Bali Bulls Research Articles

The effect of increasing proportion mixed vegetable wastes in combination with corn stover as a basal diet on intake, digestion, liveweight gain and income over feed cost of Bali bulls supplemented with rice bran

The effect of increasing proportion mixed vegetable wastes in combination with corn stover as a basal diet on intake, digestion, liveweight gain and income over feed cost of Bali bulls supplemented with rice bran

Comparative developmental competence of in vitro embryos recovered from Bali cattle with normal and poor sperm motility.

Fertility is crucial for enhancing the efficiency of livestock production, as it directly impacts the reproductive rates. A comprehensive understanding of the relationship between sperm quality and embryo development is key to optimizing reproductive outcomes and improving the quality of livestock. This study analyzed the developmental competence of in vitro embryos recovered from Bali cattle with normal or poor sperm motility. Nine bulls with normal fresh semen (NFS) or poor fresh semen (PFS) motility were ejaculated for semen. Semen ejaculates, including volume, motility, and sperm concentration, were evaluated immediately after collection to measure the quality of the fresh semen. Frozen semen was evaluated using computer-assisted semen analysis (CASA) for motility, progressive sperm motility, distance curve path, distance curve linear, distance straight line, average path velocity, curvilinear velocity, linear velocity, straightness (STR), linearity of forward progression (LIN), wobble, and average lateral head displacement (ALH). Bull groups were used to determine in vitro embryo cleavage ability after fertilization of Bali cattle. Ovaries of Bali cattle were collected by slicing, and only cytoplasmic oocytes with compact cumulus cells were used in this study. The oocytes were matured, and in vitro fertilization was performed using fertilization media with a final sperm concentration of 1.5 × 106 spermatozoa/mL. After 48 h, the embryo cleavage ability of the cultured oocytes was evaluated. There were significant differences in motility values between the NFS and PFS groups; however, there were no significant differences in the volume or sperm concentration. There was a significant difference in the LIN value between the groups but no significant differences in other CASA parameters. There were no significant differences in the cleavage rate and morula between the groups, but a positive correlation was observed between the cleavage rate and the morula and between the morula and ALH. A significant negative correlation was observed between the cleavage rate and STR and between the morula and STR; no significant differences were observed for other variables. Despite variations in sperm characteristics, both normal and poor sperm motility demonstrated comparable in vitro embryonic development competence. These findings provide important insights into the fertility potential of Bali bulls, providing valuable information that can enhance selection strategies to improve the quality of livestock production.

The Libido and Semen Quality of Bali Bulls Supplemented with Micro-Nutrients of Zinc, Selenium, Vitamin A and Vitamin E

Low quality and quantity of food intake of bulls in the tropics might adversely affect libido and semen quality through their effects on spermatogenesis process. The purpose of this study was to evaluate the effect of micro-minerals (Zn and Se) and vitamins (A and E) on the libido and semen quality of Bali bulls. 2 (two) Bali bulls, aged 4-5 years old with a body weight 300-350 kg. The study was divided into 2 periods and each period was conducted for 8 weeks. At the first period (control), the experimental bulls were fed with natural grasses and concentrates (1% of body weight). Furthermore, at the second period (treatment period), they were fed diet similar to that used in the first period with an additional of a mineral mix (Zn and Se) and vitamin (A and E). Evaluation of libido and semen quality were carried out simultaneously twice a week. Libido was measured by calculating the time required for the bulls from approaching the teaser until they mounted for the first time and from approaching until they ejaculated, respectively. Semen was collected by using an artificial vagina. The volume, colour, pH, and, viscosity, as were recorded. Evaluation of semen quality in the form of the sperm concentration, motility, viability and abnormality were done by using Computer Assisted Semen Analyser (CASA). Fresh semen was diluted by Andromed®. The diluted semen was preserved at the temperature 50C for 144 hrs. Motility, viability and abnormality were observed every 12 hrs. Data were reported as mean and standard error of mean analysed by using the paired student’s t-test. Mean interval between the bulls approaching and mounting the teaser for the first time and between approaching and ejaculation after the supplementation of micro-nutrients were significantly shorter (P<0.05) in compared to those before the supplementation (2.68 vs. 6.52 and 5.09 vs. 10.06 minutes, respectively). The sperm viability after the supplementation of micro-nutrients was significantly higher in compared to that before supplementation. (82.25 vs. 87.50 %). Preservation of diluted bull semen at the temperature of 50C for 144 hrs showed that the sperm viability was significantly higher (P<0.05) and sperm abnormality was significantly lower (P<0.05) after the supplementation of micro-nutrients in compared to those before the supplementation. It can be concluded that the supplementation micro-nutrients in Bali bulls could increase libido, sperm viability and reduce the sperm abnormality.

Quality and kinematic characteristics of Bali bulls frozen semen with purified green tea extract epigallocatechin-3-gallate antioxidant addition in diluent.

The preservation of semen quality and kinematic characteristics during cryopreservation is crucial for the reproductive success and genetic management of livestock, particularly in Bali bulls. This study aimed to investigate the effect of adding purified green tea extract antioxidant Epigallocatechin-3-gallate (EGCG) in tris egg yolk diluent on the quality and kinematic characteristics of frozen semen from Bali bulls. Fresh and frozen semen samples were obtained from Bali bull and divided into four different treatment groups. P0 contained semen samples + diluent, while P1 to P3 consisted of semen samples + diluent supplemented with EGCG levels of 0.1, 0.15, and 0.2 mg/100 ml, respectively. Data were analyzed using One-way ANOVA and followed by Duncan's test if significant differences were found (p<0.05). Parameters observed included the assessment of fresh semen quality, kinematic analysis, post-thawing sperm viability, and abnormality. The results indicated that the assessment of fresh semen quality showed macroscopic and microscopic semen quality according to SNI 4869-1:2021. Kinematic analysis revealed significant differences in DSL and STR parameters between P0 and P3 (p<0.05). EGCG supplementation also caused significant differences in motility between P0 and P3 (p<0.05). Viability and spermatozoa abnormality with EGCG supplementation did not show significant differences (p>0.05). The best results for motility, kinematics, and sperm morphology variables were found in P1 as it did not exhibit a decrease in motility, kinematics, and sperm morphology. Viability did not show significant differences between P1, P2, and P3, but the best results were found in P2 as it did not exhibit a decrease in viability with mean and standard deviation (66.84 ± 7.88). Abnormality variables also did not show significant differences between P1, P2, and P3, but the best results were found in P2 as it did not exhibit a decrease in abnormality with mean and standard deviation (23.80 ± 7.36).

Sperm HSP70: may not be an age-dependent gene but is associated with field fertility in Bali bulls (Bos sondaicus).

This study aimed to analyze the characteristics of the HSP70 gene and protein in spermatozoa of Bali bulls of different age groups and to examine its potential as a biomarker determining bull fertility. This study used frozen semen produced from six Bali bulls divided into two groups based on age (≤ 9 years and ≥ 12 years). Parameters of frozen semen quality analyzed included sperm motility and kinetics using computer-assisted semen analysis, sperm morphological defects using Diff-Quick staining, acrosome integrity using FITC-PNA staining, and DNA fragmentation using acridine orange staining. HSP70 gene expression characterization was analyzed using qRT-PCR, and HSP70 protein abundance was analyzed using enzyme immunoassays. Fertility field data were obtained by analyzing the percentage conception rate for each bull based on the artificial insemination service data contained in the Indonesian-integrated system of the National Animal Health Information System (iSIKHNAS). The results showed significant differences (P<0.05) in total and progressive motility, morphological defects of the neck and midpiece, and tail of sperm, and acrosome integrity between the age groups of Bali bulls. HSP70 gene expression and protein abundance showed no significant differences (P>0.05) in different age groups. HSP70 gene expression correlated with fertility rate (P<0.05). Age affected several semen quality parameters but did not affect HSP70 gene expression and protein abundance. The HSP70 gene molecule could be a biomarker that determines the fertility of Bali bulls.

Effect of Storage Periods on DNA Fragmentation of Post-Thawed Bali Bull Sperm

Effect of Storage Periods on DNA Fragmentation of Post-Thawed Bali Bull Sperm

Integrity of Sperm Cell Bali Bull Preserved at 5°C Using a Diluent Formulation of Extracted Red Fruit and Coconut Water

This study aims to determine the role of the best diluent in protecting the integrity of the spermatozoa membrane after storage using an alternative diluent formulation of red fruit extract (Pandanus Conoideus Lamk) and coconut water. An alternative diluent formulation featuring red fruit extract (Pandanus Conoideus Lamk) and coconut water was utilized. Fresh semen from Bali Bull was evaluated in the Integrated Field Laboratory, Department of Animal Science of Universitas Muhammadiyah Gorontalo. Bali bull sperm of high quality was diluted using a combination diluent containing red fruit extract (Pandanus Conoideus Lamk), green coconut water, and CEP-2 mixed with 10% egg yolk. Technical term abbreviations were explained at their initial use. Spermatozoa membrane integrity was assessed using the hypoosmotic swelling test (HOST), while chlortetracycline (CTC) fluorescent dye was used to evaluate spermatozoa capacitation and acrosome reaction. The randomized group design (RAK) was used in the research, consisting of four conditions and 10 replications. P0 served as the control group, which received CEP-2 plus 10% egg yolk. P1 was administered EBM diluent AK enriched with 15% KT, while P2 received EBM diluent AK plus 15% KT and fructose 1 mg/ml. P3 was given EBM diluent AK plus 15% KT and fructose 2 mg/ml. Analysis of variance was used to examine the resulting data. The findings indicated that CEP-2 plus 10% egg yolk diluent was more effective than the diluent formulations containing red fruit extract and coconut water. Spermatozoa with intact membranes and no capacitation remain high, while those with capacitation and acrosome reaction remain low.

KUALITAS SEMEN BEKU SAPI BALI PASCA THAWING DALAM WAKTU BERBEDA (Quality of Bali Bull Frozen Semen Post Thawing in Differents Times)

KUALITAS SEMEN BEKU SAPI BALI PASCA THAWING DALAM WAKTU BERBEDA (Quality of Bali Bull Frozen Semen Post Thawing in Differents Times)

Sperm Kinematics and Morphology of Bali Bull (Bos javanicus) after Freezing and Thawing Treated with Green Tea Extract in Extender

Sperm Kinematics and Morphology of Bali Bull (Bos javanicus) after Freezing and Thawing Treated with Green Tea Extract in Extender

Characteristics and Kinematics of Bali Bull Sperms after Thawing Using Tris Soy Lecithin

Soy functions as an extracellular cryoprotectant, which can maintain the integrity of spermatozoa cell membranes with its main content is lecithin. Lecithin from soybeans protects sperm cells from cold stress and reduces the effects of oxidative stress during cryopreservation. This study aimed to analyse the effect of various levels of lecithin diluent on the quality of Bali bull semen during cryopreservation. Semen collection of Bali bull was carried out once a week during four times consecutively using an artificial vagina. The semen was then diluted using the essential ingredient Tris Aminomethan with the addition of powdered soy lecithin; P1 (1 %), P2 (3 %) and P3 (5 %), respectively. Andromed® (K1) as a positive control and Tris without soy lecithin (K2) as a negative control. The parameters observed were motility, progressive motility, kinematics, viability, membrane integrity, and acrosome integrity. The results of this study showed that the dilution of semen with soy lecithin before and after thawing the semen quality was not significantly different (P&lt; 0.05) in motility, viability, plasma membrane integrity, and acrosome integrity. Meanwhile, the kinematics of VAP, VCL, VSL, DAP, DCL, and DSL showed that the average quality increased at P3 compared to K1, K2, P1, and P2, which decreased after thawing or were significantly different (P&lt; 0.05). It can be concluded that Bali bull semen diluted with 3% and 5% of tris soy lecithin produces good characteristics and kinematics, can protect spermatozoa from cold shock.

Profiling of HSPB9 gene sequences in Donggala (Bos indicus) and Polled Bali (Bos javanicus) cattle breeds of Indonesia

HSPB9 gene plays important role to protect cells from heat stress in mammals. This research aim to describe the DNA sequence of HSPB9 gene in Donggala (Bos indicus) and Bali (Bos javanicus) bulls using forward sequencing method. A total of 10 blood samples from 6 Donggala and 4 Polled Bali were used in this study for DNA analysis. The bulls were kept at the breeding station at Central Sulawesi and South Sulawesi for Donggala and Bali, respectively. Results showed that a novel nonsence mutation of c.216C>T (p.L68L) was detected in the exon 1 of Bali HSPB9 gene. Commonly, the HSPB9 gene was monomorphic in Donggala and Bali cattle. In conclusion, the mutation of c.216C>T might be used as the genetic marker to discriminate Donggala and Bali cattle.

Sperm quality of Bali bull following sexing and freezing using different cryoprotectants

The objective of this experiment was to assess the impact of sperm sexing duration on sperm quality and to compare the effectiveness of glycerol (GC) and ethylene glycol (EG) as cryoprotectants to preserve the quality of sexed sperm during the freezing and storage in liquid nitrogen. The treatments consisted of three different durations of sexing: 40 min.(D-40), 50 min (D-50), 60 min. (D-60). The variable measured was the motility and viability of sperm following sexing and post-thawing motility of Bali bull sexed sperm cryopreserved in GC and EG. The results showed that sperm motility after sexing was significantly higher in the treatment of 40 and 50 min. compared to 60 min. of sexing duration. While the viability of sexed sperm in all treatments was not significantly different. Post-thawing motility of frozen sexed sperm cryopreserved with GS was higher than sexed sperm cryopreserved with EG. Finally, it was concluded that sexing times of 40 and 50 min. were the best time to get high sperm motility and viability of Bali bull compared to sexing times of 60 min. In addition, GS could maintain the motility of sexed sperm better than EG.

The Effect of Alpinia Purpurata K. Schum Rhizome Extract Supplementation in Tris-Egg Yolk Extender on the Quality of Cryopreserved Bali Bull Sperm

The Effect of Alpinia Purpurata K. Schum Rhizome Extract Supplementation in Tris-Egg Yolk Extender on the Quality of Cryopreserved Bali Bull Sperm

PENGARUH BREED TERHADAP MOTILITAS SPERMA SAPI LOKAL INDONESIA DAN Bos taurus DI BALAI INSEMINASI BUATAN (Effect breeds on sperm motility of indonesian local bulls and bos taurus at Artificial Insemination Center)

The attempt to achieved the goal could be through using artificial insemination (AI) by frozen semen. This study aims to evaluate the effect of the breed on sperm motility using Madura, Bali bulls and Bos taurus frozen semen. The experiment used local bull frozen semen (Madura and Bali bull breeds) and Bos taurus bull (Simmental and Limousin breeds) produced by Singosari’s artificial insemination center. Sperm motility evaluation was conducted using computer-assisted sperm analysis (CASA, Spermvision, Minitube®, Germany). The analyzing data uses a generalized linear model, one-way ANOVA using the SPSS 26 software. Duncan’s multiple range test (DMRT) was used to determine any significant differences. The result shows for total motility (TM) and progressive motility (PM) are 63.89±1.72% and 54.38±4.39% (Madura bull), 51.48±3.30% and 42.47±5.48% (Bali bull), 58.21±1.13% and 44.38±3.15% (Simmental bull), also 58.12±1.64% and 51.49±3.03% (Limousin bull). There is no differential sperm motility (P&gt;0.005) in both parameters for all breeds. All frozen semen used in this study qualifies with Indonesian National Standards and is approved for use in the artificial insemination program.

Reproductive Performance of Bali Cows through Artificial Insemination Program in Polewali Mandar Regency, West Sulawesi Province

Intensification of artificial insemination (AI) in Bali cattle is a government program to increase the population and productivity of local cattle. The objective of this study was to examine the reproductive performance of Bali cows through the AI program in Polewali Mandar Regency, West Sulawesi. The study was conducted in two different locations, i.e. in the low lands (&lt;300mASL) including Wonomulyo and Campalagian Districs, and high lands (&gt;300mASL) including Limboro and Bulo Districts. Data collection was carried out on farmers and inseminators profiles, and cow’s reproductive parameters including service per conception (S/C), conception rate (CR), calving rate (CvR), calving interval (CI) and Calf Mortality (CM). These data were calculated and analyzed descriptively. The cows in this study were raised by farmers in small-scale farm, mostly under 5 heads. The cattle breeding through AI program was carried out by inseminators with over 10 years of experience. In the lowlands, S/C values were 1.26-1.17 and CI were 360 to 366.31 days, respectively. Bali cows that were inseminated using Bali bull semen had a CvR of 75.69% which were higher than those using Simmental bull semen (CvR 49.69%;). While in the highlands, Bali cows that were inseminated with Bali bull semen tended to have higher S/C (1.30) and CI (356.95 days) values but had CvR of 61.00% and CR 77.00% which was also higher than that of Simmental bull semen (S/C 1,50; CI 412,86 days; CvR 50.00%; CR 66.67%). The high S/C in the highlands (1.50) is due to location constraints or the distance between the livestock and the inseminator's residence which is quite far, so it takes time to carry out AI services and is one of the factors that can cause the provision of AI services for female cows in heat is not optimal. In the highlands, most of the cattle are grazed in the grazing areas, making it difficult for breeders to detect estrus. In general, Bali cattle produced by AI in the low lands of Polewali Mandar district tended to have better reproductive performance than those in the high lands. The S/C in the low lands (1.26 and 1.17) is lower than that in the high lands (1.3 and 1.5). Keywords: AI, Bali cows, lower land, high land reproductive performance

The quality of fresh and frozen semen Bali bull assessed from different ejaculate volume

Semen volume is the amount of ejaculate released by male Bali cattle during one ejaculation, which consists of spermatozoa and seminal plasma. The purpose of this study was to determine the effect of Bali cattle ejaculation volume on the quality of fresh and frozen semen. Semen collection was carried out using an artificial vaginal collection method. The collected semen was then evaluated with microscopic parameters: motility, viability, and abnormalities. Semen came from 7 male Bali bulls. The data obtained were analyzed by one-way ANOVA. The results showed that there was no significant difference (P>0.05) between the volume of Bali cattle ejaculation on the quality of fresh semen and also post thawing based on the parameters tested. The sperm concentration of Bali bulls has average 935.75 ± 80.25 x 106. The average percentage of fresh semen motility was 70.79±1.26 while that for frozen was 47.83±1.89. The average viability of fresh spermatozoa was 80.04±3.68 while frozen 52.87±0.67. The average abnormality value of fresh spermatozoa was 8.54±1.88 while frozen was 23.83±4.22. Based on these results, it was concluded that the different volume of ejaculate of Bali cattle did not affect the quality of fresh semen of Bali cattle, both fresh and frozen based on the parameters of motility, viability, and abnormalities.

Plasma membrane integrity and acrosomal integrity of fresh and frozen Bali bull semen based on different ejaculate volume

Semen volume is one of the measuring points used to determine the quality of a bull’s semen in one ejaculate. Plasma Membrane Integrity (PMI) and Acrosomal Integrity (AI) are more detailed microscopic examinations and are used to determine the ability of spermatozoa to fertilize. The aim of this study was to evaluate the PMI and AI of fresh and frozen semen of Bali bull based on different ejaculate volumes. This research was conducted in March-April at Artificial Insemination Centre (AIC) in Pucak, South Sulawesi. The method used in this study was an experimental method with 4 treatments of ejaculate volume 1-3 ml; 3.1-5 ml; 5.1-7 ml; and 7.1-9 ml for 3 replicates each. The observed variables were PMI and AI. The results showed that the volume variation had no significant effect (P>0.05) on the PMI and AI values of fresh Bali cattle semen, as well as post-thawing frozen semen. The average percentage of PMI in fresh semen was 63.49±1.46% and post thawing was 37.66±2.82%, while the average AI percentage for fresh semen was 73.17±2.06% and post thawing was 50.92±2.64%. The conclusion of this study is the volume of ejaculate does not affect the plasma membrane integrity and acrosomal integrity of Bali cattle semen, both fresh and frozen.

Deteksi Urolithiasis pada Sapi Bali (Bos Sondaicus) yang Dipelihara Secara Semi Intensif di Desa Noelbaki Kecamatan Kupang Tengah

One of the Urolithiasis diseases in Bali cattle (Bos sondaicus) is crystal stones found in the urinary tract. These stones are formed in the kidney pelvis, ureter, bladder and urethra which can enlarge and can cause pain, bleeding or infection in the urinary tract forming crystals in the urine Urinary stones can move down along the ureter and enter the urinary bladder and when they occur deposition, the crystallized particles can increase in size so that they can cause disturbances in cattle. This study aims to identify and determine high levels of calcium oxalate in the kidneys can also affect the occurrence of crystal stones in male Bali cattle in Noelbaki Village, Central Kupang District, Kupang Regency. Macroscopic examination of urine sediment in Bali bulls, namely the examination of urine color, pH examination, and examination of Urine Specific Gravity and Microscopic Examination including Dicentrifuge and Calcium Oxalate Test Kit on 50 male Bali cattle. The results of the urine examination of male Bali cattle showed 5 positive types of crystals, 3 calcium oxalate and 2 sturvite.

Protein Identification of Seminal Plasma in Bali Bull (Bos javanicus).

The purpose of this study was to identify seminal plasma proteins in Bali bull and their potential as biomarkers of fertility. Semen was collected from 10 bulls aged 5-10 years using an artificial vagina. Fresh semen was then centrifuged (3000× g for 30 min). The supernatant was put into straws and stored in liquid nitrogen. The semen plasma protein concentration was determined using the Bradford method, and the protein was characterized using 1D-SDS-PAGE. Coomassie Brilliant Blue (CBB) was used to color the gel, and the molecular weight of the protein was determined using PM2700. A total of 94 proteins were identified in the seminal plasma of Bali bulls analyzed using LC-MS/MS (liquid chromatography-mass spectrometry). Proteins spermadhesin 1 (SPADH1), C-type natriuretic peptide (NPPC), clusterin (CLU), apoliprotein A-II (APOA2), inositol-3-phosphate synthase 1 (ISYNA1), and sulfhydryl oxidase 1 (QSOX1) were identified as important for fertility in Bos javanicus. These proteins may prove to be important biomarkers of fertility in Bali bulls. These proteins are important for reproductive function, which includes spermatozoa motility, capacitation, and acrosome reactions. This study provides new information about the protein content in seminal plasma in Bali bulls. The LC-MS/MS-based proteome approach that we applied in this study obtained 94 proteins. The identification of these seminal plasma proteins of Bali bulls and their potential as fertility biomarkers may have an impact on the success of future artificial insemination (AI).

Progressive Motility, DNA Fragmentation, Intact Plasma Membrane, and Acrosome Status of Frozen Semen Bali and Simmental Bulls

In increasing the population of bulls in Indonesia, it is necessary to utilize Artificial Insemination (AI) technology. AI reproductive technology aims to increase reproductive efficiency and use superior semen in livestock and can prevent the spread of reproductive diseases. Frozen semen is one of the factors that influence the success of AI. Thus, the quality of frozen semen must be maintained so fertility remains good. This study aimed to determine the quality of frozen semen of Bali and Simmental bulls at the Artificial Insemination Center, South Sulawesi. This study used frozen semen from two Bali and Simmental bulls. Parameters observed were progressive motility, deoxyribonucleic acid fragmentation (DNA), acrosome status, and intact plasma membrane. The progressive motility was tested using Computer-assisted sperm analysis (CASA Sperm Vision Minitube), the DNA fragmentation using acridine orange dye, the acrosome status using peanut agglutinin, and the intact plasma membrane staining using HOS solution. This study gives a result that the progressive motility, DNA fragmentation, and acrosome status of spermatozoa in frozen semen of Bali and Simmental bulls were not significantly different (P&gt;0.05). In contrast, the whole plasma membrane of spermatozoa in Bali bulls' frozen semen was significantly higher (P&lt;0.05) than in Simmental bulls. The results indicate that based on the parameters of progressive motility, DNA fragmentation, and acrosome status of spermatozoa, both bull breeds had the same quality of frozen semen. Meanwhile, according to the spermatozoa intact plasma membrane, the frozen semen of Bali bulls had better quality than that of Simmental bulls. Keywords: Artificial Insemination, frozen semen quality, Bali and Simmental bulls