Roasted Flaxseed and Hydro-cooling Synergy: Impacts on Seminal Quality of Crossbred Bulls under Heat Stress

Roasted Flaxseed and Hydro-cooling Synergy: Impacts on Seminal Quality of Crossbred Bulls under Heat Stress | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Roasted Flaxseed and Hydro-cooling Synergy: Impacts on Seminal Quality of Crossbred Bulls under Heat Stress Samir Humeida Ali, C Kotresh Prasad, Pawan Singh, Mukesh Bhakat, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8156401/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract This study evaluated the effects of roasted flaxseed supplementation and surface water misting on semen quality and sexual behaviour of crossbred Karan Fries bulls during the summer season under tropical Indian conditions. A total of eighteen healthy Karan Fries bulls were randomly assigned to three experimental groups (n = 6 per group). Bulls in the control group (T1) received a standard ration without any supplementation. In the second group (T2), were supplemented with roasted flaxseed ( 1 kg/bull/day), while those in the third group (T3) received the same level of flaxseed supplementation along with periodic freshwater misting over the body surface. Physiological responses including body weight, rectal temperature, respiration rate, pulse rate, and circulating concentrations of testosterone and luteinizing hormone were monitored at fortnightly intervals over two consecutive periods. Sexual behaviour traits were also recorded at the same frequency. Semen was collected fortnightly and evaluated for ejaculate volume, colour, mass activity, sperm motility, concentration, viability, hypo-osmotic swelling test (HOST) response, and acrosomal integrity in fresh samples. Statistical analysis using one-way ANOVA revealed significantly higher rectal temperature, respiration rate, and pulse rate in bulls maintained under the control group, followed by those in T2 and T3 (P < 0.05). In contrast, bulls in T3 exhibited significantly reduced reaction time, dismounting time, and ejaculation time compared to the control group (P < 0.05). Measures of sexual performance, including libido score and mating ability, were significantly superior in T3 animals. Semen quality parameters such as ejaculate volume, sperm concentration, motility, livability, percentage of normal spermatozoa, HOST response, and acrosomal integrity were also highest in T3, followed by T2 and T1 (P < 0.05). Overall, the findings indicate that the combined application of roasted flaxseed supplementation and freshwater misting effectively alleviates heat stress, enhances sexual behaviour, and improves semen quality in Karan Fries bulls during the summer season in tropical environments. Libido flaxseed misting semen quality Karan Fries bull 1. Introduction The success of the crossbred program depends on the sound bull's ability to produce quality semen and effective spermatozoa, which are essential for producing high-quality germplasm through natural mating with cows or artificial insemination using liquid and/or frozen semen (Kumar et al., 2015). Reproductive efficiency of crossbred bulls is a key determinant of the success of artificial insemination programs in the cattle industry (Diskin, 2018). Several factors influence semen quality, including diet, management practices, climatic conditions, and genetic composition (Qureshi, 2004; Rehman et al., 2014). In males, the current intensification of heat stress scenarios may become a primary factor affecting bull fertility, as environmental temperature alters spermatogenesis and thus influences the reproductive process (Casu et al., 1991). Nutrition plays a central role in reproduction by modulating gamete development, ovulation, fertilization, and metabolite- and hormone-mediated reproductive processes (Wathes et al., 2007). Flaxseed is one of the most concentrated plant-based sources of omega-3 fatty acids, particularly alpha-linolenic acid (ALA) (Gutte et al., 2015). Flaxseed oil contains approximately 58% linolenic acid and provides potent antioxidant activity that supports animal health and physiological functions (Mourvaki et al., 2010). Flaxseed (Linum usitatissimum), a member of the family Linaceae and commonly referred to as linseed, is a rich source of alpha-linolenic acid. Dietary inclusion of flaxseed has been reported to elevate tissue levels of alpha-linolenic acid and eicosapentaenoic acid, fatty acids that play an important role in the synthesis of key reproductive hormones (Petit et al., 2004). Omega-3 polyunsaturated fatty acids, particularly docosahexaenoic acid (DHA), play a critical role in maintaining sperm membrane integrity and viability, thereby enhancing semen quality and resilience to stress in cattle (Gulliver et al., 2012). Flaxseed, as a rich dietary source of n-3 fatty acids, has been widely reported to alter sperm lipid composition and improve semen quality across species, including boars (Rooke et al., 2001; Liu et al., 2015), bulls (Gholami et al., 2010; Moallem et al., 2015; Tran et al., 2016), and rams (Samadian et al., 2010; Esmaeili et al., 2012). Additionally, the quality of roasted nuts and oilseeds is significantly influenced by the roasting conditions, which enhance the product's flavour, brown colour, texture, and overall acceptability (Pittia et al., 2001; Khan and Saini, 2016). Studies show that adding n-3 polyunsaturated fatty acids to the diet can boost sperm traits associated to male fertility. Still, there’s not much information is available on how male dairy bulls in tropical India respond to diets rich in both omega-6 and omega-3 fatty acids. Crossbred bulls, in particular, often have trouble with semen quality and freezing (Mukhopadhyay et al., 2010), which means they need better, targeted solutions. So, finding the right nutrition and management strategies to improve their semen quality is crucial. Therefore, this study set out to investigate the effect of supplementing roasted flaxseed and freshwater misting during the summer looking at how these changes affect their semen quality in Karan Fries bulls. 2. Materials and Methods The present study was conducted on eighteen Holstein Frisian crossbred (Karan Fries) bulls from May to September at the Livestock Research Centre, Livestock Research Centre, ICAR–National Dairy Research Institute, Karnal, India. The experimental site is located at 29°42′20″ N latitude and 76°58′52.5″ E longitude, at an altitude of 254.20 meters above sea level. The region experiences ambient temperatures reaching up to 45°C in summer and falling about 2°C in winter, and a diurnal temperature fluctuation of 15-20°C. The experimental bulls, aged 4 to 5 years, were housed individually in well-ventilated pens equipped facilities for seperate feeding. They were offered a Total Mixed Ration (TMR) formulated in accordance with ICAR (2013) feeding standards. The TMR comprised of maize fodder and grain, wheat bran, rice bran, deoiled mustard cake, full-fat soybean, cottonseed cake, and wheat straw. Freshly harvested maize fodder grown at the ICAR–NDRI farm was chopped and blended with concentrate mixture and wheat straw in a proportion of 60:30:10, respectively, and offered daily at 08:50 h. Clean potable water was made available ad libitum to all animals. Representative feed and fodder samples were analyzed using the methods described by the Association of Official Agricultural Chemists (AOAC, 2005), and the results are presented in Table 1. The bulls were randomly assigned to three experimental groups, each comprising six animals. Bulls in the control group (T1) received the basal diet without flaxseed supplementation, the animals in T 2 were fed basal diet supplemented with roasted flaxseed @ 1 Kg/day, while those in T 3 received basal diet supplemented with roasted flaxseed similar to T 2 with three times freshwater misting during the hotter part of the day. For the bulls in the T 3 group, a water misting system was installed beneath the roof, positioned 3.5 meters from the floor. The system utilized Foggers (NETAFIM, Fogger Cool Pro 7.5 Ltr 20 AD) to generate fine droplets of approximately 65 microns at 0.69 Kg per square meter , promoting rapid evaporation for cooling. The foggers were fabricated from engineered thermoplastic material, ensuring they were non-clogging and resistant to corrosion. Each nozzle cluster consisted of four 4-way foggers, with a flow rate of 2.0 gallons per hour. The system operated at a pressure range of 3.5 to 5 kg/sq. cm (50 to 75 psi) to produce finer mist droplets. Fan motors, each with a power rating of 1.1 horsepower and operating at 1000 rpm, were positioned 4 meters apart. For the bull's comfort, non-slip rubber mats measuring 1.22 x 1.82 meters and 5 cm thick were provided as bedding near the feeding area. Table 1. Proximate composition of feed and fodder offered to the bulls. Particulars (%) TMR Maize Green Sorghum Green Wheat straw Dry Matter 53.3 23.2 25.2 90.6 Organic Matter 94.8 93.0 89.8 90.1 Crude Protein 14.6 9.2 8.0 3.5 Ether Extract 2.6 2.2 2.2 0.9 Ash 9.7 9.3 9.1 9.9 Neutral Detergent Fibre 48.4 64.4 63.3 75.6 Acid Detergent Fibre 26.0 33.3 37.3 42.5 2.1 Assessment of Temperature–Humidity Index (THI) in Experimental Sheds The maximum and minimum temperatures, along with wet-bulb and dry-bulb readings, were obtained from the weather station at the Climate Resilient Livestock Research Centre, Dairy Cattle Physiology Division, ICAR-NDRI, Karnal (Haryana). In addition, ambient temperatures within the experimental sheds were measured using dry-bulb and wet-bulb thermometers (Zeal, Merton, London, UK) to determine the Temperature-Humidity Index (THI). The thermometers were suspended at equal heights above the bulls’ body level using threads under the shed in each group. Temperature recordings were taken twice daily, at 08:00 and 14:00 Indian Standard Time (IST). THI values for the sheds in both groups were calculated according to the formula proposed by McDowell (1972). THI = 0.72 (wet bulb temperature °C + dry bulb temperature °C) + 40.6 2.2. Assessment of Nutrients intake Dry matter intake (DMI) of experimental bulls was measured by deducting the left over daily dry matter from the quantity offered. The Crude protein and TDN intake were estimated based on the values of proximate composition. 2.3. Recording of body weight and body condition score (BCS) The bull body weights were measured in the morning, before feeding and watering, on two consecutive days each month during the feeding trial using a digital weighing balance. Body weight and body condition score (BCS) of the bulls were measured at the start and conclusion of the experimental period. Body condition scoring was recorded using a 1 to 5 score, as mentioned in the NRC (2001), where 1 indicates emaciation and 5 indicates obesity or extreme fatness. 2.4. Measurement of blood hormones and physiological responses Blood samples for hormone analysis were collected during each fortnight between 07:00 and 08:00 hours. The physiological parameters of each bull, including rectal temperature (RT), pulse rate (PR), and respiration rate (RR), were measured bi-weekly at 08:00 and 14:00 hours. Blood samples were collected fortnightly from each bull for determination of the plasma concentration of testosterone, LH and Estradiol hormones. Blood samples were drawn from the jugular vein into 9 ml vacutainers containing lithium heparin (Vacuette®, Greiner Bio-one Gmbh, Austria). They were immediately placed on ice and centrifuged for 10 min at 5000 rpm. The plasma was separated and stored in cryovials at -80 °C till the estimation of hormones. The plasma testosterone, LH and Estradiol hormones were analyzed using the ELISA Kits (ENDOCRINE TECHNOLOGIES, INC., USA). Rectal temperature (°C) was measured using a mercury-in-glass clinical thermometer (Model: LHMED Dr. Equipment-9M Clinical Oval Thermometer Alfa), by inserting 7-8 cm into the rectum, keeping the bulb in contact with the mucosa for a minimum of 2 minutes, while avoiding contact with faeces. The respiration rate was determined by observing the movement of the calf's flank, with each inward and outward motion counted as one respiration cycle. This was recorded as breaths per minute from a distance of 1 meter. Pulse rate was determined by palpation of the middle coccygeal artery at the base of the tail, with each arterial contraction–relaxation cycle counted as a single pulse. The pulse rate was recorded twice, with a 1– to 2-minute interval between counts to minimize manual error. 2.5. Evaluation of Sexual Behaviour The sexual behaviour traits of all the Karan Fries bulls were recorded during semen collection following the methods defined by Anzar et. al. (1993) and Singh et al. (2015). The following parameters (Table 2) were recorded in the semen collection yard by direct observation. Table 2. Description of sexual behaviours recorded in experimental bulls Behavior Description Reaction Time (sec) It is the interval between the exposure of the bull to the dummy and its first mount or mounting attempt, irrespective of whether the mount results in successful ejaculation. Dismounting time (sec) It is the interval between the ejaculatory thrust and the placement of the front legs back on to the ground. Total time taken to ejaculate (sec) It is the time required for a non-stimulated male to progress from initial exposure to achieving successful ejaculation when introduced to a stimulus bull. Libido score % Mating ability score % Sexual behaviour score % Calculated according to the methods described by Singh et al. (2015) 2.6. Semen collection and evaluation Semen samples from all bulls were obtained once weekly over a 22-week period extending from May to September, using the artificial vagina technique. Ejaculate volume was recorded with graduated conical tubes, while sperm concentration (×10⁶ sperm/mL) was estimated using a haemocytometer following a 1:200 dilution in 3% (w/v) sodium chloride solution. Total sperm output per ejaculate was calculated by multiplying ejaculate volume with sperm concentration. Mass motility was evaluated by placing 10 µL of undiluted semen at two points on a pre-warmed glass slide maintained at 37 °C on a stage warmer. The samples were examined under a phase-contrast microscope (Nikon Eclipse E600, Tokyo, Japan) using a 10× objective, and mass activity was graded on a 0–5 scale following the criteria described by Matharo et al. (1985). Sperm viability and morphology were evaluated using the eosin–nigrosin staining technique as described by Evans and Maxwell (1987). Smears were prepared by blending one drop (10 µL) of semen with three drops (30 µL) of the stain on a pre-warmed glass slide. The stained preparations were examined under a light microscope at 1000× magnification (Nikon Eclipse 50i, Japan), and a minimum of 200 spermatozoa were assessed across at least four randomly selected microscopic fields. Spermatozoa that excluded the eosin–nigrosin stain were classified as viable, whereas those showing partial or complete stain uptake were considered non-viable (Memon et al., 2012). Morphological evaluation was carried out on the same stained smears, and the proportion of normal spermatozoa was calculated based on observations of 200 cells. 2.7. In vitro fertility tests 2.7.1. Evaluation of functional plasma membrane integrity Functional integrity of the sperm plasma membrane was evaluated using the hypo-osmotic swelling test (HOST) following the procedure outlined by Correa and Zavos (1994). In brief, 10 µL of semen was added to 1 mL of a hypo-osmotic solution prepared by dissolving fructose (1.351 g) and trisodium citrate (0.735 g) in 100 mL of distilled water, yielding an osmolality of 150 mOsm/kg. The mixture was incubated at 37 °C for one hour. After incubation, a drop of the homogenized sample was placed on a clean, dry glass slide, covered with a coverslip, and examined microscopically. Spermatozoa exhibiting tail curling as a result of osmotic swelling were considered HOST-positive. Approximately 200 sperm cells were evaluated across multiple microscopic fields at 400× magnification using a DIC microscope, and the results were expressed as a percentage. 2.7.2. Evaluation of Acrosomal Integrity Acrosomal integrity of spermatozoa was evaluated following the staining procedure originally described by Hancock (1952). A thin smear of extended semen was prepared on a clean, dry, grease-free glass slide and allowed to air-dry at room temperature for a minimum of 10 minutes under gentle warm airflow. The smear was then fixed by immersing the slide in 10% buffered formal saline for 15 minutes, followed by thorough washing under running tap water for 15–20 minutes and subsequent drying. The fixed smear was stained by immersion in buffered Giemsa solution for 90 minutes, briefly rinsed with distilled water, and air-dried. The stained slides were examined at 1000× magnification using a light microscope with oil immersion, without a coverslip. For each sample, approximately 200 spermatozoa were evaluated across multiple microscopic fields, and acrosome integrity was expressed as a percentage of the total sperm count. 2.8. Statistical analysis The data collected during the experiment were analyzed to assess the effects of roasted flaxseed supplementation and freshwater misting applied to the body surface of Karan Fries bulls on semen quality traits. Statistical evaluation of each variable was performed using one-way analysis of variance (ANOVA), considering dietary treatment as the fixed effect. Differences were regarded as statistically significant at P < 0.05, and results are presented as mean ± standard error of the mean (SEM). The ANOVA residuals confirmed to normality assumptions. All statistical analyses were carried out using SPSS software (version 21.0) following standard statistical procedures (Snedecor and Cochran, 1994). Treatment means were compared using Duncan’s multiple range test, while environmental parameters, including the Temperature–Humidity Index (THI), were analyzed using Student’s t -test. 3. Results 3.1. Microclimate conditions During the experimental period, the mean maximum temperature ( o C) and THI of the experimental area were 38.40 and 81.40. The corresponding mean minimum temperature ( o C) and THI were 23.60 and 79.48, as depicted in Table 3. Monthly THI values recorded from each sheds differed significantly (P<0.05) in corresponding comparisons, regardless of season and time of observation (Table 4). The mean THI values of the control shed was significantly (P<0.05) higher (T 1 and T 2 ) compared to the shed of T 3 animals, where misting was provided. Table 3. Climatic conditions of the experimental location during the experimental period. Month T max ( o C) T min ( o C) THI May 38.40 23.60 79.92 June 37.70 26.60 81.40 July 33.30 26.40 79.48 August 32.40 25.50 80.80 September 33.20 24.10 80.34 Table 4. Temperature Humidity Index (THI) of experimental sheds Month Control shed Shed with misting P- Value May 80.01 ± 0.25 73.02 ± 0.52 0.00 June 81.39 ± 0.23 72.47 ± 0.39 0.00 July 81.54 ± 0.35 71.95 ± 0.39 0.00 August 81.32 ± 0.45 73.11 ± 0.54 0.00 September 81.05 ± 0.32 73.86 ± 0.39 0.00 3.2. Nutrients intake The mean values of DMI (kg/d) were 13.08 ± 0.77, 11.98 ± 0.66, and 12.23 ± 0.72 kg in T 1 , T 2 , and T 3 , respectively. Dry matter intake did not differ significantly among the groups. Also, no effect of diet was observed on total crude protein intake during the summer season in T 1 , T 2 and T 3 (1.42 ± 0.07, 1.32 ± 0.06 and 1.35 ± 0.52 kg, respectively). The overall mean value of TDNI (kg/d) was 7.57 ± 0.41 (T 1 ), 7.11 ± 0.41 (T 2 ) and 7.08 ± 0.42 (T 3 ). 3.3. Body weight and Body condition score (BCS) The mean initial body weights of the bulls in T 1 , T 2 and T 3 were 649.17 ± 40.03, 653.50 ± 4.89 and 645.17 ± 50.38 kg, respectively. The results (Table 5) revealed no significant difference among the groups at the beginning of the experiment. Furthermore, no significant differences were observed in mean body weight among the groups at the end of the experiment. The average values of initial and final body condition scores of Karan Fries dairy bulls during the experimental period have been presented in Table 5, and no significant difference in BCS among different groups was found during the experiment. 3.4. Hormonal profile The plasma concentration of testosterone and luteinizing hormone (LH) did not differ significantly among the Karan Fries bulls across groups, as depicted in Table 5. Table 5. Performance and hormonal profile of Karan Fries bulls of different groups. Parameters T 1 T 2 T 3 P -Value Initial BW (kg) 649.17 ± 16.79 653.50 ± 8.29 645.17 ± 12.72 0.96 Final BW (Kg) 656.67 ± 27.67 650.60 ± 13.73 656.58 ± 25.15 0.72 Initial BCS 3.00 ± 0.11 3.13 ± 0.85 3.08 ± 0.83 0.89 Final BCS 3.29 ± 0.12 3.15 ± 0.15 3.33 ± 0.08 0.72 Testosterone (ng/ml) 4.85 ± 0.27 4.75 ± 0.33 4.75 ± 0.15 0.81 Luteinizing hormone (mIU/ml) 9.23 ± 0.23 8.88 ± 0.37 9.08 ± 0.36 0.69 3.5. Rectal temperature (RT), Respiration rate (RR) and Pulse rate (PR) The mean morning and afternoon values of values of rectal temperature, respiration rate and pulse rate of bulls are presented in Table 6. During the morning hours, rectal temperature was marginally higher in the T 1 group compared with T 2 and T 3 . In contrast, the afternoon rectal temperature was significantly (P < 0.01) lower in bulls of the T 3 group compared to other groups. The respiration rate of bulls measured in the morning and afternoon was significantly (P < 0.01) reduced in the T 3 group compared to the T 1 and T 2 groups A similar trend was observed for pulse rate, with significantly lower values (P < 0.01) recorded in the T3 group during both observation periods. Table 6 . Physiological responses of Karan Fries bulls managed under different groups. Parameters T 1 T 2 T 3 P-Value Rectal temperature ( 0 C) Morning 38.96 a ± 0.32 38.67 ab ± 0.36 37.83 b ± 0.30 0.01 Afternoon 40.20 a ± 0.19 39.82 a ± 0.42 38.08 b ± 0.36 0.00 Respiratory rate (No./min.) Morning 25.83 ± 0.60 27.17 ± 0.70 25.00 ± 1.06 0.09 Afternoon 31.00 a ± 0.73 30.33 a ± 0.91 27.17 b ± 0.60 0.01 Pulse rate (No./min.) Morning 66.50 a ± 1.84 67.17 a ± 1.37 59.83 b ± 1.01 0.00 Afternoon 82.33 a ± 1.81 81.50 a ± 1.17 68.33 b ± 1.12 0.00 Means bearing different superscripts in a row differ letters a, b, c significantly (P < 0.05) 3.6. Sexual Behaviour The observations of the different sexual behaviours exhibited by the bulls over the experimental period are summarized in Table 7. During the summer season, bulls in the T 3 group exhibited significantly (P<0.01) lower reaction time, dismounting time and the time taken to ejaculate compared to T. However, the reaction time in T 2 was statistically similar to that of the other groups. In contrast, the libido score, mating ability score and overall sexual behaviour score were significantly (P<0.01) higher in T 3 bulls than in remaining groups. The dietary supplementation of flax seed and misting during summer enhanced libido, mating efficiency, and sexual behaviour, while reducing reaction time, dismounting time and total time to ejaculate in Karan Fries bulls. Table 7. Sexual behaviour of Karan Fries bulls managed under different groups during summer Parameter T 1 T 2 T 3 P-value Reaction Time (sec) 40.81 b ±3.54 32.19 ab ±3.88 23.61 a ±3.01 0.01 Dismounting time (sec) 5.42 b ±0.37 4.22 b ±0.25 3.39 c ±0.20 0.01 Total time taken to ejaculate (sec) 85.83 b ±9.31 61.83 ab ±7.47 50.69 a ±5.69 0.02 Libido score % 58.89 b ±1.479 64.44 b ±1.801 69.72 c ±1.227 0.00 Mating ability score % 70.83 a ±2.370 83.33 b ±2.108 83.89 b ±2.192 0.03 Sexual behaviour score % 70.28 a ±2.41 77.22 ab ±2.28 84.44 b ±2.91 0.00 Means bearing different superscripts in a row differ letters a, b, c significantly (P < 0.05) 3.7. Seminal attributes and Sperm functional attributes The values of seminal attributes of dairy bulls under different treatments have been depicted in Table 8. Bulls in the T3 group produced a greater ejaculate volume than those in the other groups. Mass activity was higher in T 2 and T 3 compared to T 1 . Sperm concentration (million/mL) was significantly greater (P < 0.01) in the T3 group compared with T2 and T1. Similarly, significantly higher values (P < 0.05) were observed for total sperm output in the T 3 bulls relative to other groups. Fresh semen from the bulls in T 3 group exhibited significantly (P<0,05) higher sperm motility (%), proportion of live spermatozoa, proportions of live spermatozoa, normal morphological forms, HOST-positive sperm, and acrosome integrity, followed by T 2 and T 1 . Additionally, the values for T 2 were significantly higher (P < 0.05) compared to those for T 1 . The mean values of total sperm abnormalities (%) were significantly (P<0.05) lower in both T 3 and T 2 groups compared with T 1 bulls. Table 8. Seminal attributes of fresh semen of Karan Fries bulls in different groups. Parameters T 1 T 2 T 3 P-Value Ejaculate Volume (ml) 4.89 ± 0.19 4.90 ± 0.25 5.03 ± 0.20 0.06 Mass motility (0–5) 2.23 ± 0.13 2.42 ± 0.06 2.43 ± 0.07 0.06 Concentration (10 6 /ml) 983.00 a ± 28.37 997.83 a ± 24.71 1044.03 b ± 33.42 0.00 Total sperm output 4709.17 a ± 25.87 4898.83 a ± 23.01 5469.42 b ± 24.99 0.03 Motility (%) 57.25 a ± 1.70 63.60 b ±1.31 69.83 c ± 1.07 0.02 Live (%) 59.75 a ± 0.12 62.75 b ± 01.26 68.08 c ± 0.92 0.00 Normal morphology (%) 66.77 a ± 1.83 67.65 b ± 1.34 75.05 c ± 1.30 0.04 HOST (%) 59.13 a ± 2.57 62.49 b ± 1.63 75.05 c ± 1.30 0.03 Acrosome integrity (%) 74.73 a ± 1.87 77.68 b ± 1.01 81.02 c ± 0.87 0.03 Head abnormality (%) 4.55 b ± 0.44 4.17 b ± 0.29 2.82 a ± 0.17 0.01 Tail abnormality (%) 4.83 b ± 0.38 4.75 b ± 0.31 3.53 a ± 0.25 0.01 Midpiece abnormality (%) 6.68 b ± 0.41 6.83 b ± 0.37 5.08 a ± 0.29 0.01 Total abnormal Sperm (%) 16.07 b ± 1.09 15.70 a ± 0.85 11.40 a ± 0.64 0.00 Means bearing different superscripts in a row differ letters a, b, c significantly at (P < 0.05) 4. Discussion The present study aimed to evaluate the influence of dietary supplementation with flaxseed and misting on semen quality traits and sexual behaviour of Crossbred bulls. Heat stress is well recognized as a major factor adversely affecting the health and productive performance of dairy animals, including buffaloes (Marai and Haeeb, 2010; Dash et al., 2016; Prasad et al., 2022). Early and accurate detection of heat stress is crucial for timely intervention, helping to reduce economic losses while enhancing animal welfare. Exposure to thermal stress elevates testicular temperature, adversely influencing seminal and biochemical characteristics and potentially predisposing bulls to infertility. Consequently, heat stress markedly reduces conception rates and fertility per insemination in males, ultimately compromising overall reproductive fitness (Bhakat et al., 2014). Appetite in the hypothalamus is impaired by the environmental Heat stress which may lead to a reduced feed intake (Kumar, 2018). The nutrient intake in all three groups was similar, and there was no significant difference in DM intake and protein intake between the groups during the summer season. During heat stress, various physiological and behavioural strategies are triggered to eliminate the surplus heat load on the animals. This also includes a reduction of feed intake, because of lowered metabolic activities (Becker et al., 2020; Tao et al., 2020; Prasad et al., 2022). Leydig cells produce testosterone, which is vital for the development and regulation of spermatogenesis (Tran et al., 2016). Elevated THI leads to increased environmental stress, resulting in disruption of hypothalamic-pituitary-gonadal axis, resulting in decreased testosterone secretion from the testes. This decrease in testosterone is often accompanied by alterations in luteinizing hormone (LH) concentrations, as the pituitary gland attempts to compensate for the lower testosterone levels. However, prolonged heat stress may cause a decline in both testosterone and LH, disrupting their normal relationship and impairing reproductive function. However, the quantity of serum testosterone and luteinizing hormone didn't differ significantly between the two groups in the current study. Though, Minton et al. (1981) found that elevated ambient temperatures result in a temporary decrease in serum LH levels after six days, although serum testosterone levels remain essentially unchanged when the bulls are exposed. Previous studies have shown that unsaturated fatty acids can stimulate steroid hormone synthesis in vivo by upregulating the expression of steroidogenic acute regulatory (StAR) protein (Wang et al., 2000; Hughes et al., 2011) and by influencing the activity of key transcription factors involved in steroidogenesis (Wathes et al., 2007). The results in the current experiment are in contrast to the results reported by Shah et al. (2016), Fair et al. (2014), and Tran et al. (2016). This might be due to the influence of breed and age of the animals. In rams, testosterone levels increase during short days, but supplementation with flaxseed oil achieves the same concentration of testosterone as in summer months (Baiomy and Mottelib, 2009). Core body temperature, pulse rate, and respiration rate play a dual role as key elements of thermoregulation and reliable indicators of physiological equilibrium. Increased respiration rate, rectal temperature, and pulse rate reflect adaptive responses aimed at maintaining thermal balance under raising ambient temperature (Lakhani et al., 2021). Jeelani et al. (2019) similarly documented a progressive increase in these vital signs with increasing THI in crossbred cattle. In the present study, bulls in the T 3 group exhibited lower rectal temperature than in the other groups. Under hot climatic conditions, ambient temperatures often exceed body temperature, which may prevent the dissipation of the excess heat to the surroundings (Prasad et al., 2022). The observed reduction in rectal temperature is consistent with the findings of Seerapu et al. (2015), who reported lower values in animals provided with fogging systems. In the current experimental setup, bulls of T 3 group were having more opportunities to dissipate extra body heat through pressure mist than those in the other groups. Fogging systems produce fine water droplets that facilitate cooling and help moderate the thermal environment (Ambulkar et al., 2011; Prasad et al., 2022). Supporting these observations, Yadav et al. (2016) and Prasad et al. (2022) reported significant reductions in body temperature of buffaloes during summer when cooled using misting or fogging systems, primarily through evaporative heat loss. The significantly (P<0.05) lower respiration rate was observed in group T 3 compared to groups T 1 and T 2 . These findings are supported by other researchers who have confirmed that panting is rapidly activated as an effective thermoregulatory mechanism, leading to rise in respiratory rate among the animals exposed to heat stress (Korde et al., 2007; Pereira et al., 2008; Yadav et al., 2019; Wankar et al., 2019). The respiration rates of cattle and buffalo are increased by high ambient temperatures (Bianca and Findlay, 1962). Ambient temperature and respiration rate exhibit a strong positive relationship, as respiration rate increases in response to rising environmental temperatures. In this situation, as it is mentioned in the previous findings (Prasad et al., 2022), a rise respiration rate may benefit forceful heat evaporation from the body to the surroundings. Bulls in the T3 group exhibited a reduced pulse rate compared with the other groups. Similar declines in pulse rate have been reported in Murrah (Seerapu et al., 2015) and Nili Ravi buffaloes (Das et al., 2013) provided with cooling interventions such as foggers and fans during the summer months. Since pulse rate reflects sympathetic nervous system activity, a lower value suggests reduced physiological stress and improved comfort in animals exposed to effective cooling measures (Sgoifo et al., 1999). Extreme heat stress leads to physical exhaustion in bulls, which can decrease their libido, resulting in longer reaction times and an increase in the total time required for successful ejaculation. This, in turn, negatively affects sperm production (Mandal et al., 2000). Similar observations to those reported in the current experiment were made by Pal et al. (2004). The present study describes a positive trend of improved libido scores in Karan Fries bulls fed flaxseed and misted compared to bulls fed only flaxseed and Control bulls' groups. However, previous reports by Panwar and Nagpaul (1989), Adwani et al. (1992), and Pal et al. (2004) are in contrast to the current findings, which might be due to differences in feeding, management, and season. During the summer, severe heat stress leads to physical exhaustion in bulls, potentially decreasing their sexual drive. This results in longer reaction times and increased duration for successful ejaculation, ultimately affecting sperm production (Mandal et al., 2000). The hot and dry summer season has been shown to negatively influence several biophysical attributes of semen in Karan Fries bulls (Bhakat et al., 2014). Mammalian spermatozoa are particularly rich in polyunsaturated fatty acids, which cannot be synthesized endogenously and therefore must be supplied through the diet (Wathes et al., 2007). The findings from the present study indicate that dietary supplementation of whole roasted flaxseed as a source of omega-3 fatty acids, along with misting under heat stress conditions, did not result in increase in the ejaculate volume. This observation is in consistent with previous observations in buffaloes (Adeel et al. 2009), in bulls (Gholami et al., 2010) and in chicken (Cerolini et al.,2006). On the other hand, dietary flaxseed inclusion did not exert a significant influence on mass activity, sperm concentration, or total sperm output per ejaculate across the experimental groups. These results align with previus findings in buffaloes (Adeel et al., 2009) and bulls (Gholami et al., 2010). The results of the present study indicate that dietary inclusion of flaxseed as source of n-3 fatty acids exerted a beneficial influence on on sperm motility, proportion of live spermatozoa, normal sperm morphology, HOST response, and acrosome integrity in fresh semen. These observations are in rationale with already reported findings in bulls fed on omega-3 fatty acids (Jadhav, 1998; Gholami et al., 2010; Argov- Argaman et al., 2013; Moallem et al., 2015; Khoshvaght et al., 2016 and Holden et al., 2017). Omega-3 supplementation and cooling reduced the percentage of sperm abnormalities in T 3 and T 2 compared to T 1 . These findings are in contract with those previously reported by Khoshvaght et al. (2016), who reported lower abnormal sperm morphology in Holstein Friesian bulls fed n-3 PUFA-rich diets during the summer season. Flaxseed supplementation plays a vital role in reducing heat stress and oxidative stress. Percentage findings are lower compared to the earlier reports by Amanda (2011), Kumar et al. ( 2015), and Moallem et al. (2015) in bulls and buffalo bulls, which may be attributed to variations in breed, age, management practices, and seasonal conditions. However, the present findings are similar to Ghlomai et al . (2011), who observed no significant improvement in feeding omega-3 fatty acids. In contrast, Byrne et al . (2017) documented higher total sperm motility and progressive motility. 5. Conclusion The present experiment found that the dietary supplementation of roasted flaxseed combined with freshwater misting during periods of summer heat stress significantly improved the physiological, behavioural, and seminal attributes of Karan Fries bulls. Although DM intake, body weight, BCS, and major reproductive hormones (testosterone and LH) remained comparable across groups, the inclusion of roasted flaxseed and mist cooling effectively reduced rectal temperature, respiration rate, and pulse rate, thereby enhancing overall thermal comfort. Improved microclimatic conditions in the mist-cooled shed were reflected in superior sexual behaviour traits, including reduced reaction time, shorter ejaculation time, and enhanced libido and mating ability scores. The combined treatment (T3) markedly improved ejaculate quality, with higher ejaculate volume, sperm concentration, total sperm output, mass motility, sperm viability, morphological normality, functional membrane integrity, and acrosome integrity as compared to the control and flaxseed-only groups. These findings highlight the synergistic benefits of omega-3–rich flaxseed and microenvironment modification in mitigating heat stress and enhancing semen quality in dairy bulls during hot tropical summers. Overall, integrating nutritional interventions with heat abatement strategies offers a practical, cost-effective approach to improving fertility potential in crossbred bulls, thereby contributing to more efficient artificial insemination programs and enhanced reproductive performance in dairy cattle. Declarations Acknowledgments The authors express their gratitude to the Director cum vice chancellor, ICAR - National Dairy Research Institute, Karnal, for providing all facilities to conduct the study. Funding This study was supported by a general grant for PhD study at ICAR – National Dairy Research Institute, Karnal (Haryana), India. Conflict of interest The authors declare that they have no conflict of interest. Ethics approval All animal studies have been approved by the Institute Animal Ethical Committee of ICAR – National Dairy Research Institute, dated: 05.11.2016, IAEC No:156/16. Consent to participate All authors have given consent for their participation in this manuscript. Consent for Publication All authors have given consent for the publication of this manuscript. Availability of data and material Data will be available on request. Authors' contributions SHA: conducted the animal experiment; KP: drafted the manuscript and analyzed the data; PS & MB: Designed and supervised the experiment; GP: helped in drafting; RS, RK & AR: helped in laboratory analysis of samples; TKM: helped in final drafting and corrections in the manuscript . References Adeel M, Ijaz A, Aleem M, Rehman H, Yousaf MS, Jabbar MA (2009) Improvement of liquid and frozen-thawed semen quality of Nili-Ravi buffalo bulls (Bubalus bubalis) through supplementation of fat. Theriogenology 71(8): 1220-1225. Ali SH, Singh P, Prasad KC, Bhakat M, Mohanty TK, Panchabai G, Sinha R (2021). Sexual Behavior and Semen Quality of Crossbred Bulls after Supplementation of Flaxseed and with Rubber Mat Floor. Ind J Vet Sci and Biotech, 17(3): 1-4. Ambulkar DR, Nikam SD, Barmase BS, Ali SZ, Jirapure SG (2011) Effect of a high-pressure fogger system on body comfort and milk yield in Murrah buffaloes during the summer. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8156401","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":589899287,"identity":"12794a80-86c2-46c0-83aa-ad738483cc61","order_by":0,"name":"Samir Humeida Ali","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Samir","middleName":"Humeida","lastName":"Ali","suffix":""},{"id":589899288,"identity":"58ed83ae-aab4-4198-be3f-1a76a3ce4ffc","order_by":1,"name":"C Kotresh Prasad","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYDACHgjF2M/MfABIS8gQr2VmO1sCSAsP8Vo2nOcxQOLjAfw9Z499+PHHTnbDYZ7Pr27UWPAwsB8+ugGfFomzfckze9uSjWce5t1mnXMM6DCetLQb+LQY8PMYM/A2MCf2AbUY57ABtUjwmBHUwvjnT31iw2GeZ8Y5/4jRwttjzMzDdjhxwmEe5se5bURokThzLplZtu248cxmNjPm3D4JHjZCfuHvyT3M+OZPtWw//+HHn3O+1cnxsx8+hlcLckSwSYBJ/MpRtTB/IKx6FIyCUTAKRiIAAJzwRQbvgbgZAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0003-1594-5421","institution":"National Dairy Research Institute","correspondingAuthor":true,"prefix":"","firstName":"C","middleName":"Kotresh","lastName":"Prasad","suffix":""},{"id":589899289,"identity":"4bc3f95d-5132-44a6-9486-86714cfaeb99","order_by":2,"name":"Pawan Singh","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Pawan","middleName":"","lastName":"Singh","suffix":""},{"id":589899290,"identity":"57072038-d2ef-4c71-8f8c-80b7a83d508e","order_by":3,"name":"Mukesh Bhakat","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Mukesh","middleName":"","lastName":"Bhakat","suffix":""},{"id":589899291,"identity":"899b10d8-346b-4497-a041-f39dff6dd41c","order_by":4,"name":"Girish Panchbhai","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Girish","middleName":"","lastName":"Panchbhai","suffix":""},{"id":589899292,"identity":"92d1084e-e0a8-4486-9d83-7b1434c16d6d","order_by":5,"name":"Ranjana Sinha","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Ranjana","middleName":"","lastName":"Sinha","suffix":""},{"id":589899293,"identity":"0f39c789-e440-4c3d-b452-6b615157b325","order_by":6,"name":"Abdul Rahim","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Abdul","middleName":"","lastName":"Rahim","suffix":""}],"badges":[],"createdAt":"2025-11-19 14:32:42","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8156401/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8156401/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102846757,"identity":"f091fe9a-4c71-4e43-9361-09f91c9c65f1","added_by":"auto","created_at":"2026-02-17 13:25:00","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1240054,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8156401/v1/fcae0791-9b06-4114-814b-9b297ff803c4.pdf"}],"financialInterests":"","formattedTitle":"Roasted Flaxseed and Hydro-cooling Synergy: Impacts on Seminal Quality of Crossbred Bulls under Heat Stress","fulltext":[{"header":"1.\tIntroduction","content":"\u003cp\u003eThe success of the crossbred program depends on the sound bull's ability to produce quality semen and effective spermatozoa, which are essential for producing high-quality germplasm through natural mating with cows or artificial insemination using liquid and/or frozen semen (Kumar et al., 2015). Reproductive efficiency of crossbred bulls is a key determinant of the success of artificial insemination programs in the cattle industry (Diskin, 2018). Several factors influence semen quality, including diet, management practices, climatic conditions, and genetic composition (Qureshi, 2004; Rehman \u003cem\u003eet al.,\u003c/em\u003e 2014). In males, the current intensification of heat stress scenarios may become a primary factor affecting bull fertility, as environmental temperature alters spermatogenesis and thus influences the reproductive process (Casu \u003cem\u003eet al.,\u003c/em\u003e 1991). Nutrition plays a central role in reproduction by modulating gamete development, ovulation, fertilization, and metabolite- and hormone-mediated reproductive processes (Wathes \u003cem\u003eet al.,\u003c/em\u003e 2007).\u003c/p\u003e\n\u003cp\u003eFlaxseed is one of the most concentrated plant-based sources of omega-3 fatty acids, particularly alpha-linolenic acid (ALA) (Gutte et al., 2015). Flaxseed oil contains approximately 58% linolenic acid and provides potent antioxidant activity that supports animal health and physiological functions (Mourvaki et al., 2010). Flaxseed (Linum usitatissimum), a member of the family Linaceae and commonly referred to as linseed, is a rich source of alpha-linolenic acid. Dietary inclusion of flaxseed has been reported to elevate tissue levels of alpha-linolenic acid and eicosapentaenoic acid, fatty acids that play an important role in the synthesis of key reproductive hormones (Petit et al., 2004). Omega-3 polyunsaturated fatty acids, particularly docosahexaenoic acid (DHA), play a critical role in maintaining sperm membrane integrity and viability, thereby enhancing semen quality and resilience to stress in cattle (Gulliver et al., 2012). Flaxseed, as a rich dietary source of n-3 fatty acids, has been widely reported to alter sperm lipid composition and improve semen quality across species, including boars (Rooke et al., 2001; Liu et al., 2015), bulls (Gholami et al., 2010; Moallem et al., 2015; Tran et al., 2016), and rams (Samadian et al., 2010; Esmaeili et al., 2012). Additionally, the quality of roasted nuts and oilseeds is significantly influenced by the roasting conditions, which enhance the product's flavour, brown colour, texture, and overall acceptability (Pittia et al., 2001; Khan and Saini, 2016).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eStudies show that adding n-3 polyunsaturated fatty acids to the diet can boost sperm traits associated to male fertility. Still, there’s not much information is available on how male dairy bulls in tropical India respond to diets rich in both omega-6 and omega-3 fatty acids. Crossbred bulls, in particular, often have trouble with semen quality and freezing (Mukhopadhyay et al., 2010), which means they need better, targeted solutions. So, finding the right nutrition and management strategies to improve their semen quality is crucial. Therefore, this study set out to investigate the effect of supplementing roasted flaxseed and freshwater misting during the summer looking at how these changes affect their semen quality in Karan Fries bulls.\u0026nbsp;\u003c/p\u003e"},{"header":" 2. Materials and Methods","content":"\u003cp\u003eThe present study was conducted on eighteen Holstein Frisian crossbred (Karan Fries) bulls from May to September at the Livestock Research Centre, Livestock Research Centre, ICAR\u0026ndash;National Dairy Research Institute, Karnal, India. The experimental site is located at 29\u0026deg;42\u0026prime;20\u0026Prime; N latitude and 76\u0026deg;58\u0026prime;52.5\u0026Prime; E longitude, at an altitude of \u003cu\u003e254.20 meters above sea\u003c/u\u003e level. The region experiences ambient temperatures reaching up to 45\u0026deg;C in summer and falling about 2\u0026deg;C in winter, and a diurnal temperature fluctuation of 15-20\u0026deg;C.\u003c/p\u003e\n\u003cp\u003eThe experimental bulls, aged 4 to 5 years, were housed individually in well-ventilated pens equipped facilities for seperate feeding. They were offered a Total Mixed Ration (TMR) formulated in accordance with ICAR (2013) feeding standards. The TMR comprised of maize fodder and grain, wheat bran, rice bran, deoiled mustard cake, full-fat soybean, cottonseed cake, and wheat straw. Freshly harvested maize fodder grown at the ICAR\u0026ndash;NDRI farm was chopped and blended with concentrate mixture and wheat straw in a proportion of 60:30:10, respectively, and offered daily at 08:50 h. Clean potable water was made available \u003cem\u003ead libitum\u003c/em\u003e to all animals. Representative feed and fodder samples were analyzed using the methods described by the Association of Official Agricultural Chemists (AOAC, 2005), and the results are presented in Table 1. The bulls were randomly assigned to three experimental groups, each comprising six animals. Bulls in the control group (T1) received the basal diet without flaxseed supplementation, the animals in T\u003csub\u003e2\u003c/sub\u003e were fed basal diet supplemented with roasted flaxseed @ 1 Kg/day, while those in T\u003csub\u003e3\u003c/sub\u003e received basal diet supplemented with roasted flaxseed similar to T\u003csub\u003e2\u0026nbsp;\u003c/sub\u003ewith three times freshwater misting during the hotter part of the day. \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFor the bulls in the T\u003csub\u003e3\u003c/sub\u003e group, a water misting system was installed beneath the roof, positioned 3.5 meters from the floor. The system utilized Foggers (NETAFIM, Fogger Cool Pro 7.5 Ltr 20 AD) to generate fine droplets of approximately \u003cu\u003e65 microns at 0.69 Kg per square meter\u003c/u\u003e, promoting rapid evaporation for cooling. The foggers were fabricated from engineered thermoplastic material, ensuring they were non-clogging and resistant to corrosion. Each nozzle cluster consisted of four 4-way foggers, with a flow rate of 2.0 gallons per hour. The system operated at a pressure range of 3.5 to 5 kg/sq. cm (50 to 75 psi) to produce finer mist droplets. Fan motors, each with a power rating of 1.1 horsepower and operating at 1000 rpm, were positioned 4 meters apart. For the bull\u0026apos;s comfort, non-slip rubber mats measuring \u003cu\u003e1.22 x 1.82 meters and 5 cm\u003c/u\u003e thick were provided as bedding near the feeding area.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 1. Proximate composition of feed and fodder offered to the bulls.\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"86%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eParticulars (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003eTMR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eMaize Green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003eSorghum Green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eWheat straw\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eDry Matter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e53.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e23.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003e25.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e90.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eOrganic Matter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e94.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e93.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003e89.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e90.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eCrude Protein\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e14.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e9.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003e8.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e3.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eEther Extract\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003e2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eAsh\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e9.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e9.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003e9.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e9.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eNeutral Detergent Fibre\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e48.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e64.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003e63.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e75.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 30px;\"\u003e\n \u003cp\u003eAcid Detergent Fibre\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 10px;\"\u003e\n \u003cp\u003e26.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e33.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 21px;\"\u003e\n \u003cp\u003e37.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e42.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.1 Assessment of Temperature\u0026ndash;Humidity Index (THI) in Experimental Sheds\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe maximum and minimum temperatures, along with wet-bulb and dry-bulb readings, were obtained from the weather station at the Climate Resilient Livestock Research Centre, Dairy Cattle Physiology Division, ICAR-NDRI, Karnal (Haryana). In addition, ambient temperatures within the experimental sheds were measured using dry-bulb and wet-bulb thermometers (Zeal, Merton, London, UK) to determine the Temperature-Humidity Index (THI). The thermometers were suspended at equal heights above the bulls\u0026rsquo; body level using threads under the shed in each group. Temperature recordings were taken twice daily, at 08:00 and 14:00 Indian Standard Time (IST). THI values for the sheds in both groups were calculated according to the formula proposed by McDowell (1972).\u003c/p\u003e\n\u003cp\u003eTHI = 0.72 (wet bulb temperature \u0026deg;C + dry bulb temperature \u0026deg;C) + 40.6\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.2.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eAssessment of Nutrients intake\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDry matter intake (DMI) of experimental bulls was measured by deducting the left over daily dry matter from the quantity offered. The Crude protein and TDN intake were estimated based on the values of proximate composition.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.3.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eRecording of body weight and body condition score (BCS)\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe bull body weights were measured in the morning, before feeding and watering, on two consecutive days each month during the feeding trial using a digital weighing balance. Body weight and body condition score (BCS) of the bulls were measured at the start and conclusion of the experimental period. Body condition scoring was recorded using a 1 to 5 score, as mentioned in the NRC (2001), where 1 indicates emaciation and 5 indicates obesity or extreme fatness.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.4.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eMeasurement of blood hormones and physiological responses\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBlood samples for hormone analysis were collected during each fortnight between 07:00 and 08:00 hours. The physiological parameters of each bull, including rectal temperature (RT), pulse rate (PR), and respiration rate (RR), were measured bi-weekly at 08:00 and 14:00 hours. Blood samples were collected fortnightly from each bull for determination of the plasma concentration of testosterone, LH and Estradiol hormones. Blood samples were drawn from the jugular vein into 9 ml vacutainers containing lithium heparin (Vacuette\u0026reg;, Greiner Bio-one Gmbh, Austria). They were immediately placed on ice and centrifuged for 10 min at 5000 rpm. The plasma was separated and stored in cryovials at -80 \u0026deg;C till the estimation of hormones. The plasma testosterone, LH and Estradiol hormones were analyzed using the ELISA Kits (ENDOCRINE TECHNOLOGIES, INC., USA).\u003c/p\u003e\n\u003cp\u003eRectal temperature (\u0026deg;C) was measured using a mercury-in-glass clinical thermometer (Model: LHMED Dr. Equipment-9M Clinical Oval Thermometer Alfa), by inserting 7-8 cm into the rectum, keeping the bulb in contact with the mucosa for a minimum of 2 minutes, while avoiding contact with faeces. The respiration rate was determined by observing the movement of the calf\u0026apos;s flank, with each inward and outward motion counted as one respiration cycle. This was recorded as breaths per minute from a distance of 1 meter. Pulse rate was determined by palpation of the middle coccygeal artery at the base of the tail, with each arterial contraction\u0026ndash;relaxation cycle counted as a single pulse. The pulse rate was recorded twice, with a 1\u0026ndash; to 2-minute interval between counts to minimize manual error.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.5.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eEvaluation of Sexual Behaviour\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe sexual behaviour traits of all the Karan Fries bulls were recorded during semen collection following the methods defined by Anzar \u003cem\u003eet. al.\u003c/em\u003e (1993) and Singh et al. (2015). The following parameters (Table 2) were recorded in the semen collection yard by direct observation.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. Description of sexual behaviours recorded in experimental bulls\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eBehavior\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003e\u003cstrong\u003eDescription\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003eReaction Time (sec)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003eIt is the interval between the exposure of the bull to the dummy and its first mount or mounting attempt, irrespective of whether the mount results in successful ejaculation.\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003eDismounting time (sec)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003eIt is the interval between the ejaculatory thrust and the placement of the front legs back on to the ground.\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003eTotal time taken to ejaculate (sec)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 65px;\"\u003e\n \u003cp\u003eIt is the time required for a non-stimulated male to progress from initial exposure to achieving successful ejaculation when introduced to a stimulus bull. \u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 34px;\"\u003e\n \u003cp\u003eLibido score %\u003c/p\u003e\n \u003cp\u003eMating ability score %\u003c/p\u003e\n \u003cp\u003eSexual behaviour score %\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 65px;\"\u003e\n \u003cp\u003eCalculated according to the methods described by Singh et al. (2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.6. \u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eSemen collection and evaluation\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSemen samples from all bulls were obtained once weekly over a 22-week period extending from May to September, using the artificial vagina technique. Ejaculate volume was recorded with graduated conical tubes, while sperm concentration (\u0026times;10⁶ sperm/mL) was estimated using a haemocytometer following a 1:200 dilution in 3% (w/v) sodium chloride solution. Total sperm output per ejaculate was calculated by multiplying ejaculate volume with sperm concentration. Mass motility was evaluated by placing 10 \u0026micro;L of undiluted semen at two points on a pre-warmed glass slide maintained at 37 \u0026deg;C on a stage warmer. The samples were examined under a phase-contrast microscope (Nikon Eclipse E600, Tokyo, Japan) using a 10\u0026times; objective, and mass activity was graded on a 0\u0026ndash;5 scale following the criteria described by Matharo et al. (1985).\u003c/p\u003e\n\u003cp\u003eSperm viability and morphology were evaluated using the eosin\u0026ndash;nigrosin staining technique as described by Evans and Maxwell (1987). Smears were prepared by blending one drop (10 \u0026micro;L) of semen with three drops (30 \u0026micro;L) of the stain on a pre-warmed glass slide. The stained preparations were examined under a light microscope at 1000\u0026times; magnification (Nikon Eclipse 50i, Japan), and a minimum of 200 spermatozoa were assessed across at least four randomly selected microscopic fields. Spermatozoa that excluded the eosin\u0026ndash;nigrosin stain were classified as viable, whereas those showing partial or complete stain uptake were considered non-viable (Memon et al., 2012). Morphological evaluation was carried out on the same stained smears, and the proportion of normal spermatozoa was calculated based on observations of 200 cells.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.7.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eIn vitro fertility tests\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.7.1. \u0026nbsp; Evaluation of\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003efunctional plasma membrane integrity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFunctional integrity of the sperm plasma membrane was evaluated using the hypo-osmotic swelling test (HOST) following the procedure outlined by Correa and Zavos (1994). In brief, 10 \u0026micro;L of semen was added to 1 mL of a hypo-osmotic solution prepared by dissolving fructose (1.351 g) and trisodium citrate (0.735 g) in 100 mL of distilled water, yielding an osmolality of 150 mOsm/kg. The mixture was incubated at 37 \u0026deg;C for one hour. After incubation, a drop of the homogenized sample was placed on a clean, dry glass slide, covered with a coverslip, and examined microscopically. Spermatozoa exhibiting tail curling as a result of osmotic swelling were considered HOST-positive. Approximately 200 sperm cells were evaluated across multiple microscopic fields at 400\u0026times; magnification using a DIC microscope, and the results were expressed as a percentage.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e2.7.2. \u0026nbsp; Evaluation of Acrosomal Integrity\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAcrosomal integrity of spermatozoa was evaluated following the staining procedure originally described by Hancock (1952). A thin smear of extended semen was prepared on a clean, dry, grease-free glass slide and allowed to air-dry at room temperature for a minimum of 10 minutes under gentle warm airflow. The smear was then fixed by immersing the slide in 10% buffered formal saline for 15 minutes, followed by thorough washing under running tap water for 15\u0026ndash;20 minutes and subsequent drying. The fixed smear was stained by immersion in buffered Giemsa solution for 90 minutes, briefly rinsed with distilled water, and air-dried. The stained slides were examined at 1000\u0026times; magnification using a light microscope with oil immersion, without a coverslip. For each sample, approximately 200 spermatozoa were evaluated across multiple microscopic fields, and acrosome integrity was expressed as a percentage of the total sperm count.\u003c/p\u003e\n\u003cp skip=\"true\"\u003e\u003cstrong\u003e\u003cem\u003e2.8. \u0026nbsp; \u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eStatistical analysis\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data collected during the experiment were analyzed to assess the effects of roasted flaxseed supplementation and freshwater misting applied to the body surface of Karan Fries bulls on semen quality traits. Statistical evaluation of each variable was performed using one-way analysis of variance (ANOVA), considering dietary treatment as the fixed effect. Differences were regarded as statistically significant at P \u0026lt; 0.05, and results are presented as mean \u0026plusmn; standard error of the mean (SEM). The ANOVA residuals confirmed to normality assumptions. All statistical analyses were carried out using SPSS software (version 21.0) following standard statistical procedures (Snedecor and Cochran, 1994). Treatment means were compared using Duncan\u0026rsquo;s multiple range test, while environmental parameters, including the Temperature\u0026ndash;Humidity Index (THI), were analyzed using Student\u0026rsquo;s \u003cem\u003et\u003c/em\u003e-test.\u003c/p\u003e"},{"header":"3.\tResults ","content":"\u003cp\u003e\u003cstrong\u003e3.1. Microclimate conditions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDuring the experimental period, the mean maximum temperature (\u003csup\u003eo\u003c/sup\u003eC) and THI of the experimental area were 38.40 and 81.40. The corresponding mean minimum temperature (\u003csup\u003eo\u003c/sup\u003eC) and THI were 23.60 and 79.48, as depicted in Table 3. Monthly THI values recorded from each sheds differed significantly (P\u0026lt;0.05) in corresponding comparisons, regardless of season and time of observation (Table 4).\u0026nbsp;The mean THI values of the control shed was significantly (P\u0026lt;0.05) higher (T\u003csub\u003e1\u003c/sub\u003e and T\u003csub\u003e2\u003c/sub\u003e)\u003csub\u003e\u0026nbsp;\u003c/sub\u003ecompared to the shed of T\u003csub\u003e3\u003c/sub\u003e animals, where misting was provided.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Climatic conditions of the experimental location during the experimental period.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"520\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eMonth\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 128px;\"\u003e\n \u003cp\u003eT\u003csub\u003emax\u0026nbsp;\u003c/sub\u003e(\u003csup\u003eo\u003c/sup\u003eC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003eT\u003csub\u003emin\u003c/sub\u003e (\u003csup\u003eo\u003c/sup\u003eC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003eTHI\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eMay\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 128px;\"\u003e\n \u003cp\u003e38.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e23.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e79.92\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eJune\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 128px;\"\u003e\n \u003cp\u003e37.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e26.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e81.40\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eJuly\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 128px;\"\u003e\n \u003cp\u003e33.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e26.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e79.48\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eAugust\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 128px;\"\u003e\n \u003cp\u003e32.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e25.50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e80.80\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 143px;\"\u003e\n \u003cp\u003eSeptember\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 128px;\"\u003e\n \u003cp\u003e33.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 142px;\"\u003e\n \u003cp\u003e24.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 106px;\"\u003e\n \u003cp\u003e80.34\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4. Temperature Humidity Index (THI) of experimental sheds\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"518\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eMonth\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 122px;\"\u003e\n \u003cp\u003eControl shed\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 227px;\"\u003e\n \u003cp\u003eShed with misting\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e\u003cem\u003eP- Value\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eMay\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 122px;\"\u003e\n \u003cp\u003e80.01 \u0026plusmn; 0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 227px;\"\u003e\n \u003cp\u003e73.02 \u0026plusmn; 0.52\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eJune\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 122px;\"\u003e\n \u003cp\u003e81.39 \u0026plusmn; 0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 227px;\"\u003e\n \u003cp\u003e72.47 \u0026plusmn; 0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eJuly\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 122px;\"\u003e\n \u003cp\u003e81.54 \u0026plusmn; 0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 227px;\"\u003e\n \u003cp\u003e71.95 \u0026plusmn; 0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eAugust\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 122px;\"\u003e\n \u003cp\u003e81.32 \u0026plusmn; 0.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 227px;\"\u003e\n \u003cp\u003e73.11 \u0026plusmn; 0.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 104px;\"\u003e\n \u003cp\u003eSeptember\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 122px;\"\u003e\n \u003cp\u003e81.05 \u0026plusmn; 0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 227px;\"\u003e\n \u003cp\u003e73.86 \u0026plusmn; 0.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 64px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3.2.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eNutrients intake\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe mean values of DMI (kg/d) were 13.08 \u0026plusmn; 0.77, 11.98 \u0026plusmn; 0.66, and 12.23 \u0026plusmn; 0.72 kg in T\u003csub\u003e1\u003c/sub\u003e, T\u003csub\u003e2\u003c/sub\u003e, and T\u003csub\u003e3\u003c/sub\u003e, respectively. Dry matter intake did not differ significantly among the groups. Also, no effect of diet was observed on total crude protein intake during the summer season in T\u003csub\u003e1\u003c/sub\u003e, T\u003csub\u003e2\u003c/sub\u003e and T\u003csub\u003e3\u003c/sub\u003e (1.42 \u0026plusmn; 0.07, 1.32 \u0026plusmn; 0.06 and 1.35 \u0026plusmn; 0.52 kg, respectively). The overall mean value of TDNI (kg/d) was 7.57 \u0026plusmn; 0.41 (T\u003csub\u003e1\u003c/sub\u003e), 7.11 \u0026plusmn; 0.41 (T\u003csub\u003e2\u003c/sub\u003e) and 7.08 \u0026plusmn; 0.42 (T\u003csub\u003e3\u003c/sub\u003e).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3.3.\u0026nbsp;Body weight and Body condition score (BCS)\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe mean initial body weights of the bulls in T\u003csub\u003e1\u003c/sub\u003e, T\u003csub\u003e2\u003c/sub\u003e and T\u003csub\u003e3\u003c/sub\u003e were 649.17 \u0026plusmn; 40.03, 653.50 \u0026plusmn; 4.89 and 645.17 \u0026plusmn; 50.38 kg, respectively. The results (Table 5) revealed no significant difference among the groups at the beginning of the experiment. Furthermore, no significant differences were observed in mean body weight among the groups at the end of the experiment. The average values of initial and final body condition scores of Karan Fries dairy bulls during the experimental period have been presented in Table 5, and no significant difference in BCS among different groups was found during the experiment.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3.4.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eHormonal profile\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe plasma concentration of testosterone and luteinizing hormone (LH) did not differ significantly among the Karan Fries bulls across groups, as depicted in Table 5.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 5. Performance and hormonal profile of Karan Fries bulls of different groups.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eParameters\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003eP -Value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eInitial BW (kg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e649.17 \u0026plusmn; 16.79\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e653.50 \u0026plusmn; 8.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e645.17 \u0026plusmn; 12.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.96\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eFinal BW (Kg)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e656.67 \u0026plusmn; 27.67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e650.60 \u0026plusmn; 13.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e656.58 \u0026plusmn; 25.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eInitial BCS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e3.00 \u0026plusmn; 0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e3.13 \u0026plusmn; 0.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e3.08 \u0026plusmn; 0.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.89\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eFinal BCS\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e3.29 \u0026plusmn; 0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e3.15 \u0026plusmn; 0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e3.33 \u0026plusmn; 0.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.72\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eTestosterone (ng/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e4.85 \u0026plusmn; 0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e4.75 \u0026plusmn; 0.33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4.75 \u0026plusmn; 0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.81\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eLuteinizing hormone (mIU/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e9.23 \u0026plusmn; 0.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e8.88 \u0026plusmn; 0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e9.08\u0026nbsp;\u0026plusmn; 0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.69\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3.5.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eRectal temperature (RT), Respiration rate (RR)\u003c/em\u003e\u003c/strong\u003e\u003cem\u003e\u0026nbsp;\u003cstrong\u003eand Pulse rate (PR)\u003c/strong\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003eThe mean morning and afternoon values of values of rectal temperature, respiration rate and pulse rate of bulls are presented in Table 6. During the morning hours, rectal temperature was marginally higher in the T\u003csub\u003e1\u003c/sub\u003e group compared with T\u003csub\u003e2\u0026nbsp;\u003c/sub\u003eand T\u003csub\u003e3\u003c/sub\u003e. In contrast, the afternoon rectal temperature was significantly (P \u0026lt; 0.01) lower in bulls of the T\u003csub\u003e3\u003c/sub\u003e group compared to other groups. The respiration rate of bulls measured in the morning and afternoon was significantly (P \u0026lt; 0.01) reduced in the T\u003csub\u003e3\u003c/sub\u003e group compared to the T\u003csub\u003e1\u0026nbsp;\u003c/sub\u003eand T\u003csub\u003e2\u0026nbsp;\u003c/sub\u003egroups\u0026nbsp;A similar trend was observed for pulse rate, with significantly lower values (P \u0026lt; 0.01) recorded in the T3 group during both observation periods.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 6\u003c/strong\u003e. \u003cstrong\u003ePhysiological responses of Karan Fries bulls managed under different\u0026nbsp;groups.\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"638\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 207px;\"\u003e\n \u003cp\u003eParameters\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 121px;\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003eP-Value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\" style=\"width: 638px;\"\u003e\n \u003cp\u003eRectal temperature (\u003csup\u003e0\u003c/sup\u003eC)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 207px;\"\u003e\n \u003cp\u003eMorning\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 121px;\"\u003e\n \u003cp\u003e38.96\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 0.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e38.67\u003csup\u003eab\u003c/sup\u003e \u0026plusmn; 0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003e37.83\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 0.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 207px;\"\u003e\n \u003cp\u003eAfternoon\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 121px;\"\u003e\n \u003cp\u003e40.20\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 0.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 120px;\"\u003e\n \u003cp\u003e39.82\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 0.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e38.08\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\" style=\"width: 638px;\"\u003e\n \u003cp\u003eRespiratory rate (No./min.)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 207px;\"\u003e\n \u003cp\u003eMorning\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 121px;\"\u003e\n \u003cp\u003e25.83 \u0026plusmn; 0.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 120px;\"\u003e\n \u003cp\u003e27.17 \u0026plusmn; 0.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e25.00 \u0026plusmn; 1.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 207px;\"\u003e\n \u003cp\u003eAfternoon\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 121px;\"\u003e\n \u003cp\u003e31.00\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 0.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 120px;\"\u003e\n \u003cp\u003e30.33\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 0.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e27.17\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 0.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\" valign=\"top\" style=\"width: 638px;\"\u003e\n \u003cp\u003ePulse rate (No./min.)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 207px;\"\u003e\n \u003cp\u003eMorning\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 121px;\"\u003e\n \u003cp\u003e66.50\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 1.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 120px;\"\u003e\n \u003cp\u003e67.17\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 1.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 118px;\"\u003e\n \u003cp\u003e59.83\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 207px;\"\u003e\n \u003cp\u003eAfternoon\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 121px;\"\u003e\n \u003cp\u003e82.33\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 1.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 120px;\"\u003e\n \u003cp\u003e81.50\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 1.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 118px;\"\u003e\n \u003cp\u003e68.33\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 1.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 72px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans bearing different superscripts in a row differ letters \u003csup\u003ea, b, c\u003c/sup\u003e significantly (P \u0026lt; 0.05)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3.6.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eSexual Behaviour\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe observations of the different sexual behaviours exhibited by the bulls over the experimental period are summarized in Table 7. During the summer season, bulls in the T\u003csub\u003e3\u0026nbsp;\u003c/sub\u003egroup exhibited significantly (P\u0026lt;0.01)\u0026nbsp;lower reaction time, dismounting time and the time taken to ejaculate compared to T. However, the reaction time in T\u003csub\u003e2\u003c/sub\u003e was statistically similar to that of the other groups. In contrast, the libido score, mating ability score and overall sexual behaviour score were significantly (P\u0026lt;0.01) higher in T\u003csub\u003e3\u003c/sub\u003e bulls than in remaining groups. The dietary supplementation of flax seed and misting during summer enhanced libido, mating efficiency, and sexual behaviour, while reducing reaction time, dismounting time and total time to ejaculate in Karan Fries bulls.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 7. Sexual behaviour of Karan Fries bulls managed under different groups during summer\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eParameter\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eReaction Time (sec)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e40.81\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;3.54\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e32.19\u003csup\u003eab\u003c/sup\u003e\u0026plusmn;3.88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e23.61\u003csup\u003ea\u003c/sup\u003e\u0026plusmn;3.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eDismounting time (sec)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e5.42\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e4.22\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e3.39\u003csup\u003ec\u003c/sup\u003e\u0026plusmn;0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eTotal time taken to ejaculate (sec)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e85.83\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;9.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e61.83\u003csup\u003eab\u003c/sup\u003e\u0026plusmn;7.47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e50.69\u003csup\u003ea\u003c/sup\u003e\u0026plusmn;5.69\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eLibido score %\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e58.89\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;1.479\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e64.44\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;1.801\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e69.72\u003csup\u003ec\u003c/sup\u003e\u0026plusmn;1.227\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eMating ability score %\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e70.83\u003csup\u003ea\u003c/sup\u003e\u0026plusmn;2.370\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e83.33\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;2.108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e83.89\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;2.192\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 32px;\"\u003e\n \u003cp\u003eSexual behaviour score %\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 18px;\"\u003e\n \u003cp\u003e70.28\u003csup\u003ea\u003c/sup\u003e\u0026plusmn;2.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 17px;\"\u003e\n \u003cp\u003e77.22\u003csup\u003eab\u003c/sup\u003e\u0026plusmn;2.28\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 19px;\"\u003e\n \u003cp\u003e84.44\u003csup\u003eb\u003c/sup\u003e\u0026plusmn;2.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 12px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans bearing different superscripts in a row differ letters \u003csup\u003ea, b, c\u003c/sup\u003e significantly (P \u0026lt; 0.05)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003e3.7.\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e\u003cem\u003eSeminal attributes and Sperm functional attributes\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe values of seminal attributes of dairy bulls under different treatments have been depicted in Table 8. Bulls in the T3 group produced a greater ejaculate volume than those in the other groups. Mass activity was higher in T\u003csub\u003e2\u0026nbsp;\u003c/sub\u003eand T\u003csub\u003e3\u003c/sub\u003e compared to T\u003csub\u003e1\u003c/sub\u003e. Sperm concentration (million/mL) was significantly greater (P \u0026lt; 0.01) in the T3 group compared with T2 and T1. Similarly, significantly higher values (P \u0026lt; 0.05) were observed for total sperm output in the T\u003csub\u003e3\u003c/sub\u003e bulls\u003csub\u003e\u0026nbsp;\u003c/sub\u003erelative to other groups.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFresh semen from the bulls in T\u003csub\u003e3\u0026nbsp;\u003c/sub\u003egroup exhibited significantly (P\u0026lt;0,05) higher sperm motility (%), proportion of live spermatozoa, proportions of live spermatozoa, normal morphological forms, HOST-positive sperm, and acrosome integrity, followed by T\u003csub\u003e2\u003c/sub\u003e and T\u003csub\u003e1\u003c/sub\u003e. Additionally, the values for T\u003csub\u003e2\u003c/sub\u003e were significantly higher (P \u0026lt; 0.05) compared to those for T\u003csub\u003e1\u003c/sub\u003e. The mean values of total sperm abnormalities (%) were significantly (P\u0026lt;0.05) lower in both T\u003csub\u003e3\u003c/sub\u003e and T\u003csub\u003e2\u0026nbsp;\u003c/sub\u003egroups compared with T\u003csub\u003e1\u003c/sub\u003e bulls.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 8. Seminal attributes of fresh semen of Karan Fries bulls in different groups.\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellspacing=\"0\" cellpadding=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eParameters\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eT\u003csub\u003e1\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eT\u003csub\u003e2\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003eT\u003csub\u003e3\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003eP-Value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eEjaculate Volume (ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4.89 \u0026plusmn; 0.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4.90\u003csup\u003e\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e5.03 \u0026plusmn; 0.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eMass motility (0\u0026ndash;5)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e2.23 \u0026plusmn; 0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e2.42 \u0026plusmn; 0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e2.43 \u0026plusmn; 0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eConcentration (10\u003csup\u003e6\u003c/sup\u003e/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e983.00\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 28.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e997.83\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 24.71\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e1044.03\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 33.42\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eTotal sperm output\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4709.17\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 25.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4898.83\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 23.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e5469.42\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 24.99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eMotility (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e57.25\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 1.70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 20px;\"\u003e\n \u003cp\u003e63.60\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn;1.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e69.83\u003csup\u003ec\u003c/sup\u003e \u0026plusmn; 1.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eLive (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e59.75\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e62.75\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 01.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e68.08\u003csup\u003ec\u003c/sup\u003e \u0026plusmn; 0.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eNormal morphology (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e66.77\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 1.83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e67.65\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 1.34\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e75.05\u003csup\u003ec\u003c/sup\u003e \u0026plusmn; 1.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eHOST (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e59.13\u003csup\u003ea\u003c/sup\u003e \u0026plusmn; 2.57\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 20px;\"\u003e\n \u003cp\u003e62.49\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 1.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e75.05\u003csup\u003ec\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 1.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eAcrosome integrity (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e74.73\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 1.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 20px;\"\u003e\n \u003cp\u003e77.68\u003csup\u003e\u0026nbsp;b\u003c/sup\u003e \u0026plusmn; 1.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e81.02\u003csup\u003ec\u003c/sup\u003e \u0026plusmn; 0.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eHead abnormality (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4.55\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4.17\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e2.82\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eTail abnormality (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4.83\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e4.75\u003csup\u003eb\u003c/sup\u003e \u0026plusmn; 0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e3.53\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eMidpiece abnormality (%)\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e6.68\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e6.83\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e5.08\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.29\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" style=\"width: 29px;\"\u003e\n \u003cp\u003eTotal abnormal Sperm (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e16.07\u003csup\u003eb\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 1.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e15.70\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 20px;\"\u003e\n \u003cp\u003e11.40\u003csup\u003ea\u0026nbsp;\u003c/sup\u003e\u0026plusmn; 0.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" style=\"width: 9px;\"\u003e\n \u003cp\u003e0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eMeans bearing different superscripts in a row differ letters \u003csup\u003ea, b, c\u003c/sup\u003e significantly at (P \u0026lt; 0.05)\u0026nbsp;\u003c/p\u003e"},{"header":"4.\tDiscussion","content":"\u003cp\u003eThe present study aimed to evaluate the influence of dietary supplementation with flaxseed and misting on semen quality traits and sexual behaviour of Crossbred bulls. Heat stress is well recognized as a major factor adversely affecting the health and productive performance of dairy animals, including buffaloes (Marai and Haeeb, 2010; Dash et al., 2016; Prasad et al., 2022). Early and accurate detection of heat stress is crucial for timely intervention, helping to reduce economic losses while enhancing animal welfare. Exposure to thermal stress elevates testicular temperature, adversely influencing seminal and biochemical characteristics and potentially predisposing bulls to infertility. Consequently, heat stress markedly reduces conception rates and fertility per insemination in males, ultimately compromising overall reproductive fitness (Bhakat et al., 2014).\u003c/p\u003e\n\u003cp\u003eAppetite in the hypothalamus is impaired by the environmental Heat stress which may lead to a reduced feed intake (Kumar, 2018). The nutrient intake in all three groups was similar, and there was no significant difference in DM intake and protein intake between the groups during the summer season. During heat stress, various physiological and behavioural strategies are triggered to eliminate the surplus heat load on the animals. This also includes a reduction of feed intake, because of lowered metabolic activities (Becker et al., 2020; Tao et al., 2020; Prasad et al., 2022).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eLeydig cells produce testosterone, which is vital for the development and regulation of spermatogenesis (Tran et al., 2016). Elevated THI leads to increased environmental stress, resulting in disruption of hypothalamic-pituitary-gonadal axis, resulting in decreased testosterone secretion from the testes. This decrease in testosterone is often accompanied by alterations in luteinizing hormone (LH) concentrations, as the pituitary gland attempts to compensate for the lower testosterone levels. However, prolonged heat stress may cause a decline in both testosterone and LH, disrupting their normal relationship and impairing reproductive function. However, the quantity of serum testosterone and luteinizing hormone didn't differ significantly between the two groups in the current study. Though, Minton et al. (1981) found that elevated ambient temperatures result in a temporary decrease in serum LH levels after six days, although serum testosterone levels remain essentially unchanged when the bulls are exposed. Previous studies have shown that unsaturated fatty acids can stimulate steroid hormone synthesis in vivo by upregulating the expression of steroidogenic acute regulatory (StAR) protein (Wang et al., 2000; Hughes et al., 2011) and by influencing the activity of key transcription factors involved in steroidogenesis (Wathes et al., 2007). The results in the current experiment are in contrast to the results reported by Shah \u003cem\u003eet al.\u0026nbsp;\u003c/em\u003e(2016), Fair \u003cem\u003eet al.\u003c/em\u003e (2014), and Tran et al. (2016). This might be due to the influence of breed and age of the animals. In rams, testosterone levels increase during short days, but supplementation with flaxseed oil achieves the same concentration of testosterone as in summer months (Baiomy and Mottelib, 2009).\u003c/p\u003e\n\u003cp\u003eCore body temperature, pulse rate, and respiration rate play a dual role as key elements of thermoregulation and reliable indicators of physiological equilibrium. Increased respiration rate, rectal temperature, and pulse rate reflect adaptive responses aimed at maintaining thermal balance under raising ambient temperature (Lakhani et al., 2021). Jeelani et al. (2019) similarly documented a progressive increase in these vital signs with increasing THI in crossbred cattle.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn the present study, bulls in the T\u003csub\u003e3\u003c/sub\u003e group exhibited lower rectal temperature than in the other groups. Under hot climatic conditions, ambient temperatures often exceed body temperature, which may prevent the dissipation of the excess heat to the surroundings (Prasad et al., 2022). The observed reduction in rectal temperature is consistent with the findings of Seerapu et al. (2015), who reported lower values in animals provided with fogging systems. In the current experimental setup, bulls of T\u003csub\u003e3\u003c/sub\u003e group were having more opportunities to dissipate extra body heat through pressure mist than those in the other groups. Fogging systems produce fine water droplets that facilitate cooling and help moderate the thermal environment (Ambulkar et al., 2011; Prasad et al., 2022). Supporting these observations, Yadav et al. (2016) and Prasad et al. (2022) reported significant reductions in body temperature of buffaloes during summer when cooled using misting or fogging systems, primarily through evaporative heat loss.\u003c/p\u003e\n\u003cp\u003eThe significantly (P\u0026lt;0.05) lower respiration rate was observed in group T\u003csub\u003e3\u003c/sub\u003e compared to groups T\u003csub\u003e1\u003c/sub\u003e and T\u003csub\u003e2\u003c/sub\u003e. These findings are supported by other researchers who have confirmed that panting is rapidly activated as an effective thermoregulatory mechanism, leading to rise in respiratory rate among the animals exposed to heat stress (Korde et al., 2007; Pereira et al., 2008; Yadav et al., 2019; Wankar et al., 2019). The respiration rates of cattle and buffalo are increased by high ambient temperatures (Bianca and Findlay, 1962). Ambient temperature and respiration rate exhibit a strong positive relationship, as respiration rate increases in response to rising environmental temperatures. In this situation, as it is mentioned in the previous findings (Prasad et al., 2022), a rise respiration rate may benefit forceful heat evaporation from the body to the surroundings.\u003c/p\u003e\n\u003cp\u003eBulls in the T3 group exhibited a reduced pulse rate compared with the other groups. Similar declines in pulse rate have been reported in Murrah (Seerapu et al., 2015) and Nili Ravi buffaloes (Das et al., 2013) provided with cooling interventions such as foggers and fans during the summer months. Since pulse rate reflects sympathetic nervous system activity, a lower value suggests reduced physiological stress and improved comfort in animals exposed to effective cooling measures (Sgoifo et al., 1999).\u003c/p\u003e\n\u003cp\u003eExtreme heat stress leads to physical exhaustion in bulls, which can decrease their libido, resulting in longer reaction times and an increase in the total time required for successful ejaculation. This, in turn, negatively affects sperm production (Mandal et al., 2000). Similar observations to those reported in the current experiment were made by Pal \u003cem\u003eet al.\u003c/em\u003e (2004). The present study describes a positive trend of improved libido scores in Karan Fries bulls fed flaxseed and misted compared to bulls fed only flaxseed and Control bulls' groups. However, previous reports by Panwar and Nagpaul (1989), Adwani \u003cem\u003eet al.\u003c/em\u003e (1992), and Pal \u003cem\u003eet al.\u003c/em\u003e (2004) are in contrast to the current findings, which might be due to differences in feeding, management, and season. During the summer, severe heat stress leads to physical exhaustion in bulls, potentially decreasing their sexual drive. This results in longer reaction times and increased duration for successful ejaculation, ultimately affecting sperm production (Mandal et al., 2000).\u003c/p\u003e\n\u003cp\u003eThe hot and dry summer season has been shown to negatively influence several biophysical attributes of semen in Karan Fries bulls (Bhakat et al., 2014). Mammalian spermatozoa are particularly rich in polyunsaturated fatty acids, which cannot be synthesized endogenously and therefore must be supplied through the diet (Wathes et al., 2007). The findings from the present study indicate that dietary supplementation of whole roasted flaxseed as a source of omega-3 fatty acids, along with misting under heat stress conditions, did not result in increase in the ejaculate volume. This observation is in consistent with previous observations in buffaloes (Adeel et al. 2009), in bulls (Gholami et al., 2010) and in chicken (Cerolini et al.,2006). On the other hand, dietary flaxseed inclusion did not exert a significant influence on mass activity, sperm concentration, or total sperm output per ejaculate across the experimental groups. These results align with previus findings in buffaloes (Adeel et al., 2009) and bulls (Gholami et al., 2010).\u003c/p\u003e\n\u003cp\u003eThe results of the present study indicate that dietary inclusion of flaxseed as source of n-3 fatty acids exerted a beneficial influence on on sperm motility, proportion of live spermatozoa, normal sperm morphology, HOST response, and acrosome integrity in fresh semen. These observations are in rationale with already reported findings in bulls fed on omega-3 fatty acids (Jadhav, 1998; Gholami et al., 2010; Argov- Argaman \u003cem\u003eet al.,\u0026nbsp;\u003c/em\u003e2013; Moallem \u003cem\u003eet al.,\u003c/em\u003e2015; Khoshvaght \u003cem\u003eet al.,\u003c/em\u003e 2016\u0026nbsp;and Holden \u003cem\u003eet al.,\u003c/em\u003e 2017). Omega-3 supplementation and cooling reduced the percentage of sperm abnormalities in T\u003csub\u003e3\u003c/sub\u003e and T\u003csub\u003e2\u003c/sub\u003e compared to T\u003csub\u003e1\u003c/sub\u003e. These findings are in contract with those previously reported by Khoshvaght et al. (2016), who reported lower abnormal sperm morphology in Holstein Friesian bulls fed n-3 PUFA-rich diets during the summer season. Flaxseed supplementation plays a vital role in reducing heat stress and oxidative stress. Percentage findings are lower compared to the earlier reports by Amanda (2011), Kumar \u003cem\u003eet al. (\u003c/em\u003e2015), and Moallem \u003cem\u003eet al.\u003c/em\u003e (2015) in bulls and buffalo bulls, which may be attributed to variations in breed, age, management practices, and seasonal conditions. However, the present findings are similar to Ghlomai \u003cem\u003eet al\u003c/em\u003e. (2011), who observed no significant improvement in feeding omega-3 fatty acids. In contrast, Byrne \u003cem\u003eet al\u003c/em\u003e. (2017) documented higher total sperm motility and progressive motility.\u0026nbsp;\u003c/p\u003e"},{"header":"5.\tConclusion ","content":"\u003cp\u003eThe present experiment found that the dietary supplementation of roasted flaxseed combined with freshwater misting during periods of summer heat stress significantly improved the physiological, behavioural, and seminal attributes of Karan Fries bulls. Although DM intake, body weight, BCS, and major reproductive hormones (testosterone and LH) remained comparable across groups, the inclusion of roasted flaxseed and mist cooling effectively reduced rectal temperature, respiration rate, and pulse rate, thereby enhancing overall thermal comfort. Improved microclimatic conditions in the mist-cooled shed were reflected in superior sexual behaviour traits, including reduced reaction time, shorter ejaculation time, and enhanced libido and mating ability scores. The combined treatment (T3) markedly improved ejaculate quality, with higher ejaculate volume, sperm concentration, total sperm output, mass motility, sperm viability, morphological normality, functional membrane integrity, and acrosome integrity as compared to the control and flaxseed-only groups. These findings highlight the synergistic benefits of omega-3\u0026ndash;rich flaxseed and microenvironment modification in mitigating heat stress and enhancing semen quality in dairy bulls during hot tropical summers. Overall, integrating nutritional interventions with heat abatement strategies offers a practical, cost-effective approach to improving fertility potential in crossbred bulls, thereby contributing to more efficient artificial insemination programs and enhanced reproductive performance in dairy cattle.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors express their gratitude to the Director cum vice chancellor, ICAR - National Dairy Research Institute, Karnal, for providing all facilities to conduct the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by a general grant for PhD study at ICAR – National Dairy Research Institute, Karnal (Haryana), India.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll animal studies have been approved by the Institute Animal Ethical Committee of ICAR – National Dairy Research Institute, dated: \u003cu\u003e05.11.2016, IAEC No:156/16.\u003c/u\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors have given consent for their participation in this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for Publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors have given consent for the publication of this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData will be available on request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors' contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSHA: conducted the animal experiment; KP: drafted the manuscript and analyzed the data; PS \u0026amp; MB: Designed and supervised the experiment; GP: helped in drafting; RS, RK \u0026amp; AR: helped in laboratory analysis of samples; \u003cu\u003eTKM: helped in final drafting and corrections in the manuscript\u003c/u\u003e.\u0026nbsp;\u003c/p\u003e"},{"header":" References","content":"\u003col\u003e\n\u003cli\u003eAdeel M, Ijaz A, Aleem M, Rehman H, Yousaf MS, Jabbar MA (2009) Improvement of liquid and frozen-thawed semen quality of Nili-Ravi buffalo bulls (Bubalus bubalis) through supplementation of fat. 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Biol Rhythm Res. doi:10/1080/09291016.2019.16127. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-biometeorology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ijbm","sideBox":"Learn more about [International Journal of Biometeorology](http://link.springer.com/journal/484)","snPcode":"484","submissionUrl":"https://www.editorialmanager.com/ijbm/default2.aspx","title":"International Journal of Biometeorology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Libido, flaxseed, misting, semen quality, Karan Fries bull","lastPublishedDoi":"10.21203/rs.3.rs-8156401/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8156401/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis study evaluated the effects of roasted flaxseed supplementation and surface water misting on semen quality and sexual behaviour of crossbred Karan Fries bulls during the summer season under tropical Indian conditions. A total of eighteen healthy Karan Fries bulls were randomly assigned to three experimental groups (n = 6 per group). Bulls in the control group (T1) received a standard ration without any supplementation. In the second group (T2), were supplemented with roasted flaxseed ( 1 kg/bull/day), while those in the third group (T3) received the same level of flaxseed supplementation along with periodic freshwater misting over the body surface. Physiological responses including body weight, rectal temperature, respiration rate, pulse rate, and circulating concentrations of testosterone and luteinizing hormone were monitored at fortnightly intervals over two consecutive periods. Sexual behaviour traits were also recorded at the same frequency. Semen was collected fortnightly and evaluated for ejaculate volume, colour, mass activity, sperm motility, concentration, viability, hypo-osmotic swelling test (HOST) response, and acrosomal integrity in fresh samples. Statistical analysis using one-way ANOVA revealed significantly higher rectal temperature, respiration rate, and pulse rate in bulls maintained under the control group, followed by those in T2 and T3 (P \u0026lt; 0.05). In contrast, bulls in T3 exhibited significantly reduced reaction time, dismounting time, and ejaculation time compared to the control group (P \u0026lt; 0.05). Measures of sexual performance, including libido score and mating ability, were significantly superior in T3 animals. Semen quality parameters such as ejaculate volume, sperm concentration, motility, livability, percentage of normal spermatozoa, HOST response, and acrosomal integrity were also highest in T3, followed by T2 and T1 (P \u0026lt; 0.05). Overall, the findings indicate that the combined application of roasted flaxseed supplementation and freshwater misting effectively alleviates heat stress, enhances sexual behaviour, and improves semen quality in Karan Fries bulls during the summer season in tropical environments.\u003c/p\u003e","manuscriptTitle":"Roasted Flaxseed and Hydro-cooling Synergy: Impacts on Seminal Quality of Crossbred Bulls under Heat Stress","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-17 13:23:08","doi":"10.21203/rs.3.rs-8156401/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2026-02-15T17:33:18+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-02-11T23:24:16+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-02-05T11:54:05+00:00","index":"","fulltext":""},{"type":"submitted","content":"International Journal of Biometeorology","date":"2026-02-04T09:11:11+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"international-journal-of-biometeorology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ijbm","sideBox":"Learn more about [International Journal of Biometeorology](http://link.springer.com/journal/484)","snPcode":"484","submissionUrl":"https://www.editorialmanager.com/ijbm/default2.aspx","title":"International Journal of Biometeorology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"f7ee2127-6f63-4165-89cb-4fc708821091","owner":[],"postedDate":"February 17th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-02-17T13:23:08+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-17 13:23:08","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8156401","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8156401","identity":"rs-8156401","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}