We collected tissue samples from the mammary glands of 24 single-born ewes from local Baluchi and synthetic Iran-Black and Arman sheep breeds in the Abbas Abad breeding center. After RNA extraction and cDNA synthesis, real-time PCR was performed with all samples in triplicate. RESULTS: The maximum and minimum expression of SCD and FASN genes was in the local Baluchi sheep and the crossbred Arman sheep, respectively. CONCLUSION: With the highest SCD and FASN gene expression in local Baluchi sheep and relatively less expression of these genes in synthetic Iran-Black and Arman Sheep breeds, it may be necessary to consider the consequences of crossbreeding local sheep and the fatty acid composition of their dairy products.

Impacts of feeding selenium-methionine and chromium-methionine on performance, serum components, antioxidant status, and physiological responses to transportation stress of Baluchi ewe lambs.

The effects of selenium-methionine (Se-Met) and chromium-methionine (Cr-Met) supplementation on performance and response to transportation stress were studied on 24 Baluchi ewe lambs (18-20 weeks of age) for 9 weeks. The lambs were randomly assigned to four dietary treatments: (1) control; (2) 1.5 mg supplemental Se-Met/kg dry matter (DM) of diet; (3) 0.8 mg supplemental Cr-Met/kg DM of diet; and (4) 1.5 mg Se-Met plus 0.8 mg Cr-Met/kg DM of diet (Se-Cr-Met). At the commencement of week 8, a road transportation stress (TS) was carried out for 30 min. Lambs fed Cr-Met and Se-Cr-Met diets had higher feed intake than the control and Se-Met animals (P < 0.0001). Lambs on Cr-Met diet showed higher average daily gain (ADG) compared to the control group (P = 0.007). Se-Met and Cr-Met supplementation alone or in combination significantly (P < 0.05) reduced feed conversion ratio (FCR). The animals that received Se-Met (P = 0.014), Cr-Met (P = 0.005), and Se-Cr-Met (P = 0.003) supplemented diets had lower glucose concentration than the control. Lambs on Cr-Met had higher blood T3 concentration than control animals (P = 0.040), while Cr-Met (P = 0.039) and Se-Cr-Met (P = 0.032) supplementation increased triiodothyronine (T3) to thyroxin (T4) ratio. Animals fed Se-Met and/or Cr-Met supplements had lower blood malondialdehyde (MDA) in week 9 of the experiment (P < 0.05). Blood ferric-reducing antioxidant power (FRAP) tended to be higher in the Se-Met- and Se-Cr-Met-supplemented groups (P < 0.1).TS reduced feed intake in lambs fed the control diet in week 8 of the experiment (P = 0.003). The lambs given with supplemental Cr-Met exhibited lower glucose concentration before transportation (BT) (P = 0.029) and after transportation (AT) (P = 0.016) compared to the control. Lambs fed Se-Cr-Met had the lowest cortisol concentration BT (P < 0.05). It was concluded that feeding Se-Met and/or Cr-Met supplements could improve growth performance and be beneficial in attenuating the adverse effects of transportation stress in Baluchi ewe lambs.

An evolutionary relationship between Stearoyl-CoA Desaturase (SCD) protein sequences involved in fatty acid metabolism.

BACKGROUND: Stearoyl-CoA desaturase (SCD) is a key enzyme that converts saturated fatty acids (SFAs) to monounsaturated fatty acids (MUFAs) in fat biosynthesis. Despite being crucial for interpreting SCDs' roles across species, the evolutionary relationship of SCD proteins across species has yet to be elucidated. This study aims to present this evolutionary relationship based on amino acid sequences. METHODS: Using Multiple Sequence Alignment (MSA) and phylogenetic construction methods, a hypothetical evolutionary relationship was generated between the stearoyl-CoA desaturase (SCD) protein sequences between 18 different species. RESULTS: SCD protein sequences from Homo sapiens, Pan troglodytes (chimpanzee), and Pongo abelii (orangutan) have the lowest genetic distances of 0.006 of the 18 species studied. Capra hircus (goat) and Ovis aries (Sheep) had the next lowest genetic distance of 0.023. These farm animals are 99.987% identical at the amino acid level. CONCLUSIONS: The SCD proteins are conserved in these 18 species, and their evolutionary relationships are similar.

Quality of bovine chilled or frozen-thawed semen after addition of omega-3 Fatty acids supplementation to extender.

BACKGROUND: This study was conducted to evaluate the potential protective effects of omega-3 poly unsaturated fatty acids (Ω-3 PUFAs) on bovine sperm quality in response to cooling and cryopreservation. MATERIALS AND METHODS: : In this experimental study included ejaculates from five proven fertile bulls, allocated to the control and the four experimental groups. For group 1, polyethylene glycol (PEG) as a solvent was added alone to the extender, while for groups 2, 3 and 4, different concentration of omega-3 PUFAs (1, 2.5 and 5%, respectively) in combination with PEG were added to the semen extender. Motility [using computer aided sperm analysis (CASA)], viability and morphology of bovine sperm were investigated after 24 and 48 hours in both cold liquid storage and frozen-thawed conditions. RESULTS: Our findings showed that PEG has some detrimental effects on sperm quality. Cooling as well as cryopreservation decreased significantly most of measured variables of sperm as compared to fresh semen, whereas the treatments did not improve sperm quality. Furthermore, levels of some variables were decreased significantly during treatments (p<0.05). CONCLUSION: Addition of Ω-3 PUFAs to semen extenders cannot be effectively introduced to conservation media as well as sperm membrane in order to protect spermatozoa in response to cooling and freezing. It can be suggested if Ω-3 PUFAs is supplemented to the diet of bulls in order to modify the fatty acid compositions of sperm, they might perform their preventive properties.