Background Many countries have already banned the use of antibiotics in animal husbandry,making it extremely difficult to maintain animal health in livestock breeding.In the livestock industry,there is an urgent need ...Background Many countries have already banned the use of antibiotics in animal husbandry,making it extremely difficult to maintain animal health in livestock breeding.In the livestock industry,there is an urgent need to develop alternatives to antibiotics which will not lead to drug resistance on prolonged use.In this study,eighteen castrated bulls were randomly divided into two groups.The control group(CK)was fed the basal diet,while the antimicrobial peptide group(AP)was fed the basal diet supplemented with 8 g of antimicrobial peptides in the basal diet for the experimental period of 270 d.They were then slaughtered to measure production performance,and the ruminal contents were isolated for metagenomic and metabolome sequencing analysis.Result The results showed that antimicrobial peptides could improve the daily weight,carcass weight,and net meat weight of the experimental animals.Additionally,the rumen papillae diameter and the micropapillary density in the AP were significantly greater than those in the CK.Furthermore,the determination of digestive enzymes and fermentation parameters showed that the contents of protease,xylanase,andβ-glucoside in the AP were greater than those in the CK.However,lipase content in the CK was greater than that in the AP.Moreover,the content of acetate,propionate,butyrate,and valerate was found to be greater in AP than those in CK.The metagenomic analysis annotated 1993 differential microorganisms at the species level.The KEGG enrichment of these microorganisms revealed that the enrichment of drug resistance-related pathways was dramatically decreased in the AP,whereas the enrichment of immune-related pathways was significantly increased.There was also a significant reduction in the types of viruses in the AP.187 probiotics with significant differences were found,135 of which were higher in AP than in CK.It was also found that the antimicrobial mechanism of the antimicrobial peptides was quite specific.Seven low-abundance microorganisms(Acinetobactersp.Ac1271,Aequorivita soesokkakensis,Bacillus lacisalsi,Haloferax larsenii,Lysinibacillussp.3DF0063,Parabacteroidessp.217,Streptomycessp.So13.3)were found to regulate growth performance of the bull negatively.Metabolome analysis identified 45 differentially differential metabolites that significantly different between the CK and the AP groups.Seven upregulated metabolites(4-pyridoxic acid,Ala-Phe,3-ureidopropionate,hippuric acid,terephthalic acid,L-alanine,uridine 5-monophosphate)improve the growth performance of the experimental animals.To detect the interactions between the rumen microbiome and metabolism,we associated the rumen microbiome with the metabolome and found that negative regulation between the above 7 microorganisms and 7 metabolites.Conclusions This study shows that antimicrobial peptides can improve the growth performance of animals while resisting viruses and harmful bacteria and are expected to become healthy alternatives to antibiotics.We demonstrated a new antimicrobial peptides pharmacological model.We demonstrated low-abundance microorganisms may play a role by regulating the content of metabolites.展开更多
The objectives of the study were to examine the effects of draught work on carcass characteristics, meat yield potential, and distribution along primal cuts in Hararghe highland bulls fed on net energy requirement bas...The objectives of the study were to examine the effects of draught work on carcass characteristics, meat yield potential, and distribution along primal cuts in Hararghe highland bulls fed on net energy requirement basis. Twelve bulls were sorted into three groups of four animals each and assigned to three treatment hours: control (0), 4, or 6, using a complete randomized design (CRD). The results of the study show that there are no significant differences (p > 0.05) in slaughter weight, hot carcass weight (HCW), chilling loss, total edible and non-edible offal, fat thickness (FT), deboned meat yield (kg), meat to bone ratio, muscle distribution, and proportion of kidney, pelvic, and heart fat (KPH%) among the treatment groups. However, the hot carcass-based dressing percentage was significantly lowered (p < 0.05) in bulls that work 4 or 6 hours a day. Meat yield was strongly correlated with HCW (r = 0.74) and FT (r = 0.73). Therefore, the number of hours the bull spent on draught work did not negatively impact meat yield or quality. Consequently, the research suggests that utilizing Hararghe highland bulls for draught service, four or six hours a day is viable for dual advantages: crop cultivation and marketable carcass.展开更多
This study evaluated the effects of palm fat powder(PFP) and coated folic acid(CFA) on growth performance, ruminal fermentation, nutrient digestibility, microbial enzyme activity, microflora, hepatic lipid content and...This study evaluated the effects of palm fat powder(PFP) and coated folic acid(CFA) on growth performance, ruminal fermentation, nutrient digestibility, microbial enzyme activity, microflora, hepatic lipid content and gene expression in dairy bulls. Forty-eight Chinese Holstein bulls((362±12.4) days of age and(483±27.1) kg of body weight(BW)) were assigned to four groups in a completely randomized design with a 2×2 factorial arrangements. Supplemental PFP(0 or 30 g PFP kg-1 dietary dry matter(DM)) and CFA(0 or 120 mg FA d-1 as CFA) were mixed into the top one-third of a total mixed ration. The study included a 20-day adaptation period and followed by a 90-day collection period. The lower(P<0.01) feed conversion ratio with PFP or CFA addition resulted from the constant DM intake and the higher(P<0.05) average daily gain. The higher(P<0.05) ruminal p H, ether extract digestibility, microbial α-amylase activity, Butyrivibrio fibrisolvens copy, and expression of peroxisome-proliferator-activated receptor α(PPARα) and carnitine palmitoyl transferase-1(CPT1), and lower ruminal total volatile fatty acids(VFA) concentration, acetate to propionate ratio, neutral detergent fibre(NDF) digestibility, copies of total protozoa and Ruminococcus flavefaciens, and expression of sterol regulatory element binding protein-1(SREBP1) and acetyl-coenzyme A carboxylase α(ACACA) were observed for PFP addition. Supplementation with CFA increased(P<0.05) ruminal total VFA concentration, acetate to propionate ratio, digestibility of DM, organic matter, crude protein and NDF, activity of cellobiase, pectinase and α-amylase, copies of selected microbial except for total protozoa, as well as expression of PPARα, but decreased(P<0.05) ruminal p H, and expression of SREBP1 and ACACA. The PFP×CFA interaction(P<0.05) was observed for ammonia N, hepatic TG content, and m RNA expression of CPT1 and FAS. There had no significant difference in hepatic TG content when CFA was supplemented in the diet without PFP addition, the lower(P=0.001) hepatic TG content was observed when CFA was supplemented in the diet with PFP addition. The higher(P<0.05) m RNA expression of CPT1, and the lower(P<0.05) m RNA expression of FAS and ammonia N concentration were observed when CFA was supplemented in diet either without or with PFP addition. The results indicated that supplementation of CFA in PFP diet was more effective on increasing hepatic CPT1 expression, and decreasing ammonia N, hepatic TG content and FAS expression than in diet without PFP. Supplementation with PFP or CFA improved growth performance of dairy bulls by promoting nutrient utilization, microbial enzyme activity, microflora, and hepatic gene expression.展开更多
F-box and WD-40 domain protein 11(FBXW11)is an important component of the E3 ubiquitin-ligase enzyme that plays a key role in the ubiquitin-dependent regulation of spermatogenesis.In our previous research,the mRNA exp...F-box and WD-40 domain protein 11(FBXW11)is an important component of the E3 ubiquitin-ligase enzyme that plays a key role in the ubiquitin-dependent regulation of spermatogenesis.In our previous research,the mRNA expression of FBXW11 in bull sperm with high motility is significantly higher than that with low motility.In the present study,the protein expression levels of FBXW11 in bull testicular tissues with low-performance sperm quality groups were significantly higher than those in normal performance groups.The immunohistochemistry result demonstrated that FBXW11 protein was located in the periphery of Leydig cells and seminiferous tubules.Three splice variants of the FBXW11 gene,namely,FBXW11-tv1,FBXW11-tv2,and FBXW11-tv3,were identified in testicular tissues.The splicing patterns of the three variants are exon skipping.The transcript FBXW11-tv2 expressions were the highest in each sample.The low-performance groups displayed higher FBXW11-tv1 and FBXW11-tv2 transcript expressions than the normal performance groups.Two CpG islands were located within the 5’UTR and exon 1-2 region of the FBXW11 gene.Bisulfite sequencing PCR results demonstrated that the methylation levels of 11 methylation sites in the CpG island 2 from−99 to−43 in the normal performance groups were significantly lower than those in the low-performance groups.Pearson correlation analysis suggested that the CpG island 2 methylation level was negatively correlated with sperm motility and the transcript FBXW11-tv2 expression level.Our data revealed that alternative splicing and DNA methylation jointly regulated FBXW11 gene expression and were correlated with sperm quality traits during spermatogenesis in Holsteins.展开更多
Apple bull’s eye rot is caused by pathogenic Neofabraea species including N. malicorticis, N. perennans and N. alba. Fruits carrying this fungal quarantine disease are prohibited from entering China. The host plants ...Apple bull’s eye rot is caused by pathogenic Neofabraea species including N. malicorticis, N. perennans and N. alba. Fruits carrying this fungal quarantine disease are prohibited from entering China. The host plants for the pathogens include several Rosaceae fruits including apple and pear. Disease symptoms and pathogen morphology are often insufficient to determine the identity of the pathogen, particularly at the species level. In the current study, we analyzed the inter-species sequence variations in the β-tublin gene, and designed specific primers to allow PCR amplification of 554 bp fragments from pathogenic Neofabraea species. The PCR products were recovered and sequenced, and Blast search was conducted using the DNA sequences in the Genbank database. The results indicated precise PCR amplification of the target sequences from the host pathogen, which allowed unambiguous identification of the species.展开更多
Osteopontin (OPN) is a protein found at higher concentrations in the seminal plasma of bulls with above average fertility. Polymorphisms have been reported within the OPN gene promoter that can affect production of th...Osteopontin (OPN) is a protein found at higher concentrations in the seminal plasma of bulls with above average fertility. Polymorphisms have been reported within the OPN gene promoter that can affect production of this protein and thus, affect fertility. Therefore, Angus (n = 5) and Angus x Gelbvieh (Balancer, n = 14) and Angus x Brahman (n = 15) bulls were evaluated for presence of single nucleotide polymorphisms (SNP) in the Bos taurus OPN gene (GenBank: AY878328.1) promoter region, and their possible effects on bull semen quality as evaluated by computer-assisted semen analysis (CASA). Semen was collected by electroejaculation 6 to 9 times from each bull, and each semen collection was evaluated by CASA for motile, progressive and rapid sperm within 5 mins of ejaculation. The bulls were genotyped for reported single nucleotide polymorphisms (SNP) in the promoter region of the OPN gene through amplification of two 700 base pair (bp) DNA fragments and sequencing of the resulting PCR products. Seven SNP sites were identified, at bp 3379, 3490, 3492, 5075, 5205, 5209, and 5263 of the OPN gene. The SNP identified at bp 5205, 5209 and 5263 had not been previously reported. Individual SNP sites were evaluated as the main effect on CASA sperm motility variables in a SAS MIXED model for repeated measures. A thymine to guanine substitution at bp 3379 was associated with increased (P ≤ 0.02) percentage of motile, progressive and rapid sperm in Angus x Brahman bulls, and tended (P ≤ 0.10) to increase the same sperm motility parameters in Angus, and Angus x Gelbvieh bulls. The percentages of motile, progressive and rapid sperm were similar (P ≥ 0.05) among genotypes for the other 6 SNP identified. These results suggest that identification and genotyping of polymorphisms within the promoter region of the bovine OPN gene may be useful for selecting bulls with improved sperm motility parameters.展开更多
Background: Semen evaluation is used to estimate the testicular function. In bulls, the spermatozoa present in the ejaculate are the result of a process that begun more than 2 mo earlier, bequeathing a delayed depicti...Background: Semen evaluation is used to estimate the testicular function. In bulls, the spermatozoa present in the ejaculate are the result of a process that begun more than 2 mo earlier, bequeathing a delayed depiction of the actual function of the testis. Since testis vascularization might be critical for the gonad function, selected pulse wave Doppler ultrasound parameters were assessed in this study, for instance the peak systolic velocity, the end diastolic velocity and the resistive index of the testicular artery along the spermatic cord, the marginal portion of the testicular artery and the intratesticular branches of the testicular artery both in healthy adult and young bulls.Correlations between these parameters and characteristics of semen that was collected numerous times, before and after the Doppler ultrasound examination.Results: The peak systolic velocity and the end diastolic velocity measured in the testicular artery along the spermatic cord(supratesticular artery – SA) were variable among the bulls and within individual bulls, likely due to the convoluted course of the vessel. The resistive index was found highly repeatable in the same bull. A reduction in the resistive index was found between the supratesticular artery and the marginal portion of the testicular artery(P < 0.01), and between the marginal portion of the testicular artery and the intratesticular branches of the testicular artery(P < 0.05). No differences were recorded for the pulse wave Doppler ultrasound parameters in young bulls compared with adults. A significant correlation was found between the resistive index of the marginal portion of the testicular artery and total sperm in the ejaculate(r = 0.516, P < 0.05), the immature sperm(r = 0.462, P < 0.05), the teratoid sperm(r = 0.375, P < 0.05), and the "Dag defect" sperm(r = 0.389, P < 0.05). Similarly, the resistive index of the intratesticular branches of the testicular artery were found correlated with the total sperm number in the ejaculate(r = 0.568, P < 0.05), the immature sperm(r = 0.523, P < 0.05), the teratoid sperm(r = 0.418, P < 0.05), and the "Dag defect" sperm(r = 0.341, P < 0.05).Conclusions: The data presented in this study suggest that the resistive index, measured at the marginal portion of the testicular artery, could be an easy-to-perform parameter to evaluate the spermatogenesis quality in young bulls and normal adults.展开更多
Aimed at the disadvantages of secondary damage to oil layers caused by the conventional bull-heading water control technique, a thermo-sensitive temporary plugging agent for water control was synthesized by water solu...Aimed at the disadvantages of secondary damage to oil layers caused by the conventional bull-heading water control technique, a thermo-sensitive temporary plugging agent for water control was synthesized by water solution polymerization and applied in the field with a new secondary temporary plugging technique. The optimization and performance evaluation of thermo-sensitive temporary plugging agent were carried out through laboratory experiments. The optimized formula is as follows:(6%-8%) acrylamide +(0.08%-0.12%) ammonium persulfate +(1.5%-2.0%) sepiolite +(0.5%-0.8%) polyethylene glycol diacrylate. The thermo-sensitive temporary plugging agent is suitable for formation temperatures of 70-90 ?C, it has high temporary plugging strength(5-40 k Pa), controllable degradation time(1-15 d), the apparent viscosity after degradation of less than 100 m Pa?S and the permeability recovery value of simulated cores of more than 95%. Based on the research results, secondary temporary plugging technique was used in a horizontal well in the Jidong Oilfield. After treatment, the well saw a drop of water cut to 27%, and now it has a water cut of 67%, its daily increased oil production was 4.8 t, and the cumulative oil increment was 750 t, demonstrating that the technique worked well in controlling water production and increasing oil production.展开更多
Background:Genetic selection in cattle has been directed to increase milk production.This,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have l...Background:Genetic selection in cattle has been directed to increase milk production.This,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have led to a reduction of fertility rates over the years.Thus,seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive efficiency.In humans,sperm chromatin condensation evaluated through chromomycin A3(CMA3)has recently been purported to be a powerful biomarker for sperm functional status and male infertility.The objectives of the present study were:a)to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability,and b)to test whether this parameter could be used as a predictor of in vivo fertility in bulls.The study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein males.Reproductive outcomes of each sire were determined by non-return rates,which were used to classify bulls into two groups(highly fertile and subfertile).Results:Chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow cytometry.Sperm quality parameters(morphology,viability,total and progressive motility)were also assessed.Pearson correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo fertility.Sperm morphology,viability and total motility presented an area under the ROC curve(AUC)of 0.54,0.64 and 0.68,respectively(P>0.05),and thus were not able to discriminate between fertile and subfertile individuals.Alternatively,while the percentage of progressively motile sperm showed a significant predictive value,with an AUC of 0.73(P=0.05),CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in bulls.Specifically,the percentage of viable sperm with poor chromatin condensation showed better accuracy and precision to predict in vivo fertility,with an AUC of 0.78(P=0.02).Conclusions:Chromatin condensation evaluated through CMA3/Yo-Pro-1 and flow cytometry is defined here as a more powerful tool than conventional sperm parameters to predict bull in vivo fertility,with a potential ability to maximising the efficiency of dairy breeding industry.展开更多
[Object]This study aimed to examine the effects of 3,7-dimethyl-1-(5-oxo-hexyl)-xanthine(pentoxifylline,PF)on the motility,mitochondrial activity,acrosome integrity and fertilization rate of spermatozoa in both fresh ...[Object]This study aimed to examine the effects of 3,7-dimethyl-1-(5-oxo-hexyl)-xanthine(pentoxifylline,PF)on the motility,mitochondrial activity,acrosome integrity and fertilization rate of spermatozoa in both fresh and frozen-thawed bull semen.[Method]Fresh and frozen-thawed bull spermatozoa were exposed to 5 mmol/L PF with untreated samples as controls.[Result]Fresh spermatozoa showed reduced(P<0.05)motility after 2 h incubation with PF whereas,surprisingly,frozen-thawed samples exhibited increased sperm motility(P<0.05)after 2 h incubation with PF and they also showed enhanced longevity compared to controls.Mitochondrial activity in both fresh and frozen-thawed bull spermatozoa increased(P<0.05)during 4 h incubation with PF whereas acrosome integrity remained unchanged in both types of semen.However,treatment with 5 mmol/L PF did not influence the in vitro fertilization efficiency of fresh spermatozoa but improved significantly(P<0.05)that of frozen-thawed spermatozoa.[Conclusion]These results indicate that PF can improve sperm quality of frozenthawed bull semen,and may improve pregnancy rates in bovine artificial insemination programmes employing frozen semen.展开更多
Objective:To evaluate the effect of adding different values of polymyxin B (PMB) to bull semen on various motility parameters of post-thawed semen such as total motility, progressive motility and velocity parameters u...Objective:To evaluate the effect of adding different values of polymyxin B (PMB) to bull semen on various motility parameters of post-thawed semen such as total motility, progressive motility and velocity parameters using kinetic parameters of sperm by Computer Assisted Sperm Analysis.Methods: Gram negative bacteria release lipopolysaccharide, which induces the apoptotic pathway. Antibiotics are added to semen in order to prevent bacterial contaminations in bovine semen. These antibiotics kill the bacteria especially gram negative bacteria. Therefore, their endotoxins are released during bacteriolysis and bind to the head region and midpiece of sperm. PMB is a bactericidal antibiotic against multidrug resistant gram-negative bacteria and is able to neutralize the toxic effects of the released endotoxin. This study was performed on 3-year old Taleshi bulls. Results:The results showed both positive and negative significant effects of PMB on semen quality. Total motility and progressive motility were significantly increased (P<0.0001) by 100 μg per mL of PMB (55.2% and 48.8% respectively) against the control groups (43.5% and 37.7%, respectively). Moreover, they were significantly decreased (P<0.0001) by 1000 μg per mL of PMB (35.2% and 28.8% respectively) against the control groups (43.5% and 37.7% respectively) in above-mentioned parameters. In Computer Assisted Semen Analyzer, parameter VAP was significantly decreased (P<0.04) in 1000 μg (69.6 μm/s) against the control group (78.7 μm/s). Finally, using PMB in processing cryopreserved bull semen is advised, but before using it, the rate of endotoxins must be measured.Conclusions: We advise using PMB after measuring endotoxin concentration;In vitro,in vivoand in field fertilization, adding other sperm evaluation factors such as acrosomal integrity, DNA integrity, mitochondrial function to PMB treated semen.展开更多
The objective of the study was to determine the level of Bruising and Dark Firm Dry on Arsi, Boran and Harar cattle breeds in Ethiopia. A total of 39 beef bulls from three different breeds (14 Boran, 14 Harar, and 11 ...The objective of the study was to determine the level of Bruising and Dark Firm Dry on Arsi, Boran and Harar cattle breeds in Ethiopia. A total of 39 beef bulls from three different breeds (14 Boran, 14 Harar, and 11 Arsi cattle breeds) were used in this study. Based on the present finding, from 39 slaughtered beef bulls 66.7% was bruised and 30.77% produced DFD meat. Harar breed had the highest (39.1%) overall level of bruising while Boran and Arsi had the lowest level. However, Harar breed had the lowest level of DFD (14.29%). Age groups 3 - 6 and 6 - 9 years had the highest level of a bruise (29.70% and 28.10%, respectively) compared to age groups 2 - 3 years. The mean number of bruises per carcass was 1.96 ± 0.25 while the bruise score per carcass was 3.58 ± 0.68. A linear bruise was the most observed (33.33%) shape of a bruise. It was concluded that the level of bruising and DFD was higher compared to some other abattoir studies in other countries which indicated the possible presence of poor handling practice of cattle from farm to abattoir gates. To minimize the incidence of DFD and bruising, it was recommended that proper handling of cattle during loading, transport, unloading, at lairriage and during slaughter need to be practiced.展开更多
Data consisting of test performance records of 725 bulls was analysed to evaluate trends for growth and feed efficiency, and to determine environmental factors that influence growth of range performance tested bulls i...Data consisting of test performance records of 725 bulls was analysed to evaluate trends for growth and feed efficiency, and to determine environmental factors that influence growth of range performance tested bulls in the arid sweet bushveld of South Africa. Performance data were subjected to regression procedures to estimate parameters of the slope (β) and intercept (α) for average daily gain (ADG) and Kleiber ratio (KR). Mixed procedures were applied to test for sources of variation in ADG and KR. There was an increase of 3.481 g/day/year and 0.528 g/day/ year in ADG for Nguni and Bonsmara bulls, respectively. The ADG trends’ respective feed efficiency as reflected by KR was 0.093 and 0.059. Herd of origin and test-year were the sources of variation for ADG while the variation in KR was due to the herd of origin, test-year, start-age and start-weight. Results indicated a steady increase in ADG and KR for range performance tested bulls in the arid sweet bushveld.展开更多
The aim of this study was to assess sperm quality (motility, viability and acrosomal integrity) sperm from commercially frozen semen straws two breeds of bulls Bos taurus (holstein Frisian) and Bos indicus (Brahman). ...The aim of this study was to assess sperm quality (motility, viability and acrosomal integrity) sperm from commercially frozen semen straws two breeds of bulls Bos taurus (holstein Frisian) and Bos indicus (Brahman). 9 commercial straws 0.5 ml of Holstein bull semen and 9 Brahman bull were thawed, they were kept for two hours at room temperature and motility, viability and acrosomal integrity (NAR) was assessed. The results were 30% motility, viability 40% and 30% of NAR in the Holstein breed. Brahma race for motility 40%, 50% and 40% viability was obtained NAR. In conclusion, according to the results of the variables analyzed, the Brahman breed in sperm quality was better than the Holstein breed;however, the results of both races meet minimum standards of quality sperm for use in artificial insemination (AI) field level.展开更多
The study was planned with an objective to assess the level of antisperm antibodies (ASA) in the blood serum and seminal plasma of breeding cow bulls and their relationship with sperm function and fertility tests. ASA...The study was planned with an objective to assess the level of antisperm antibodies (ASA) in the blood serum and seminal plasma of breeding cow bulls and their relationship with sperm function and fertility tests. ASA was analyzed in blood serum and seminal plasma by SpermMar test, Immuno peroxidase assay (IPA) and Enzyme linked immunoabsorbant assay (ELISA). In SpermMar test, about 54% bulls were with >40% IgG in blood serum against sperm surface antigens, whereas none of the bulls were with >10% IgG in seminal plasma. More than 20% and >10% IgA against sperm surface antigens were detected in the blood serum and seminal plasma of 65.8% and 37% bulls, respectively. Out of 26 bulls, seminal plasma of 21 bulls reacted with spermatozoa both in IPA and IgA latex particles and that of only 12 bulls reacted with IgG. In IPA, about 50% of the bulls had >40% ASA against head surface antigens, whereas, there were 23% bulls with >10% ASA in seminal plasma. Also ELISA indicated a higher antibody titre in blood serum (3200 - 6400) and seminal plasma (40 - 80) of 50% and 42% bulls, respectively. There were 11 bulls with low values of HOST/in vitro acrosome reaction/cervical mucus penetration assay and higher level of either serum or seminal plasma ASA. Our study revealed that a significant level of ASA in serum or seminal plasma may have effect on the fertility of bulls by affecting the sperm function.展开更多
Seminal plasma is composed of secretions from accessory sex glands, which are mixed with sperm at ejaculation and contribute the majority of semen volume. Seminal plasma is considered a transport and support medium fo...Seminal plasma is composed of secretions from accessory sex glands, which are mixed with sperm at ejaculation and contribute the majority of semen volume. Seminal plasma is considered a transport and support medium for sperm in the female reproductive tract. Because seminal plasma is not required for fertilization, the importance of its constituents to the establishment of normal pregnancy has been overlooked. Four seminal plasma proteins, Osteopontin, Sper-madhesin Z13, BSP 30 kDa and Phospholipase A2, have been identified as markers of fertility in dairy bulls (Cancel et al., 1997;Moura et al., 2006, 2007). The objective of the present study was to characterize the expression patterns of these proteins and other proteins found to be of interest in seminal plasma of cryopreserved and non-cryopreserved bull semen. Seminal plasma samples were obtained from 16 mature Hol-stein-Friesian bulls at Select Sires Inc. Samples were divided into two groups based on assigned fertility score expressed as the percentage point deviation (PD) of the bull’s non-return rate (NRR) from the average NRR of all bulls in the Select Sires Inc. reproductive management program. Group 1 (high fertility bulls, n = 8) 1.9 ≤ PD ≤ 2.7%, and group 2 (low fertility bulls, n = 8) -6.5 ≤ PD ≤ 1.8 %. Additionally, the samples were categorized as processed (cryopreserved) or unprocessed (non-cryopreserved) for protein analysis. Protein expression was analyzed by 2-D fluorescence difference gel electrophoresis (2D-DIGETM). Protein spots were picked from a reference gel, analyzed by mass spectrometry and, subsequently identified by MS/MS ion searches performed on the SwissProt database. Protein expression did not differ (P > 0.05) with fertility grouping but displayed two distinct patterns among the processing groups: majority of the functional proteins were highly expressed in seminal plasma of non-cryopreserved semen while the cryopreserved semen contained mainly structural/extender derived proteins. Functional proteins identified included Spermadhesin Z13, BSP A1/2, BSP 30 kDa, Nucleobindin-1 and metalloproteinase inhibitor 2. Some of these proteins have been identified as anti-fertility or fertility enhancing agents in males. Whether this alteration in protein expression after processing might affect semen fertility is worthy of further evaluation.展开更多
Objective:To optimize the cryopreservation of buffalo bull semen by altering freezing rates within critical temperature range (4℃ to -60℃). Methods: A total of 20 ejaculates each from 5 Murrah buffalo bulls were cry...Objective:To optimize the cryopreservation of buffalo bull semen by altering freezing rates within critical temperature range (4℃ to -60℃). Methods: A total of 20 ejaculates each from 5 Murrah buffalo bulls were cryopreserved using programmable biofreezer in 2 phases. In the 1st phase, 9 freezing rates were applied at -2, -5, -10, -20, -30, -40, -50, -60 or -4℃/min (control) from 4℃ to -15℃;at -40℃/min from -15℃ to -60℃. In the 2nd phase, a fixed freezing rate was applied at -30℃/min from 4℃ to -15℃. Six freezing rates were applied at -10, -20, -30, -40 (control), -50 or -60℃/min from -15℃ to -60℃. The freezing from -60℃ to -140℃ were fixed at -50℃/min in both the phases. Post thaw semen quality was assessed in terms of motility, viability, membrane integrity (hypo-osmotic swelling test), sperm abnormalities, and active mitochondria. Data were arc sine transformed and analyzed through one-way analysis of variance using SPSS software. Results: In the 1st phase, percent individual motility, progressive motility and viability were similar among various protocols. Percent hypo-osmotic swelling reactive sperm was higher with freezing at -30℃/min. In the 2nd phase, percent individual motility, viability and hypo-osmotic swelling reactive sperm was higher with freezing at -50℃/min. Sperm head abnormalities were lower at -30℃/min in the 1st phase, but were similar among the protocols of the 2nd phase. Percent active mitochondria were higher at -30℃/min in the 1st phase and at -50℃/min in the 2nd phase.Conclusions:The optimum post thaw semen quality of buffalo bull could be obtained by applying freezing rate at -30℃/min (4℃ to -15℃) and at -50℃/min (-15℃ to -140℃), followed by plunging of straws in into liquid nitrogen for storage.展开更多
Objective:To evaluate the supplementation effects of vitamin E,vitamin C,superoxide dismutase(SOD),and catalase to diluents on bull cryopreserved epididymal sperm.Methods:Sperm were retrieved from 20 bull testes and w...Objective:To evaluate the supplementation effects of vitamin E,vitamin C,superoxide dismutase(SOD),and catalase to diluents on bull cryopreserved epididymal sperm.Methods:Sperm were retrieved from 20 bull testes and were then supplemented with 0.1 mM vitamin E,5.0 mM vitamin C,100.0 IU/mL SOD,and 100.0μg/mL catalase alone,or in a combination.The control treatment contained no addition.After supplementation,samples were frozen and stored in liquid nitrogen.The sperm parameters including motility,progressive motility,viability,acrosome integrity,plasma membrane integrity,kinematics and DNA damage were evaluated following the thawing process.Results:Vitamin E alone significantly increased the parameters of acrosome and membrane integrity compared to the control treatment(P<0.05).While compared to the control treatment,vitamin C had no improvement effect on sperm characteristics except for membrane integrity.Treatment of vitamin E+vitamin C had a significant improvement in total motility,progressive motility,viability,membrane and acrosome integrity compared to the control and other treatments(P<0.05).Compared to the control treatment,addition of SOD or catalase alone significantly improved the percentages of total motility,progressive motility,viability,membrane and acrosome integrity(P<0.05).Furthermore,SOD+catalase significantly increased total motility,progressive motility,viability,acrosome and membrane integrity characteristics compared to the catalase treatment(P<0.05).Vitamin E alone,vitamin E+vitamin C,and SOD in diluents decreased DNA damages and thereby improved the rate of intact sperm heads.Conclusions:Addition of 100.0 IU/mL SOD alone and 0.1 mM vitamin E+5.0 mM vitamin C,and also 5.0 mM vitamin C+100μg/mL catalase in a combination improves the quality of cryopreserved bull epididymal sperm and could be used for cryopreservation.展开更多
Objective:To assess the hemodynamic changes that occurred in the main testicular artery at two different locations such as supra and marginal ones in bulls at three different ages.Methods:Eighteen Baladi bulls were di...Objective:To assess the hemodynamic changes that occurred in the main testicular artery at two different locations such as supra and marginal ones in bulls at three different ages.Methods:Eighteen Baladi bulls were divided according to their age into three different age groups:1 year[n=6;groupⅠwith bodyweight(380±10)kg],3 years[n=6;groupⅡwith bodyweight(570±10)kg],and 6 years old[n=6;groupⅢwith bodyweight(650±10)kg]Baladi bulls.Circumference of the scrotum,thickness of the mediastinum,testicular dimensions,vascularity,and hormonal levels were measured.Testicular hemodynamics were assessed by Doppler ultrasound scanning.Results:Testicular width,length,and volume were significantly different among the three age groups,with the highest in bulls of groupⅢ(P<0.05).The bulls in groupⅢhad significantly higher mediastinal line thickness and scrotal circumference than groupⅠ(P<0.05),but there was no significant difference between groupⅡand groupⅢ.Bulls in groupⅡshowed a decline in Doppler indices(resistance and pulsatility indexes)at both testicular branches compared to the other two groups(P<0.05).Supra and marginal end diastolic velocities in the three age groups were significantly different,with the highest in groupⅡ(P<0.05).Time average velocity(cm/s)was significantly elevated in groupⅡcompared to the other two groups(P<0.05).Both testicular echotextures were significantly decreased in groupⅡ(P<0.05).Estradiol and nitric oxide metabolites were significantly high in groupⅡcompared to the other two groups(P<0.05).Meanwhile,the level of plasma testosterone was maximum in groupⅢ(P<0.05).Positive correlations were found between supratesticular artery resistance and pulsatility indexes(r=0.81;P<0.001),while both resistance and pulsatility indexes had correlated negative with estradiol(r=-0.71 and r=-0.91;P<0.001),and nitric oxide metabolites(r=-0.92 and r=-0.72;P<0.001).Conclusions:Three-year old Baladi bull has the lesser Doppler indices with lesser echotexture and greater nitric oxide with estradiol concentrations,which directs a significant ability for the thermoregulation process.These parameters will help in breeding selection.展开更多
BACKGROUND Hepatocellular adenomas are rare tumors that can occur in patients with glycogen storage disease type I.CASE SUMMARY We herein report two cases of histologically proven hepatocellular adenomas in patients w...BACKGROUND Hepatocellular adenomas are rare tumors that can occur in patients with glycogen storage disease type I.CASE SUMMARY We herein report two cases of histologically proven hepatocellular adenomas in patients with glycogen storage disease type I.Magnetic resonance imaging(MRI)was performed after bolus injection of gadoxetate disodium,a liver-specific gadolinium-based MRI contrast agent.In the present cases,some of the hepatocellular adenomas showed unexpectedly a“bull’s eye”appearance on T2-weighted and post-contrast images,which was not previously described as imaging findings of hepatocellular adenomas in glycogen storage disease.A bull’s eye appearance on T2-weighted images can be encountered in both benign(i.e.,abscess)or malignant(i.e.,epithelioid hemangioendothelioma,cholangiocarcinoma,and metastases)hepatic lesions.CONCLUSION We present two cases of hepatocellular adenomas in patients with glycogen storage disease type 1,in which gadoxetate disodium-MRI showed atypical imaging findings for hepatocellular adenomas.At present there is no systematic study evaluating MRI findings of hepatocellular adenomas in patients with glycogen storage disease,further studies are needed to specifically investigate this issue.展开更多
基金financially supported by Research and application of corn straw forage and beef cattle high-efficiency and quality production technology (Provincial Education Science and Technology Innovation Project) (GSSYLXM-02)the Gansu beef cattle quality fattening project (GSAXMLZ-2021-01)+1 种基金the Application of Pingliang Red Bull Planting and Breeding Combined with High-efficiency Circular Production System Construction Technology Application (2020C-08)the local funding (GSSLCSX-2020-1)。
文摘Background Many countries have already banned the use of antibiotics in animal husbandry,making it extremely difficult to maintain animal health in livestock breeding.In the livestock industry,there is an urgent need to develop alternatives to antibiotics which will not lead to drug resistance on prolonged use.In this study,eighteen castrated bulls were randomly divided into two groups.The control group(CK)was fed the basal diet,while the antimicrobial peptide group(AP)was fed the basal diet supplemented with 8 g of antimicrobial peptides in the basal diet for the experimental period of 270 d.They were then slaughtered to measure production performance,and the ruminal contents were isolated for metagenomic and metabolome sequencing analysis.Result The results showed that antimicrobial peptides could improve the daily weight,carcass weight,and net meat weight of the experimental animals.Additionally,the rumen papillae diameter and the micropapillary density in the AP were significantly greater than those in the CK.Furthermore,the determination of digestive enzymes and fermentation parameters showed that the contents of protease,xylanase,andβ-glucoside in the AP were greater than those in the CK.However,lipase content in the CK was greater than that in the AP.Moreover,the content of acetate,propionate,butyrate,and valerate was found to be greater in AP than those in CK.The metagenomic analysis annotated 1993 differential microorganisms at the species level.The KEGG enrichment of these microorganisms revealed that the enrichment of drug resistance-related pathways was dramatically decreased in the AP,whereas the enrichment of immune-related pathways was significantly increased.There was also a significant reduction in the types of viruses in the AP.187 probiotics with significant differences were found,135 of which were higher in AP than in CK.It was also found that the antimicrobial mechanism of the antimicrobial peptides was quite specific.Seven low-abundance microorganisms(Acinetobactersp.Ac1271,Aequorivita soesokkakensis,Bacillus lacisalsi,Haloferax larsenii,Lysinibacillussp.3DF0063,Parabacteroidessp.217,Streptomycessp.So13.3)were found to regulate growth performance of the bull negatively.Metabolome analysis identified 45 differentially differential metabolites that significantly different between the CK and the AP groups.Seven upregulated metabolites(4-pyridoxic acid,Ala-Phe,3-ureidopropionate,hippuric acid,terephthalic acid,L-alanine,uridine 5-monophosphate)improve the growth performance of the experimental animals.To detect the interactions between the rumen microbiome and metabolism,we associated the rumen microbiome with the metabolome and found that negative regulation between the above 7 microorganisms and 7 metabolites.Conclusions This study shows that antimicrobial peptides can improve the growth performance of animals while resisting viruses and harmful bacteria and are expected to become healthy alternatives to antibiotics.We demonstrated a new antimicrobial peptides pharmacological model.We demonstrated low-abundance microorganisms may play a role by regulating the content of metabolites.
文摘The objectives of the study were to examine the effects of draught work on carcass characteristics, meat yield potential, and distribution along primal cuts in Hararghe highland bulls fed on net energy requirement basis. Twelve bulls were sorted into three groups of four animals each and assigned to three treatment hours: control (0), 4, or 6, using a complete randomized design (CRD). The results of the study show that there are no significant differences (p > 0.05) in slaughter weight, hot carcass weight (HCW), chilling loss, total edible and non-edible offal, fat thickness (FT), deboned meat yield (kg), meat to bone ratio, muscle distribution, and proportion of kidney, pelvic, and heart fat (KPH%) among the treatment groups. However, the hot carcass-based dressing percentage was significantly lowered (p < 0.05) in bulls that work 4 or 6 hours a day. Meat yield was strongly correlated with HCW (r = 0.74) and FT (r = 0.73). Therefore, the number of hours the bull spent on draught work did not negatively impact meat yield or quality. Consequently, the research suggests that utilizing Hararghe highland bulls for draught service, four or six hours a day is viable for dual advantages: crop cultivation and marketable carcass.
基金supported by a grant from the Natural Science Funding Projects of Shanxi Province,China(201801D121241)the Animal Husbandry Dominant Key Discipline Construction Project in“1331 Project”of Shanxi Province,China。
文摘This study evaluated the effects of palm fat powder(PFP) and coated folic acid(CFA) on growth performance, ruminal fermentation, nutrient digestibility, microbial enzyme activity, microflora, hepatic lipid content and gene expression in dairy bulls. Forty-eight Chinese Holstein bulls((362±12.4) days of age and(483±27.1) kg of body weight(BW)) were assigned to four groups in a completely randomized design with a 2×2 factorial arrangements. Supplemental PFP(0 or 30 g PFP kg-1 dietary dry matter(DM)) and CFA(0 or 120 mg FA d-1 as CFA) were mixed into the top one-third of a total mixed ration. The study included a 20-day adaptation period and followed by a 90-day collection period. The lower(P<0.01) feed conversion ratio with PFP or CFA addition resulted from the constant DM intake and the higher(P<0.05) average daily gain. The higher(P<0.05) ruminal p H, ether extract digestibility, microbial α-amylase activity, Butyrivibrio fibrisolvens copy, and expression of peroxisome-proliferator-activated receptor α(PPARα) and carnitine palmitoyl transferase-1(CPT1), and lower ruminal total volatile fatty acids(VFA) concentration, acetate to propionate ratio, neutral detergent fibre(NDF) digestibility, copies of total protozoa and Ruminococcus flavefaciens, and expression of sterol regulatory element binding protein-1(SREBP1) and acetyl-coenzyme A carboxylase α(ACACA) were observed for PFP addition. Supplementation with CFA increased(P<0.05) ruminal total VFA concentration, acetate to propionate ratio, digestibility of DM, organic matter, crude protein and NDF, activity of cellobiase, pectinase and α-amylase, copies of selected microbial except for total protozoa, as well as expression of PPARα, but decreased(P<0.05) ruminal p H, and expression of SREBP1 and ACACA. The PFP×CFA interaction(P<0.05) was observed for ammonia N, hepatic TG content, and m RNA expression of CPT1 and FAS. There had no significant difference in hepatic TG content when CFA was supplemented in the diet without PFP addition, the lower(P=0.001) hepatic TG content was observed when CFA was supplemented in the diet with PFP addition. The higher(P<0.05) m RNA expression of CPT1, and the lower(P<0.05) m RNA expression of FAS and ammonia N concentration were observed when CFA was supplemented in diet either without or with PFP addition. The results indicated that supplementation of CFA in PFP diet was more effective on increasing hepatic CPT1 expression, and decreasing ammonia N, hepatic TG content and FAS expression than in diet without PFP. Supplementation with PFP or CFA improved growth performance of dairy bulls by promoting nutrient utilization, microbial enzyme activity, microflora, and hepatic gene expression.
基金supported by the Major Project of National Transgene in China(2018ZX08007001-002)the National Natural Science Foundation of China(31671286,31672397,31771374)+2 种基金Shandong Provincial Natural Science Foundation for Distinguished Young Scholars of China(JQ201709)the Program of National Cow Industrial Technology System of China(CARS-37)the Shandong Provincial Key Research and Development Program of China(2017GNC10120).
文摘F-box and WD-40 domain protein 11(FBXW11)is an important component of the E3 ubiquitin-ligase enzyme that plays a key role in the ubiquitin-dependent regulation of spermatogenesis.In our previous research,the mRNA expression of FBXW11 in bull sperm with high motility is significantly higher than that with low motility.In the present study,the protein expression levels of FBXW11 in bull testicular tissues with low-performance sperm quality groups were significantly higher than those in normal performance groups.The immunohistochemistry result demonstrated that FBXW11 protein was located in the periphery of Leydig cells and seminiferous tubules.Three splice variants of the FBXW11 gene,namely,FBXW11-tv1,FBXW11-tv2,and FBXW11-tv3,were identified in testicular tissues.The splicing patterns of the three variants are exon skipping.The transcript FBXW11-tv2 expressions were the highest in each sample.The low-performance groups displayed higher FBXW11-tv1 and FBXW11-tv2 transcript expressions than the normal performance groups.Two CpG islands were located within the 5’UTR and exon 1-2 region of the FBXW11 gene.Bisulfite sequencing PCR results demonstrated that the methylation levels of 11 methylation sites in the CpG island 2 from−99 to−43 in the normal performance groups were significantly lower than those in the low-performance groups.Pearson correlation analysis suggested that the CpG island 2 methylation level was negatively correlated with sperm motility and the transcript FBXW11-tv2 expression level.Our data revealed that alternative splicing and DNA methylation jointly regulated FBXW11 gene expression and were correlated with sperm quality traits during spermatogenesis in Holsteins.
文摘Apple bull’s eye rot is caused by pathogenic Neofabraea species including N. malicorticis, N. perennans and N. alba. Fruits carrying this fungal quarantine disease are prohibited from entering China. The host plants for the pathogens include several Rosaceae fruits including apple and pear. Disease symptoms and pathogen morphology are often insufficient to determine the identity of the pathogen, particularly at the species level. In the current study, we analyzed the inter-species sequence variations in the β-tublin gene, and designed specific primers to allow PCR amplification of 554 bp fragments from pathogenic Neofabraea species. The PCR products were recovered and sequenced, and Blast search was conducted using the DNA sequences in the Genbank database. The results indicated precise PCR amplification of the target sequences from the host pathogen, which allowed unambiguous identification of the species.
文摘Osteopontin (OPN) is a protein found at higher concentrations in the seminal plasma of bulls with above average fertility. Polymorphisms have been reported within the OPN gene promoter that can affect production of this protein and thus, affect fertility. Therefore, Angus (n = 5) and Angus x Gelbvieh (Balancer, n = 14) and Angus x Brahman (n = 15) bulls were evaluated for presence of single nucleotide polymorphisms (SNP) in the Bos taurus OPN gene (GenBank: AY878328.1) promoter region, and their possible effects on bull semen quality as evaluated by computer-assisted semen analysis (CASA). Semen was collected by electroejaculation 6 to 9 times from each bull, and each semen collection was evaluated by CASA for motile, progressive and rapid sperm within 5 mins of ejaculation. The bulls were genotyped for reported single nucleotide polymorphisms (SNP) in the promoter region of the OPN gene through amplification of two 700 base pair (bp) DNA fragments and sequencing of the resulting PCR products. Seven SNP sites were identified, at bp 3379, 3490, 3492, 5075, 5205, 5209, and 5263 of the OPN gene. The SNP identified at bp 5205, 5209 and 5263 had not been previously reported. Individual SNP sites were evaluated as the main effect on CASA sperm motility variables in a SAS MIXED model for repeated measures. A thymine to guanine substitution at bp 3379 was associated with increased (P ≤ 0.02) percentage of motile, progressive and rapid sperm in Angus x Brahman bulls, and tended (P ≤ 0.10) to increase the same sperm motility parameters in Angus, and Angus x Gelbvieh bulls. The percentages of motile, progressive and rapid sperm were similar (P ≥ 0.05) among genotypes for the other 6 SNP identified. These results suggest that identification and genotyping of polymorphisms within the promoter region of the bovine OPN gene may be useful for selecting bulls with improved sperm motility parameters.
基金the Provincial Breeders Federation of Trento for their support
文摘Background: Semen evaluation is used to estimate the testicular function. In bulls, the spermatozoa present in the ejaculate are the result of a process that begun more than 2 mo earlier, bequeathing a delayed depiction of the actual function of the testis. Since testis vascularization might be critical for the gonad function, selected pulse wave Doppler ultrasound parameters were assessed in this study, for instance the peak systolic velocity, the end diastolic velocity and the resistive index of the testicular artery along the spermatic cord, the marginal portion of the testicular artery and the intratesticular branches of the testicular artery both in healthy adult and young bulls.Correlations between these parameters and characteristics of semen that was collected numerous times, before and after the Doppler ultrasound examination.Results: The peak systolic velocity and the end diastolic velocity measured in the testicular artery along the spermatic cord(supratesticular artery – SA) were variable among the bulls and within individual bulls, likely due to the convoluted course of the vessel. The resistive index was found highly repeatable in the same bull. A reduction in the resistive index was found between the supratesticular artery and the marginal portion of the testicular artery(P < 0.01), and between the marginal portion of the testicular artery and the intratesticular branches of the testicular artery(P < 0.05). No differences were recorded for the pulse wave Doppler ultrasound parameters in young bulls compared with adults. A significant correlation was found between the resistive index of the marginal portion of the testicular artery and total sperm in the ejaculate(r = 0.516, P < 0.05), the immature sperm(r = 0.462, P < 0.05), the teratoid sperm(r = 0.375, P < 0.05), and the "Dag defect" sperm(r = 0.389, P < 0.05). Similarly, the resistive index of the intratesticular branches of the testicular artery were found correlated with the total sperm number in the ejaculate(r = 0.568, P < 0.05), the immature sperm(r = 0.523, P < 0.05), the teratoid sperm(r = 0.418, P < 0.05), and the "Dag defect" sperm(r = 0.341, P < 0.05).Conclusions: The data presented in this study suggest that the resistive index, measured at the marginal portion of the testicular artery, could be an easy-to-perform parameter to evaluate the spermatogenesis quality in young bulls and normal adults.
基金Supported by the National Key Special Science and Technology Project(2016ZX05015-002)PetroChina Key Special Science and Technology Project(2016E-0104)
文摘Aimed at the disadvantages of secondary damage to oil layers caused by the conventional bull-heading water control technique, a thermo-sensitive temporary plugging agent for water control was synthesized by water solution polymerization and applied in the field with a new secondary temporary plugging technique. The optimization and performance evaluation of thermo-sensitive temporary plugging agent were carried out through laboratory experiments. The optimized formula is as follows:(6%-8%) acrylamide +(0.08%-0.12%) ammonium persulfate +(1.5%-2.0%) sepiolite +(0.5%-0.8%) polyethylene glycol diacrylate. The thermo-sensitive temporary plugging agent is suitable for formation temperatures of 70-90 ?C, it has high temporary plugging strength(5-40 k Pa), controllable degradation time(1-15 d), the apparent viscosity after degradation of less than 100 m Pa?S and the permeability recovery value of simulated cores of more than 95%. Based on the research results, secondary temporary plugging technique was used in a horizontal well in the Jidong Oilfield. After treatment, the well saw a drop of water cut to 27%, and now it has a water cut of 67%, its daily increased oil production was 4.8 t, and the cumulative oil increment was 750 t, demonstrating that the technique worked well in controlling water production and increasing oil production.
基金the support from the Ministry of ScienceInnovation and Universities,Spain (AGL2017–88329-R and FPU18/00666)+1 种基金Regional Government of Catalonia,Spain (2017-SGR-1229)University of Girona (Postdoc-Ud G2020)。
文摘Background:Genetic selection in cattle has been directed to increase milk production.This,coupled to the fact that the vast majority of bovine artificial inseminations(AI)are performed using cryopreserved sperm,have led to a reduction of fertility rates over the years.Thus,seeking sensitive and specific sperm biomarkers able to predict fertility rates is of vital importance to improve cattle reproductive efficiency.In humans,sperm chromatin condensation evaluated through chromomycin A3(CMA3)has recently been purported to be a powerful biomarker for sperm functional status and male infertility.The objectives of the present study were:a)to set up a flow cytometry method for simultaneously evaluating chromatin condensation and sperm viability,and b)to test whether this parameter could be used as a predictor of in vivo fertility in bulls.The study included pools of three independent cryopreserved ejaculates per bull from 25 Holstein males.Reproductive outcomes of each sire were determined by non-return rates,which were used to classify bulls into two groups(highly fertile and subfertile).Results:Chromatin condensation status of bovine sperm was evaluated through the combination of CMA3 and Yo-Pro-1 staining and flow cytometry.Sperm quality parameters(morphology,viability,total and progressive motility)were also assessed.Pearson correlation coefficients and ROC curves were calculated to assess their capacity to predict in vivo fertility.Sperm morphology,viability and total motility presented an area under the ROC curve(AUC)of 0.54,0.64 and 0.68,respectively(P>0.05),and thus were not able to discriminate between fertile and subfertile individuals.Alternatively,while the percentage of progressively motile sperm showed a significant predictive value,with an AUC of 0.73(P=0.05),CMA3/Yo-Pro-1 staining even depicted superior results for the prediction of in vivo fertility in bulls.Specifically,the percentage of viable sperm with poor chromatin condensation showed better accuracy and precision to predict in vivo fertility,with an AUC of 0.78(P=0.02).Conclusions:Chromatin condensation evaluated through CMA3/Yo-Pro-1 and flow cytometry is defined here as a more powerful tool than conventional sperm parameters to predict bull in vivo fertility,with a potential ability to maximising the efficiency of dairy breeding industry.
基金Supported by NMKJ Project of Production and Industrial Application of Sexed Semen in Domestic Animals(No.20111701)
文摘[Object]This study aimed to examine the effects of 3,7-dimethyl-1-(5-oxo-hexyl)-xanthine(pentoxifylline,PF)on the motility,mitochondrial activity,acrosome integrity and fertilization rate of spermatozoa in both fresh and frozen-thawed bull semen.[Method]Fresh and frozen-thawed bull spermatozoa were exposed to 5 mmol/L PF with untreated samples as controls.[Result]Fresh spermatozoa showed reduced(P<0.05)motility after 2 h incubation with PF whereas,surprisingly,frozen-thawed samples exhibited increased sperm motility(P<0.05)after 2 h incubation with PF and they also showed enhanced longevity compared to controls.Mitochondrial activity in both fresh and frozen-thawed bull spermatozoa increased(P<0.05)during 4 h incubation with PF whereas acrosome integrity remained unchanged in both types of semen.However,treatment with 5 mmol/L PF did not influence the in vitro fertilization efficiency of fresh spermatozoa but improved significantly(P<0.05)that of frozen-thawed spermatozoa.[Conclusion]These results indicate that PF can improve sperm quality of frozenthawed bull semen,and may improve pregnancy rates in bovine artificial insemination programmes employing frozen semen.
文摘Objective:To evaluate the effect of adding different values of polymyxin B (PMB) to bull semen on various motility parameters of post-thawed semen such as total motility, progressive motility and velocity parameters using kinetic parameters of sperm by Computer Assisted Sperm Analysis.Methods: Gram negative bacteria release lipopolysaccharide, which induces the apoptotic pathway. Antibiotics are added to semen in order to prevent bacterial contaminations in bovine semen. These antibiotics kill the bacteria especially gram negative bacteria. Therefore, their endotoxins are released during bacteriolysis and bind to the head region and midpiece of sperm. PMB is a bactericidal antibiotic against multidrug resistant gram-negative bacteria and is able to neutralize the toxic effects of the released endotoxin. This study was performed on 3-year old Taleshi bulls. Results:The results showed both positive and negative significant effects of PMB on semen quality. Total motility and progressive motility were significantly increased (P<0.0001) by 100 μg per mL of PMB (55.2% and 48.8% respectively) against the control groups (43.5% and 37.7%, respectively). Moreover, they were significantly decreased (P<0.0001) by 1000 μg per mL of PMB (35.2% and 28.8% respectively) against the control groups (43.5% and 37.7% respectively) in above-mentioned parameters. In Computer Assisted Semen Analyzer, parameter VAP was significantly decreased (P<0.04) in 1000 μg (69.6 μm/s) against the control group (78.7 μm/s). Finally, using PMB in processing cryopreserved bull semen is advised, but before using it, the rate of endotoxins must be measured.Conclusions: We advise using PMB after measuring endotoxin concentration;In vitro,in vivoand in field fertilization, adding other sperm evaluation factors such as acrosomal integrity, DNA integrity, mitochondrial function to PMB treated semen.
文摘The objective of the study was to determine the level of Bruising and Dark Firm Dry on Arsi, Boran and Harar cattle breeds in Ethiopia. A total of 39 beef bulls from three different breeds (14 Boran, 14 Harar, and 11 Arsi cattle breeds) were used in this study. Based on the present finding, from 39 slaughtered beef bulls 66.7% was bruised and 30.77% produced DFD meat. Harar breed had the highest (39.1%) overall level of bruising while Boran and Arsi had the lowest level. However, Harar breed had the lowest level of DFD (14.29%). Age groups 3 - 6 and 6 - 9 years had the highest level of a bruise (29.70% and 28.10%, respectively) compared to age groups 2 - 3 years. The mean number of bruises per carcass was 1.96 ± 0.25 while the bruise score per carcass was 3.58 ± 0.68. A linear bruise was the most observed (33.33%) shape of a bruise. It was concluded that the level of bruising and DFD was higher compared to some other abattoir studies in other countries which indicated the possible presence of poor handling practice of cattle from farm to abattoir gates. To minimize the incidence of DFD and bruising, it was recommended that proper handling of cattle during loading, transport, unloading, at lairriage and during slaughter need to be practiced.
文摘Data consisting of test performance records of 725 bulls was analysed to evaluate trends for growth and feed efficiency, and to determine environmental factors that influence growth of range performance tested bulls in the arid sweet bushveld of South Africa. Performance data were subjected to regression procedures to estimate parameters of the slope (β) and intercept (α) for average daily gain (ADG) and Kleiber ratio (KR). Mixed procedures were applied to test for sources of variation in ADG and KR. There was an increase of 3.481 g/day/year and 0.528 g/day/ year in ADG for Nguni and Bonsmara bulls, respectively. The ADG trends’ respective feed efficiency as reflected by KR was 0.093 and 0.059. Herd of origin and test-year were the sources of variation for ADG while the variation in KR was due to the herd of origin, test-year, start-age and start-weight. Results indicated a steady increase in ADG and KR for range performance tested bulls in the arid sweet bushveld.
文摘The aim of this study was to assess sperm quality (motility, viability and acrosomal integrity) sperm from commercially frozen semen straws two breeds of bulls Bos taurus (holstein Frisian) and Bos indicus (Brahman). 9 commercial straws 0.5 ml of Holstein bull semen and 9 Brahman bull were thawed, they were kept for two hours at room temperature and motility, viability and acrosomal integrity (NAR) was assessed. The results were 30% motility, viability 40% and 30% of NAR in the Holstein breed. Brahma race for motility 40%, 50% and 40% viability was obtained NAR. In conclusion, according to the results of the variables analyzed, the Brahman breed in sperm quality was better than the Holstein breed;however, the results of both races meet minimum standards of quality sperm for use in artificial insemination (AI) field level.
文摘The study was planned with an objective to assess the level of antisperm antibodies (ASA) in the blood serum and seminal plasma of breeding cow bulls and their relationship with sperm function and fertility tests. ASA was analyzed in blood serum and seminal plasma by SpermMar test, Immuno peroxidase assay (IPA) and Enzyme linked immunoabsorbant assay (ELISA). In SpermMar test, about 54% bulls were with >40% IgG in blood serum against sperm surface antigens, whereas none of the bulls were with >10% IgG in seminal plasma. More than 20% and >10% IgA against sperm surface antigens were detected in the blood serum and seminal plasma of 65.8% and 37% bulls, respectively. Out of 26 bulls, seminal plasma of 21 bulls reacted with spermatozoa both in IPA and IgA latex particles and that of only 12 bulls reacted with IgG. In IPA, about 50% of the bulls had >40% ASA against head surface antigens, whereas, there were 23% bulls with >10% ASA in seminal plasma. Also ELISA indicated a higher antibody titre in blood serum (3200 - 6400) and seminal plasma (40 - 80) of 50% and 42% bulls, respectively. There were 11 bulls with low values of HOST/in vitro acrosome reaction/cervical mucus penetration assay and higher level of either serum or seminal plasma ASA. Our study revealed that a significant level of ASA in serum or seminal plasma may have effect on the fertility of bulls by affecting the sperm function.
文摘Seminal plasma is composed of secretions from accessory sex glands, which are mixed with sperm at ejaculation and contribute the majority of semen volume. Seminal plasma is considered a transport and support medium for sperm in the female reproductive tract. Because seminal plasma is not required for fertilization, the importance of its constituents to the establishment of normal pregnancy has been overlooked. Four seminal plasma proteins, Osteopontin, Sper-madhesin Z13, BSP 30 kDa and Phospholipase A2, have been identified as markers of fertility in dairy bulls (Cancel et al., 1997;Moura et al., 2006, 2007). The objective of the present study was to characterize the expression patterns of these proteins and other proteins found to be of interest in seminal plasma of cryopreserved and non-cryopreserved bull semen. Seminal plasma samples were obtained from 16 mature Hol-stein-Friesian bulls at Select Sires Inc. Samples were divided into two groups based on assigned fertility score expressed as the percentage point deviation (PD) of the bull’s non-return rate (NRR) from the average NRR of all bulls in the Select Sires Inc. reproductive management program. Group 1 (high fertility bulls, n = 8) 1.9 ≤ PD ≤ 2.7%, and group 2 (low fertility bulls, n = 8) -6.5 ≤ PD ≤ 1.8 %. Additionally, the samples were categorized as processed (cryopreserved) or unprocessed (non-cryopreserved) for protein analysis. Protein expression was analyzed by 2-D fluorescence difference gel electrophoresis (2D-DIGETM). Protein spots were picked from a reference gel, analyzed by mass spectrometry and, subsequently identified by MS/MS ion searches performed on the SwissProt database. Protein expression did not differ (P > 0.05) with fertility grouping but displayed two distinct patterns among the processing groups: majority of the functional proteins were highly expressed in seminal plasma of non-cryopreserved semen while the cryopreserved semen contained mainly structural/extender derived proteins. Functional proteins identified included Spermadhesin Z13, BSP A1/2, BSP 30 kDa, Nucleobindin-1 and metalloproteinase inhibitor 2. Some of these proteins have been identified as anti-fertility or fertility enhancing agents in males. Whether this alteration in protein expression after processing might affect semen fertility is worthy of further evaluation.
文摘Objective:To optimize the cryopreservation of buffalo bull semen by altering freezing rates within critical temperature range (4℃ to -60℃). Methods: A total of 20 ejaculates each from 5 Murrah buffalo bulls were cryopreserved using programmable biofreezer in 2 phases. In the 1st phase, 9 freezing rates were applied at -2, -5, -10, -20, -30, -40, -50, -60 or -4℃/min (control) from 4℃ to -15℃;at -40℃/min from -15℃ to -60℃. In the 2nd phase, a fixed freezing rate was applied at -30℃/min from 4℃ to -15℃. Six freezing rates were applied at -10, -20, -30, -40 (control), -50 or -60℃/min from -15℃ to -60℃. The freezing from -60℃ to -140℃ were fixed at -50℃/min in both the phases. Post thaw semen quality was assessed in terms of motility, viability, membrane integrity (hypo-osmotic swelling test), sperm abnormalities, and active mitochondria. Data were arc sine transformed and analyzed through one-way analysis of variance using SPSS software. Results: In the 1st phase, percent individual motility, progressive motility and viability were similar among various protocols. Percent hypo-osmotic swelling reactive sperm was higher with freezing at -30℃/min. In the 2nd phase, percent individual motility, viability and hypo-osmotic swelling reactive sperm was higher with freezing at -50℃/min. Sperm head abnormalities were lower at -30℃/min in the 1st phase, but were similar among the protocols of the 2nd phase. Percent active mitochondria were higher at -30℃/min in the 1st phase and at -50℃/min in the 2nd phase.Conclusions:The optimum post thaw semen quality of buffalo bull could be obtained by applying freezing rate at -30℃/min (4℃ to -15℃) and at -50℃/min (-15℃ to -140℃), followed by plunging of straws in into liquid nitrogen for storage.
文摘Objective:To evaluate the supplementation effects of vitamin E,vitamin C,superoxide dismutase(SOD),and catalase to diluents on bull cryopreserved epididymal sperm.Methods:Sperm were retrieved from 20 bull testes and were then supplemented with 0.1 mM vitamin E,5.0 mM vitamin C,100.0 IU/mL SOD,and 100.0μg/mL catalase alone,or in a combination.The control treatment contained no addition.After supplementation,samples were frozen and stored in liquid nitrogen.The sperm parameters including motility,progressive motility,viability,acrosome integrity,plasma membrane integrity,kinematics and DNA damage were evaluated following the thawing process.Results:Vitamin E alone significantly increased the parameters of acrosome and membrane integrity compared to the control treatment(P<0.05).While compared to the control treatment,vitamin C had no improvement effect on sperm characteristics except for membrane integrity.Treatment of vitamin E+vitamin C had a significant improvement in total motility,progressive motility,viability,membrane and acrosome integrity compared to the control and other treatments(P<0.05).Compared to the control treatment,addition of SOD or catalase alone significantly improved the percentages of total motility,progressive motility,viability,membrane and acrosome integrity(P<0.05).Furthermore,SOD+catalase significantly increased total motility,progressive motility,viability,acrosome and membrane integrity characteristics compared to the catalase treatment(P<0.05).Vitamin E alone,vitamin E+vitamin C,and SOD in diluents decreased DNA damages and thereby improved the rate of intact sperm heads.Conclusions:Addition of 100.0 IU/mL SOD alone and 0.1 mM vitamin E+5.0 mM vitamin C,and also 5.0 mM vitamin C+100μg/mL catalase in a combination improves the quality of cryopreserved bull epididymal sperm and could be used for cryopreservation.
文摘Objective:To assess the hemodynamic changes that occurred in the main testicular artery at two different locations such as supra and marginal ones in bulls at three different ages.Methods:Eighteen Baladi bulls were divided according to their age into three different age groups:1 year[n=6;groupⅠwith bodyweight(380±10)kg],3 years[n=6;groupⅡwith bodyweight(570±10)kg],and 6 years old[n=6;groupⅢwith bodyweight(650±10)kg]Baladi bulls.Circumference of the scrotum,thickness of the mediastinum,testicular dimensions,vascularity,and hormonal levels were measured.Testicular hemodynamics were assessed by Doppler ultrasound scanning.Results:Testicular width,length,and volume were significantly different among the three age groups,with the highest in bulls of groupⅢ(P<0.05).The bulls in groupⅢhad significantly higher mediastinal line thickness and scrotal circumference than groupⅠ(P<0.05),but there was no significant difference between groupⅡand groupⅢ.Bulls in groupⅡshowed a decline in Doppler indices(resistance and pulsatility indexes)at both testicular branches compared to the other two groups(P<0.05).Supra and marginal end diastolic velocities in the three age groups were significantly different,with the highest in groupⅡ(P<0.05).Time average velocity(cm/s)was significantly elevated in groupⅡcompared to the other two groups(P<0.05).Both testicular echotextures were significantly decreased in groupⅡ(P<0.05).Estradiol and nitric oxide metabolites were significantly high in groupⅡcompared to the other two groups(P<0.05).Meanwhile,the level of plasma testosterone was maximum in groupⅢ(P<0.05).Positive correlations were found between supratesticular artery resistance and pulsatility indexes(r=0.81;P<0.001),while both resistance and pulsatility indexes had correlated negative with estradiol(r=-0.71 and r=-0.91;P<0.001),and nitric oxide metabolites(r=-0.92 and r=-0.72;P<0.001).Conclusions:Three-year old Baladi bull has the lesser Doppler indices with lesser echotexture and greater nitric oxide with estradiol concentrations,which directs a significant ability for the thermoregulation process.These parameters will help in breeding selection.
文摘BACKGROUND Hepatocellular adenomas are rare tumors that can occur in patients with glycogen storage disease type I.CASE SUMMARY We herein report two cases of histologically proven hepatocellular adenomas in patients with glycogen storage disease type I.Magnetic resonance imaging(MRI)was performed after bolus injection of gadoxetate disodium,a liver-specific gadolinium-based MRI contrast agent.In the present cases,some of the hepatocellular adenomas showed unexpectedly a“bull’s eye”appearance on T2-weighted and post-contrast images,which was not previously described as imaging findings of hepatocellular adenomas in glycogen storage disease.A bull’s eye appearance on T2-weighted images can be encountered in both benign(i.e.,abscess)or malignant(i.e.,epithelioid hemangioendothelioma,cholangiocarcinoma,and metastases)hepatic lesions.CONCLUSION We present two cases of hepatocellular adenomas in patients with glycogen storage disease type 1,in which gadoxetate disodium-MRI showed atypical imaging findings for hepatocellular adenomas.At present there is no systematic study evaluating MRI findings of hepatocellular adenomas in patients with glycogen storage disease,further studies are needed to specifically investigate this issue.
