An illness known as pneumonia causes inflammation in the lungs.Since there is so much information available fromvarious X-ray images,diagnosing pneumonia has typically proven challenging.To improve image quality and s...An illness known as pneumonia causes inflammation in the lungs.Since there is so much information available fromvarious X-ray images,diagnosing pneumonia has typically proven challenging.To improve image quality and speed up the diagnosis of pneumonia,numerous approaches have been devised.To date,several methods have been employed to identify pneumonia.The Convolutional Neural Network(CNN)has achieved outstanding success in identifying and diagnosing diseases in the fields of medicine and radiology.However,these methods are complex,inefficient,and imprecise to analyze a big number of datasets.In this paper,a new hybrid method for the automatic classification and identification of Pneumonia from chest X-ray images is proposed.The proposed method(ABOCNN)utilized theAfrican BuffaloOptimization(ABO)algorithmto enhanceCNNperformance and accuracy.The Weinmed filter is employed for pre-processing to eliminate unwanted noises from chest X-ray images,followed by feature extraction using the Grey Level Co-Occurrence Matrix(GLCM)approach.Relevant features are then selected from the dataset using the ABO algorithm,and ultimately,high-performance deep learning using the CNN approach is introduced for the classification and identification of Pneumonia.Experimental results on various datasets showed that,when contrasted to other approaches,the ABO-CNN outperforms them all for the classification tasks.The proposed method exhibits superior values like 96.95%,88%,86%,and 86%for accuracy,precision,recall,and F1-score,respectively.展开更多
A cross-sectional study was conducted to determine the Seropositivity of Toxoplasma gondii in water buffalo in three Iraqi governorates (Baghdad, Dhi Qar, and Maysan) and to estimate the risk aspects related to infest...A cross-sectional study was conducted to determine the Seropositivity of Toxoplasma gondii in water buffalo in three Iraqi governorates (Baghdad, Dhi Qar, and Maysan) and to estimate the risk aspects related to infestation throughout the period from January to December 2019. A total of 430 serum samples were inspected with a commercial ELISA (Enzyme linked immunosorbent assay) kit. Indirect multi-species kit. The overall Seropositivity of T. gondii in the examined local buffalo was 7.4%, and the highest rate (9.3%) was in Baghdad Governorate. A multivariate regression analysis revealed that adult buffalo (OR = 7.10;95% CI: 0.87-57.68;P = 0.067) and young herds (OR = 8.42;95% CI: 1.07-66.02;P = 0.043) were more subject to infestation from young buffalo and large herds. Furthermore, the hazard of toxoplasmosis was increased in winter especially among animals in contact with cats. It is therefore requisite to determine risk aspects to evaluate which mitigation, control, prevention and procedures should be carried out to diminish, control and prevent infestation with T. gondii and its propagation.展开更多
Background During mammalian pre-implantation embryonic development(PED),the process of maternal-to-zygote transition(MZT)is well orchestrated by epigenetic modification and gene sequential expression,and it is related...Background During mammalian pre-implantation embryonic development(PED),the process of maternal-to-zygote transition(MZT)is well orchestrated by epigenetic modification and gene sequential expression,and it is related to the embryonic genome activation(EGA).During MZT,the embryos are sensitive to the environment and easy to arrest at this stage in vitro.However,the timing and regulation mechanism of EGA in buffaloes remain obscure.Results Buffalo pre-implantation embryos were subjected to trace cell based RNA-seq and whole-genome bisulfite sequencing(WGBS)to draw landscapes of transcription and DNA-methylation.Four typical developmental steps were classified during buffalo PED.Buffalo major EGA was identified at the 16-cell stage by the comprehensive analy-sis of gene expression and DNA methylation dynamics.By weighted gene co-expression network analysis,stage-spe-cific modules were identified during buffalo maternal-to-zygotic transition,and key signaling pathways and biological process events were further revealed.Programmed and continuous activation of these pathways was necessary for success of buffalo EGA.In addition,the hub gene,CDK1,was identified to play a critical role in buffalo EGA.Conclusions Our study provides a landscape of transcription and DNA methylation in buffalo PED and reveals deeply the molecular mechanism of the buffalo EGA and genetic programming during buffalo MZT.It will lay a foundation for improving the in vitro development of buffalo embryos.展开更多
Objective:To optimize the concentration of PDC-109 protein in semen for higher cryopreservability and fertility by sequestration or neutralization of PDC-109 by its antibodies(anti-PDC-109 antibodies)in bubaline speci...Objective:To optimize the concentration of PDC-109 protein in semen for higher cryopreservability and fertility by sequestration or neutralization of PDC-109 by its antibodies(anti-PDC-109 antibodies)in bubaline species.Methods:PDC-109 protein was purified by applying two-step chromatography procedures.Purified protein was injected in rabbits to raise antibodies.These raised anti-PDC-109 antibodies were used in neutralization or sequestration of PDC-109 in in-vitro model.Ejaculates were collected from buffaloes and splited for four groups.Group 1 received egg yolk Tris glycerol extender,without anti-PDC-109 antibodies,while group 2 to 4 received anti-PDC-109 antibodies 266μg/mL,80μg/mL,and 26μg/mL in Tris-fructose-citrate buffer,respectively.Semen quality parameters viz.,forward progressive motility,viability,total morphological abnormality,acrosomal integrity,plasma membrane integrity,cryoinjury and in-vitro zona binding index were evaluated.Results:Semen quality parameters of neat semen were within the normal range of bubaline species.Sperm motility,livability,acrosomal integrity,plasma membrane integrity,and cholesterol content of sperm were decreased and total sperm abnormality was increased significantly in post-thaw semen compared to those in pre-freeze and fresh semen(P<0.05).Semen in group 2 had higher sperm motility,livability,acrosomal integrity,plasma membrane integrity,and cholesterol content of sperm and lower total sperm abnormality significantly compared to those in group 1,3 and 4 at pre-freeze and post-thaw stages(P<0.05).Conclusions:Sequestration or neutralization of PDC-109 by its antibodies significantly improves pre-freeze,and post-thaw semen quality parameters and in-vitro zona binding index with simultaneously reducing cryoinjury or cryodamage in the sperm of bubaline species.展开更多
Software needs modifications and requires revisions regularly.Owing to these revisions,retesting software becomes essential to ensure that the enhancements made,have not affected its bug-free functioning.The time and ...Software needs modifications and requires revisions regularly.Owing to these revisions,retesting software becomes essential to ensure that the enhancements made,have not affected its bug-free functioning.The time and cost incurred in this process,need to be reduced by the method of test case selection and prioritization.It is observed that many nature-inspired techniques are applied in this area.African Buffalo Optimization is one such approach,applied to regression test selection and prioritization.In this paper,the proposed work explains and proves the applicability of the African Buffalo Optimization approach to test case selection and prioritization.The proposed algorithm converges in polynomial time(O(n^(2))).In this paper,the empirical evaluation of applying African Buffalo Optimization for test case prioritization is done on sample data set with multiple iterations.An astounding 62.5%drop in size and a 48.57%drop in the runtime of the original test suite were recorded.The obtained results are compared with Ant Colony Optimization.The comparative analysis indicates that African Buffalo Optimization and Ant Colony Optimization exhibit similar fault detection capabilities(80%),and a reduction in the overall execution time and size of the resultant test suite.The results and analysis,hence,advocate and encourages the use of African Buffalo Optimization in the area of test case selection and prioritization.展开更多
In the discipline of Music Information Retrieval(MIR),categorizing musicfiles according to their genre is a difficult process.Music genre classifica-tion is an important multimedia research domain for classification of mu...In the discipline of Music Information Retrieval(MIR),categorizing musicfiles according to their genre is a difficult process.Music genre classifica-tion is an important multimedia research domain for classification of music data-bases.In the proposed method music genre classification using features obtained from audio data is proposed.The classification is done using features extracted from the audio data of popular online repository namely GTZAN,ISMIR 2004 and Latin Music Dataset(LMD).The features highlight the differences between different musical styles.In the proposed method,feature selection is per-formed using an African Buffalo Optimization(ABO),and the resulting features are employed to classify the audio using Back Propagation Neural Networks(BPNN),Support Vector Machine(SVM),Naïve Bayes,decision tree and kNN classifiers.Performance evaluation reveals that,ABO based feature selection strategy achieves an average accuracy of 82%with mean square error(MSE)of 0.003 when used with neural network classifier.展开更多
The Dehong buffalo( D),F1 hybrids of Dehong buffalo × Murrah buffalo( MD) and F1 hybrids of Dehong × Nili buffalo( ND) as the experimental animals to investigate their fatty acid composition and amino acid c...The Dehong buffalo( D),F1 hybrids of Dehong buffalo × Murrah buffalo( MD) and F1 hybrids of Dehong × Nili buffalo( ND) as the experimental animals to investigate their fatty acid composition and amino acid content. Five of each buffalos were bred by freely grazing and slaughtered at 35. 8-month old age to investigate the fatty acid composition and amino acid content in beef of Dehong buffalo,MD buffalo and ND buffalo. The results showed that,saturated fatty acid content of Dehong buffalo( 44. 33%) was significantly higher than that of ND buffalo( 41. 39%) and MD buffalo( 41. 48%)( P 【 0. 05),and the latter two were not significantly different( P 】 0. 05). The content of PUFA in Dehong buffalo( 53. 90%) was significantly lower than ND( 56. 83%) and MD( 56. 83%)( P 【0. 05),however,the n- 6∶ n- 3( 4. 25) of Dehong buffalo was significantly higher than ND( 3. 14) and MD( 3. 52)( P 【 0. 05). The content of amino acids in muscle was not significantly different between Dehong buffalo,MD buffalo and ND buffalo( P 】 0. 05),while the acidic amino acid content of Dehong buffalo was higher than that of MD buffalo and ND buffalo.展开更多
[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying bio...[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.展开更多
To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochem...To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochemistry SuperPicTureTM two step method was used to detect the distribution of OT in hypothalamatic-pituitary-ovary axis of five femal Guangxi local buffalo. The test results could provide morphology according to study the OT's synthesis and mechanism of action,and could play reference and directions part in breeding Guangxi local buffalo. The test results display:oxytocin immuno reactive (OT-IR) neuronsw eremainly distributed arcuate nucleus,supraoptic nucleus and paraventricular nucleus,and OT-IR neurons was also found in ventromedial nucleus,ventrolateralis nucleus,suprachiasmaticus nucleus,dorsomedial nucleus,mamillary body,anterior hypothalamic nucleus and so on. The OT immunoactive production was found in pituitary and few OT-IR nerve fibers extended to post pituitary from hypophyseal stalk and medium eminence. In ovaries,OT immunoactive productions were only distributed in germinal epithelium cells,granulosa cells and lutein cells. The OT was first discovered in singulorum link of hypothalamatic-pituitary-ovary axis of Guangxi local buffalo. The OT immunoactive neurons were first discovered in every main nucleus of Guangxi local buffalo hypothalamus,especially distributed in arcuate nucleus,supraoptic nucleus and paraventricular nucleus.展开更多
[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of Buffalo. [Method] Buffalo oocytes were randomly divided into two groups 20-22 h after in ...[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of Buffalo. [Method] Buffalo oocytes were randomly divided into two groups 20-22 h after in vitro maturation. One group of oocytes was introduced with about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection 7-10 h or 18-20 h after in vitro fertilization (IVF); the other group of oocytes was introduced with mixture of a single buffalo sperm and about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection (generally called ICSI-Mediated Gene Transfer, ICSI-Tr). Expression of exogenous DNA was observed and recorded during the process of embryonic development. [Result] Early embryonic gene expression efficiency and blastocyst gene expression efficiency in IVF injection group showed no significant difference compared with that in ICSI-Tr group (P0.05). In addition, the cleavage rate and early embryonic gene expression efficiency in IVF injection group were significantly higher with injection at 7-10 h post IVF than that at 18-20 h post IVF (P0.05). [Conclusion] These results indicate that transgenic buffalo embryos can be generated by injecting exogenous DNA into cytoplasm of IVF oocytes, and the optimal injection time is 7-10 h post IVF.展开更多
The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out ...The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer (May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized (n=11) and control (n=13) groups. Each immunized cow was administered with a 2-mL immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibinα-subunit fusion protein. The controls were treated with the adjuvant only. Al animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artiifcial y inseminated upon behavioral estrus. As a result, al of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of fol icle-stimulating hormone (FSH), activin, and estradiol (E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls (72%vs. 30%, P<0.05). On aver-age, inhibin-immunized buffaloes had signiifcantly more large fol icles (≥9 mm in diameter) than the controls (mean±SEM;1.2±0.1 vs. 0.84±0.1, respectively;P<0.05) and a slightly higher mean total number of fol icles (≥2 mm;11.4±0.7 vs. 9.0±1.1, respectively;P=0.09) and smal (2–4 mm) fol icles (8.81±0.6 vs. 6.84±1.0, respectively;P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group (91%(10/11) vs. 54%(7/13), respectively;P<0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum (CL) than the controls 9 days after ovulation and signiifcantly higher (P<0.01) post-ovulation peak plasma progesterone (P4) concentrations. Immunization against inhibin also mar-ginal y increased the conception rate 42 days after insemination (45.8%vs. 15.4%;P>0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.展开更多
Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 -...Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 - 3 years old and weighing 300 - 500 kg as well as six goats were confirmed free of fasciolosis by fecal examination and Dot-ELISA. Two studies were conducted using the water buffaloes. In the first experiment, 8 water buffaloes were randomly divided into control group (n = 3) and infection group (n = 5). Each buffalo in the infected group received orally 60 metacercariae of F. hepatica per day for 20 days (total 1 200 metacercariae) to produce a chronic infection. In the second experiment, 12 water buffaloes were randomly divided into infected (n=9) and control group (n = 3). Each buffalo in the infected group was given a single oral dose of 1 600 metacercariae to produce an acute infection. The 6 goats were randomly divided into two infected groups and a control group. The sheep in two infections received a single oral dose of 200 and 500 metacercariae respectively, the control group remained uninfected. Blood NO and TNF-a concentrations of the test animals were measured by a reductive enzyme assay and RIA, respectively. Blood NO concentration in both acutely and chronically infected water buffaloes progressively increased from week 3 post-infection and was significantly greater than that of the control group (P<0. 05) at the 5th week (acute infection) and 7th week (chronic infection), and remaining at higher concentration for the remaining period of the studies. Blood TNF-αconcentrations in both chronically and acutely infected water buffaloes also increased after infection. In the goat experiment, plasma NO concentrations in both infection groups increased from week 3 after infection, and remained higher than that of the control group until the end of the experiment. TNF-α concentrations in goats in infection group 1 and 2 gradually increased after infection and were significantly greater than those of the control group from the 9th to 11th week and from the 11th to 15th week respectively.展开更多
A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and Apri...A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Com- parative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle.展开更多
Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Me...Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Methods:Two experiments were performed, the first one aimed to evaluate the different concentrations (0, 25, 50, 100 μM) of L-ascorbic acid on embryo developmental rate of buffalo oocytes. The L-ascorbic acid was added to the maturation and culture media. In the second experiment, oocytes were cultured in media with two type of antioxidant (ascorbic acid + cysteamine) or ascorbic acid only.Results:There was a significant increase in cleavage rate at 25, 50 μM than 100 μM and control group. But, the blastocyst rate was higher at 50 μM ascorbic acid than other concentrations (0, 25, 100 μM). Supplementation of ascorbic acid and cysteamine to maturation and cultured media improved embryo development than ascorbic acid alone.Conclusions: Using of 50 μM L-ascorbic acid duringin vitro maturation and development improve the developmental competence of buffalo oocytes, this effect was increase with the presence of cysteamine.展开更多
Objective: To clarify the antioxidant effect of butylated hydroxytoulene (BHT) at different concentrations on cooled and post frozen semen diluted in tris-citrate-fructose egg yolk glycerol and lecithin -based extende...Objective: To clarify the antioxidant effect of butylated hydroxytoulene (BHT) at different concentrations on cooled and post frozen semen diluted in tris-citrate-fructose egg yolk glycerol and lecithin -based extenders. Methods: Forty ejaculates were harvested from four buffalo bulls by means of the artificial vagina. Ejaculated semen samples were diluted with each of the tris citrate-fructose egg yolk glycerol and lecithin-based extender diluents. The semen samples diluted with each of the two extenders were added to pre-warmed dried test tubes containing BHT (prepared in ethanol) to get concentrations at 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mM/mL BHT. These ingredients were put at 37 ℃ for 5 min to allow the proper BHT spermatozoal permeation. The diluted semen samples were cooled to 5 ℃ and then frozen to -196 ℃ in 0.25 mL ministraws before dipping in liquid nitrogen pending its evaluation. Sperm motility, viability, morphology, intact acrosome and membrane integrity were tested. Visual motility was tested using a high power ordinary microscope (at 400 × ) with closed circuit television, and sperm concentration was tested using Neubauer haemocytometer and abnormality % using eosin-nigrosin stain. Spermatozoal membrane integrity was tested using the hypo-osmotic swelling test. The sperm with swollen twisting tail was normally intact. Sperm acrosomal integrity % was tested as mentioned by Watson. Results: Addition of BHT improved (P<0.01) progressive motility, viability, morphology and acrosome as well as plasma membrane integrities at 0.5-2.0 mM/mL depending upon types of used extenders and stages of pre-and post-freezing process. Higher levels of 2.5 and 3.0 mM/mL BHT had a deteriorating (P<0.01) result if compared to the control and all extenders assayed. Conclusions: BHT addition at lower concentration can improve pre-frozen and post-thawed buffalo sperm quality.展开更多
Objective:To assess the effects of melatonin and/or cysteamineonin vitro maturation, culturing and post-warming of buffalo embryos.Methods: Buffalo oocytes were classified into control, cysteamine (50 μM), melatonin ...Objective:To assess the effects of melatonin and/or cysteamineonin vitro maturation, culturing and post-warming of buffalo embryos.Methods: Buffalo oocytes were classified into control, cysteamine (50 μM), melatonin (10 ng/mL) and cysteamine (50 μM) + melatonin (10 ng/mL) treatment groups. In experiment 1, previous treatments were added duringin vitromaturation and culturing of buffalo oocytes.Results:Cleavage and blastocyst rates were significantly (P<0.05) increased in melatonin treated group (70.5±0.9 and 12.8±1.0, respectively). However this effect was potentiated when combined with cysteamine (74.0±1.7 and 14.8±1.7, respectively). In experiment 2, the treatements were added in maturtaion, culturing as well as post-warming culture media. Embryos at 7 d were vitrified.Viability assessement directly after warming showed significant increase (P<0.05) in cysteamine, melatonin and their combination groups (76.8±2.8, 80.0±2.1 and 83.3±1.7, respectively) than control (65.8±2.4);but the viability after 24 h post-warming was the best in cysteamine + melatonin combination group (61.4±2.1).Conculsions: Enriching maturation, culturing and post-warming media of buffalo oocytes and embryos with melatonin and/or cysteamine have significantly beneficial effects on oocyte developmental competence as well as embryos vitrification procedure outcomes which in turn resulting in enhancement of commercial buffalo embryo production.展开更多
[ Objectivel The paper aimed to investigate the expression pattern of bbu-miR-103-1 in buffalo (Bubalus bubalis) at lactation and non-lactation periods, and to predict its target gene and function. [ Method] Express...[ Objectivel The paper aimed to investigate the expression pattern of bbu-miR-103-1 in buffalo (Bubalus bubalis) at lactation and non-lactation periods, and to predict its target gene and function. [ Method] Expression pattern of bbu-miR-103-1 at lactation and non-lactation periods were detected by qRT-PCR. The precursor expression plasmid of bbu-miR-103-1 was constructed and named LpEZX-pre-miR-103-1. It was packaged and propagated to produce high-titer lenti- virus in 293T cell lines, which could be used to infect buffalo mammary epithelial cells (BMECs) and over express bbu-miR-103-1. The inhibitor of bbu-miR- 103-1 was chemically synthesized and transfected into BMECs to suppress bbu-miR-103-1 at the same time. The relative expression of pantothenate kinase 3 ( PANK3 ) and milk fat metabolism related genes were detected by qRT-PCR. [ Result] The relative expression of bbu-miR-103-1 at lactation period was 5.29 times higher than that at non-lactation period in buffalo ( P 〈 0.01 ). The LpEZX-pre-miR-103-1 had been successfully constructed and packaged with the infection titer of 3.47×10^6 PFU/mL. Overexpress or suppress of bbu-miR-103-1 extremely down-regulated or up-regulated the expression level of PANK3 in BMECs ( P 〈 0.01 ). Over expression of bbu-miR-103~l extremely enhanced the expression of Acetyl-CoA carboxylase alpha(ACACA), Glycerol-3-phosphate acyhransferase 1 mitochon- drial (GPAM), Diacylglycerol Oacyhransferase l (DGAT1) and Pyrnvate dehydrogenase lipoamide kinase isozyme 4 (PDK4) (P 〈0.01 ), and also significantly up-regulated the expression of sterol regulatory element binding protein-1 c (SREBPI c), Adipose differentiation-related protein (ADFP), Cluster of differentiation 36 ( CD36), Acetyl-CoA synthetase short-chain subfamily member 1 (ACSS1) (P 〈0.05). Over expression of bbu-miR-103-1 down-regulated the expression of PANK3, and improved the mRNA level of SREBPlc by feedback regulation, finally promoting the de novo synthesis of fatty acid beginning with ACACA. [ Conclusion] bbu-miR-103-1 plays an important role in enhancing milk fatty acid synthesis, which provides a molecular base for revealing formation and regulatory mechanism of high-level milk fat in buffalo.展开更多
Objective:To assess the effect of region, season and year of insemination onin-vivo fertility of Italian-Egyptian crossbred buffalo semen.Methods: A total number of 4799 female buffaloes were inseminated by frozen sem...Objective:To assess the effect of region, season and year of insemination onin-vivo fertility of Italian-Egyptian crossbred buffalo semen.Methods: A total number of 4799 female buffaloes were inseminated by frozen semen with at least 50% post-thaw motility of Egyptian-Italian crossbred bulls in three localities in Delta, lower Egypt (El-Behira, El-Sharkia and Damietta) during the period of 2013, 2014 and 2015. The pregnancy rate after two months was evaluated during the four seasons.Results:The rate of pregnancy was significantly (P<0.0001) differ among the three localities. The effect of year of insemination on pregnancy rate was significantly higher during 2014 and 2015 than 2013 in El-Sharkia and El-Behira. But in Damietta, the rate of pregnancy was significantly higher in 2014 than 2013 and 2015. There were no significant differences among seasons in El-Behira and Damietta governorates but there was significant (P<0.05) differences in pregnancy rate in El-Sharkia. It was higher in summer, spring and autumn than in winter.Conclusions:Localities, year of insemination and season of the year have effects on fertility of crossbred Egyptian-Italian buffalo semen.展开更多
基金the Researchers Supporting Project Number(RSP2023 R157),King Saud University,Riyadh,Saudi Arabia.
文摘An illness known as pneumonia causes inflammation in the lungs.Since there is so much information available fromvarious X-ray images,diagnosing pneumonia has typically proven challenging.To improve image quality and speed up the diagnosis of pneumonia,numerous approaches have been devised.To date,several methods have been employed to identify pneumonia.The Convolutional Neural Network(CNN)has achieved outstanding success in identifying and diagnosing diseases in the fields of medicine and radiology.However,these methods are complex,inefficient,and imprecise to analyze a big number of datasets.In this paper,a new hybrid method for the automatic classification and identification of Pneumonia from chest X-ray images is proposed.The proposed method(ABOCNN)utilized theAfrican BuffaloOptimization(ABO)algorithmto enhanceCNNperformance and accuracy.The Weinmed filter is employed for pre-processing to eliminate unwanted noises from chest X-ray images,followed by feature extraction using the Grey Level Co-Occurrence Matrix(GLCM)approach.Relevant features are then selected from the dataset using the ABO algorithm,and ultimately,high-performance deep learning using the CNN approach is introduced for the classification and identification of Pneumonia.Experimental results on various datasets showed that,when contrasted to other approaches,the ABO-CNN outperforms them all for the classification tasks.The proposed method exhibits superior values like 96.95%,88%,86%,and 86%for accuracy,precision,recall,and F1-score,respectively.
文摘A cross-sectional study was conducted to determine the Seropositivity of Toxoplasma gondii in water buffalo in three Iraqi governorates (Baghdad, Dhi Qar, and Maysan) and to estimate the risk aspects related to infestation throughout the period from January to December 2019. A total of 430 serum samples were inspected with a commercial ELISA (Enzyme linked immunosorbent assay) kit. Indirect multi-species kit. The overall Seropositivity of T. gondii in the examined local buffalo was 7.4%, and the highest rate (9.3%) was in Baghdad Governorate. A multivariate regression analysis revealed that adult buffalo (OR = 7.10;95% CI: 0.87-57.68;P = 0.067) and young herds (OR = 8.42;95% CI: 1.07-66.02;P = 0.043) were more subject to infestation from young buffalo and large herds. Furthermore, the hazard of toxoplasmosis was increased in winter especially among animals in contact with cats. It is therefore requisite to determine risk aspects to evaluate which mitigation, control, prevention and procedures should be carried out to diminish, control and prevent infestation with T. gondii and its propagation.
基金funded by the National Natural Science Foundation of China (31972996 and 32160790)Guangxi Bagui Scholar ProgramGuangxi Innovation-Driven Development Project (AA17204051)
文摘Background During mammalian pre-implantation embryonic development(PED),the process of maternal-to-zygote transition(MZT)is well orchestrated by epigenetic modification and gene sequential expression,and it is related to the embryonic genome activation(EGA).During MZT,the embryos are sensitive to the environment and easy to arrest at this stage in vitro.However,the timing and regulation mechanism of EGA in buffaloes remain obscure.Results Buffalo pre-implantation embryos were subjected to trace cell based RNA-seq and whole-genome bisulfite sequencing(WGBS)to draw landscapes of transcription and DNA-methylation.Four typical developmental steps were classified during buffalo PED.Buffalo major EGA was identified at the 16-cell stage by the comprehensive analy-sis of gene expression and DNA methylation dynamics.By weighted gene co-expression network analysis,stage-spe-cific modules were identified during buffalo maternal-to-zygotic transition,and key signaling pathways and biological process events were further revealed.Programmed and continuous activation of these pathways was necessary for success of buffalo EGA.In addition,the hub gene,CDK1,was identified to play a critical role in buffalo EGA.Conclusions Our study provides a landscape of transcription and DNA methylation in buffalo PED and reveals deeply the molecular mechanism of the buffalo EGA and genetic programming during buffalo MZT.It will lay a foundation for improving the in vitro development of buffalo embryos.
文摘Objective:To optimize the concentration of PDC-109 protein in semen for higher cryopreservability and fertility by sequestration or neutralization of PDC-109 by its antibodies(anti-PDC-109 antibodies)in bubaline species.Methods:PDC-109 protein was purified by applying two-step chromatography procedures.Purified protein was injected in rabbits to raise antibodies.These raised anti-PDC-109 antibodies were used in neutralization or sequestration of PDC-109 in in-vitro model.Ejaculates were collected from buffaloes and splited for four groups.Group 1 received egg yolk Tris glycerol extender,without anti-PDC-109 antibodies,while group 2 to 4 received anti-PDC-109 antibodies 266μg/mL,80μg/mL,and 26μg/mL in Tris-fructose-citrate buffer,respectively.Semen quality parameters viz.,forward progressive motility,viability,total morphological abnormality,acrosomal integrity,plasma membrane integrity,cryoinjury and in-vitro zona binding index were evaluated.Results:Semen quality parameters of neat semen were within the normal range of bubaline species.Sperm motility,livability,acrosomal integrity,plasma membrane integrity,and cholesterol content of sperm were decreased and total sperm abnormality was increased significantly in post-thaw semen compared to those in pre-freeze and fresh semen(P<0.05).Semen in group 2 had higher sperm motility,livability,acrosomal integrity,plasma membrane integrity,and cholesterol content of sperm and lower total sperm abnormality significantly compared to those in group 1,3 and 4 at pre-freeze and post-thaw stages(P<0.05).Conclusions:Sequestration or neutralization of PDC-109 by its antibodies significantly improves pre-freeze,and post-thaw semen quality parameters and in-vitro zona binding index with simultaneously reducing cryoinjury or cryodamage in the sperm of bubaline species.
基金This research is funded by the Deanship of Scientific Research at Umm Al-Qura University,Grant Code:22UQU4281755DSR02.
文摘Software needs modifications and requires revisions regularly.Owing to these revisions,retesting software becomes essential to ensure that the enhancements made,have not affected its bug-free functioning.The time and cost incurred in this process,need to be reduced by the method of test case selection and prioritization.It is observed that many nature-inspired techniques are applied in this area.African Buffalo Optimization is one such approach,applied to regression test selection and prioritization.In this paper,the proposed work explains and proves the applicability of the African Buffalo Optimization approach to test case selection and prioritization.The proposed algorithm converges in polynomial time(O(n^(2))).In this paper,the empirical evaluation of applying African Buffalo Optimization for test case prioritization is done on sample data set with multiple iterations.An astounding 62.5%drop in size and a 48.57%drop in the runtime of the original test suite were recorded.The obtained results are compared with Ant Colony Optimization.The comparative analysis indicates that African Buffalo Optimization and Ant Colony Optimization exhibit similar fault detection capabilities(80%),and a reduction in the overall execution time and size of the resultant test suite.The results and analysis,hence,advocate and encourages the use of African Buffalo Optimization in the area of test case selection and prioritization.
文摘In the discipline of Music Information Retrieval(MIR),categorizing musicfiles according to their genre is a difficult process.Music genre classifica-tion is an important multimedia research domain for classification of music data-bases.In the proposed method music genre classification using features obtained from audio data is proposed.The classification is done using features extracted from the audio data of popular online repository namely GTZAN,ISMIR 2004 and Latin Music Dataset(LMD).The features highlight the differences between different musical styles.In the proposed method,feature selection is per-formed using an African Buffalo Optimization(ABO),and the resulting features are employed to classify the audio using Back Propagation Neural Networks(BPNN),Support Vector Machine(SVM),Naïve Bayes,decision tree and kNN classifiers.Performance evaluation reveals that,ABO based feature selection strategy achieves an average accuracy of 82%with mean square error(MSE)of 0.003 when used with neural network classifier.
基金Supported by Science and Technology Program of Yunnan Province(2004NG04)
文摘The Dehong buffalo( D),F1 hybrids of Dehong buffalo × Murrah buffalo( MD) and F1 hybrids of Dehong × Nili buffalo( ND) as the experimental animals to investigate their fatty acid composition and amino acid content. Five of each buffalos were bred by freely grazing and slaughtered at 35. 8-month old age to investigate the fatty acid composition and amino acid content in beef of Dehong buffalo,MD buffalo and ND buffalo. The results showed that,saturated fatty acid content of Dehong buffalo( 44. 33%) was significantly higher than that of ND buffalo( 41. 39%) and MD buffalo( 41. 48%)( P 【 0. 05),and the latter two were not significantly different( P 】 0. 05). The content of PUFA in Dehong buffalo( 53. 90%) was significantly lower than ND( 56. 83%) and MD( 56. 83%)( P 【0. 05),however,the n- 6∶ n- 3( 4. 25) of Dehong buffalo was significantly higher than ND( 3. 14) and MD( 3. 52)( P 【 0. 05). The content of amino acids in muscle was not significantly different between Dehong buffalo,MD buffalo and ND buffalo( P 】 0. 05),while the acidic amino acid content of Dehong buffalo was higher than that of MD buffalo and ND buffalo.
基金Supported by the Innovation Foundation For Postgraduate of Guangxi University(2008105930905D001) the Tackle Key Program in Science and Technology of Science and Technology Bureau of Guangxi Province(0815008-2-4)~~
文摘[Objective] The experiment aimed to set up a method for isolating and culturing endometrial stromal cells (BESC) and endometrial glandular epithelial cells(BEGEC) of buffalo as well as laid foundation for studying biological mechanism of embryo implantation and uterine diseases. [Method] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique were used to isolate BESC and BEGEC, then immunocytochemical method and TRYPAN-Blue assay were used to determine the purity and survival rate of isolated cells. [Result] The BESC and BEGEC were successfully isolated and cultured while immunocytochemical method and cell count method demonstrated that the purity was over 90%. The result of TRYPAN-Blue assay shown that survival rate of BESC and BEGEC was 91% and 78% respectively. [Conclusion] The enzymatic digestion method, scraping method, serial filtration and differential velocity adherent technique could isolate BESC and BEGEC with high purity.
基金Supported by Guangxi Scientific Fund Project (Guikezi0991042, Guikezi 0640015 and Guikezi 0832043)Guangxi Area Education Department Educational and Scientific Layout Project (C, 2006C3)+1 种基金Guangxi Education Department Scientific Research Fund (200709LX075)Guangxi Large Apparatus Collaborated Sharing Net~~
文摘To study the association of oxytocin (OT)'s distribution in hypothalamatic,pituitary and ovary,and understand how the OT secrete releasing in hypothalamus,pituitary and ovaries,the paraffin section immunohistochemistry SuperPicTureTM two step method was used to detect the distribution of OT in hypothalamatic-pituitary-ovary axis of five femal Guangxi local buffalo. The test results could provide morphology according to study the OT's synthesis and mechanism of action,and could play reference and directions part in breeding Guangxi local buffalo. The test results display:oxytocin immuno reactive (OT-IR) neuronsw eremainly distributed arcuate nucleus,supraoptic nucleus and paraventricular nucleus,and OT-IR neurons was also found in ventromedial nucleus,ventrolateralis nucleus,suprachiasmaticus nucleus,dorsomedial nucleus,mamillary body,anterior hypothalamic nucleus and so on. The OT immunoactive production was found in pituitary and few OT-IR nerve fibers extended to post pituitary from hypophyseal stalk and medium eminence. In ovaries,OT immunoactive productions were only distributed in germinal epithelium cells,granulosa cells and lutein cells. The OT was first discovered in singulorum link of hypothalamatic-pituitary-ovary axis of Guangxi local buffalo. The OT immunoactive neurons were first discovered in every main nucleus of Guangxi local buffalo hypothalamus,especially distributed in arcuate nucleus,supraoptic nucleus and paraventricular nucleus.
基金Supported by National High Technology Research and Development (863) Program of China (2011AA100607)National Transgenic Major Project of China (2010ZX08007003)~~
文摘[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of Buffalo. [Method] Buffalo oocytes were randomly divided into two groups 20-22 h after in vitro maturation. One group of oocytes was introduced with about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection 7-10 h or 18-20 h after in vitro fertilization (IVF); the other group of oocytes was introduced with mixture of a single buffalo sperm and about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection (generally called ICSI-Mediated Gene Transfer, ICSI-Tr). Expression of exogenous DNA was observed and recorded during the process of embryonic development. [Result] Early embryonic gene expression efficiency and blastocyst gene expression efficiency in IVF injection group showed no significant difference compared with that in ICSI-Tr group (P0.05). In addition, the cleavage rate and early embryonic gene expression efficiency in IVF injection group were significantly higher with injection at 7-10 h post IVF than that at 18-20 h post IVF (P0.05). [Conclusion] These results indicate that transgenic buffalo embryos can be generated by injecting exogenous DNA into cytoplasm of IVF oocytes, and the optimal injection time is 7-10 h post IVF.
基金supported by the National Key Technology R&D Program of China (2011BAD19B02-6)the Open Grant of Guangxi Provincial Key Laboratory of Water Buffalo Genetics, Breeding and Reproduction, China (SNKF-2012-04)
文摘The aim of this study was to investigate the feasibility of stimulating ovarian fol icle development in order to improve fertility in water buffalo cows by immunization against inhibin. The experiment was carried out in early summer (May) and included 24 multi-parity crossbred Murrah-Swamp buffaloes that were divided into immunized (n=11) and control (n=13) groups. Each immunized cow was administered with a 2-mL immunogen of mineral oil adjuvant containing 2 mg of recombinant inhibinα-subunit fusion protein. The controls were treated with the adjuvant only. Al animals received Ovsynch protocol treatment, starting on the day of the antigen administration, and they were artiifcial y inseminated upon behavioral estrus. As a result, al of the immunized buffaloes generated antibodies against inhibin during the experimental period and had higher plasma concentrations of fol icle-stimulating hormone (FSH), activin, and estradiol (E2) related to estrous expression. A higher proportion of immunized animals expressed estrus behavior than did the controls (72%vs. 30%, P<0.05). On aver-age, inhibin-immunized buffaloes had signiifcantly more large fol icles (≥9 mm in diameter) than the controls (mean±SEM;1.2±0.1 vs. 0.84±0.1, respectively;P<0.05) and a slightly higher mean total number of fol icles (≥2 mm;11.4±0.7 vs. 9.0±1.1, respectively;P=0.09) and smal (2–4 mm) fol icles (8.81±0.6 vs. 6.84±1.0, respectively;P=0.12). A higher percentage of cows ovulated in the immunized group than in the control group (91%(10/11) vs. 54%(7/13), respectively;P<0.05). Moreover, inhibin-immunized cows had slightly larger corpus luteum (CL) than the controls 9 days after ovulation and signiifcantly higher (P<0.01) post-ovulation peak plasma progesterone (P4) concentrations. Immunization against inhibin also mar-ginal y increased the conception rate 42 days after insemination (45.8%vs. 15.4%;P>0.05). These results demonstrate that immunization against inhibin, coupled with the treatment with the Ovsynch protocol, can constitute a new technique to increase fertility in water buffalo cows.
文摘Changes of concentrations were studied in water buffaloes and goats infected with Fasciola he-patica on blood NO(nitric oxide) and TNF-α(tumor necrosis factor-α). Twenty healthy male castrated water buffaloes of 2 - 3 years old and weighing 300 - 500 kg as well as six goats were confirmed free of fasciolosis by fecal examination and Dot-ELISA. Two studies were conducted using the water buffaloes. In the first experiment, 8 water buffaloes were randomly divided into control group (n = 3) and infection group (n = 5). Each buffalo in the infected group received orally 60 metacercariae of F. hepatica per day for 20 days (total 1 200 metacercariae) to produce a chronic infection. In the second experiment, 12 water buffaloes were randomly divided into infected (n=9) and control group (n = 3). Each buffalo in the infected group was given a single oral dose of 1 600 metacercariae to produce an acute infection. The 6 goats were randomly divided into two infected groups and a control group. The sheep in two infections received a single oral dose of 200 and 500 metacercariae respectively, the control group remained uninfected. Blood NO and TNF-a concentrations of the test animals were measured by a reductive enzyme assay and RIA, respectively. Blood NO concentration in both acutely and chronically infected water buffaloes progressively increased from week 3 post-infection and was significantly greater than that of the control group (P<0. 05) at the 5th week (acute infection) and 7th week (chronic infection), and remaining at higher concentration for the remaining period of the studies. Blood TNF-αconcentrations in both chronically and acutely infected water buffaloes also increased after infection. In the goat experiment, plasma NO concentrations in both infection groups increased from week 3 after infection, and remained higher than that of the control group until the end of the experiment. TNF-α concentrations in goats in infection group 1 and 2 gradually increased after infection and were significantly greater than those of the control group from the 9th to 11th week and from the 11th to 15th week respectively.
文摘A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Com- parative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle.
文摘Objective:To improvein vitro embryo production in buffalo by supplementation of L-ascorbic acid during maturation and development (experiment 1) and combination with another antioxidant as cysteamine (experiment 2).Methods:Two experiments were performed, the first one aimed to evaluate the different concentrations (0, 25, 50, 100 μM) of L-ascorbic acid on embryo developmental rate of buffalo oocytes. The L-ascorbic acid was added to the maturation and culture media. In the second experiment, oocytes were cultured in media with two type of antioxidant (ascorbic acid + cysteamine) or ascorbic acid only.Results:There was a significant increase in cleavage rate at 25, 50 μM than 100 μM and control group. But, the blastocyst rate was higher at 50 μM ascorbic acid than other concentrations (0, 25, 100 μM). Supplementation of ascorbic acid and cysteamine to maturation and cultured media improved embryo development than ascorbic acid alone.Conclusions: Using of 50 μM L-ascorbic acid duringin vitro maturation and development improve the developmental competence of buffalo oocytes, this effect was increase with the presence of cysteamine.
文摘Objective: To clarify the antioxidant effect of butylated hydroxytoulene (BHT) at different concentrations on cooled and post frozen semen diluted in tris-citrate-fructose egg yolk glycerol and lecithin -based extenders. Methods: Forty ejaculates were harvested from four buffalo bulls by means of the artificial vagina. Ejaculated semen samples were diluted with each of the tris citrate-fructose egg yolk glycerol and lecithin-based extender diluents. The semen samples diluted with each of the two extenders were added to pre-warmed dried test tubes containing BHT (prepared in ethanol) to get concentrations at 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mM/mL BHT. These ingredients were put at 37 ℃ for 5 min to allow the proper BHT spermatozoal permeation. The diluted semen samples were cooled to 5 ℃ and then frozen to -196 ℃ in 0.25 mL ministraws before dipping in liquid nitrogen pending its evaluation. Sperm motility, viability, morphology, intact acrosome and membrane integrity were tested. Visual motility was tested using a high power ordinary microscope (at 400 × ) with closed circuit television, and sperm concentration was tested using Neubauer haemocytometer and abnormality % using eosin-nigrosin stain. Spermatozoal membrane integrity was tested using the hypo-osmotic swelling test. The sperm with swollen twisting tail was normally intact. Sperm acrosomal integrity % was tested as mentioned by Watson. Results: Addition of BHT improved (P<0.01) progressive motility, viability, morphology and acrosome as well as plasma membrane integrities at 0.5-2.0 mM/mL depending upon types of used extenders and stages of pre-and post-freezing process. Higher levels of 2.5 and 3.0 mM/mL BHT had a deteriorating (P<0.01) result if compared to the control and all extenders assayed. Conclusions: BHT addition at lower concentration can improve pre-frozen and post-thawed buffalo sperm quality.
文摘Objective:To assess the effects of melatonin and/or cysteamineonin vitro maturation, culturing and post-warming of buffalo embryos.Methods: Buffalo oocytes were classified into control, cysteamine (50 μM), melatonin (10 ng/mL) and cysteamine (50 μM) + melatonin (10 ng/mL) treatment groups. In experiment 1, previous treatments were added duringin vitromaturation and culturing of buffalo oocytes.Results:Cleavage and blastocyst rates were significantly (P<0.05) increased in melatonin treated group (70.5±0.9 and 12.8±1.0, respectively). However this effect was potentiated when combined with cysteamine (74.0±1.7 and 14.8±1.7, respectively). In experiment 2, the treatements were added in maturtaion, culturing as well as post-warming culture media. Embryos at 7 d were vitrified.Viability assessement directly after warming showed significant increase (P<0.05) in cysteamine, melatonin and their combination groups (76.8±2.8, 80.0±2.1 and 83.3±1.7, respectively) than control (65.8±2.4);but the viability after 24 h post-warming was the best in cysteamine + melatonin combination group (61.4±2.1).Conculsions: Enriching maturation, culturing and post-warming media of buffalo oocytes and embryos with melatonin and/or cysteamine have significantly beneficial effects on oocyte developmental competence as well as embryos vitrification procedure outcomes which in turn resulting in enhancement of commercial buffalo embryo production.
基金Supported by National Natural Science Foundation of China(31260552)National High-tech Research and Development Plan(863 plan)(2011AA100607)+1 种基金Selection of Excellent Ecological Forage Grass Varieties and Research and Demonstration of Carbon and Nitrogen Source of Fruit-grass Coupling System(GKH 14125008-2-13)Breeding and Popularization of National Approval New Forage Variety Pennisetum purpureum(GYMK 201453057)
文摘[ Objectivel The paper aimed to investigate the expression pattern of bbu-miR-103-1 in buffalo (Bubalus bubalis) at lactation and non-lactation periods, and to predict its target gene and function. [ Method] Expression pattern of bbu-miR-103-1 at lactation and non-lactation periods were detected by qRT-PCR. The precursor expression plasmid of bbu-miR-103-1 was constructed and named LpEZX-pre-miR-103-1. It was packaged and propagated to produce high-titer lenti- virus in 293T cell lines, which could be used to infect buffalo mammary epithelial cells (BMECs) and over express bbu-miR-103-1. The inhibitor of bbu-miR- 103-1 was chemically synthesized and transfected into BMECs to suppress bbu-miR-103-1 at the same time. The relative expression of pantothenate kinase 3 ( PANK3 ) and milk fat metabolism related genes were detected by qRT-PCR. [ Result] The relative expression of bbu-miR-103-1 at lactation period was 5.29 times higher than that at non-lactation period in buffalo ( P 〈 0.01 ). The LpEZX-pre-miR-103-1 had been successfully constructed and packaged with the infection titer of 3.47×10^6 PFU/mL. Overexpress or suppress of bbu-miR-103-1 extremely down-regulated or up-regulated the expression level of PANK3 in BMECs ( P 〈 0.01 ). Over expression of bbu-miR-103~l extremely enhanced the expression of Acetyl-CoA carboxylase alpha(ACACA), Glycerol-3-phosphate acyhransferase 1 mitochon- drial (GPAM), Diacylglycerol Oacyhransferase l (DGAT1) and Pyrnvate dehydrogenase lipoamide kinase isozyme 4 (PDK4) (P 〈0.01 ), and also significantly up-regulated the expression of sterol regulatory element binding protein-1 c (SREBPI c), Adipose differentiation-related protein (ADFP), Cluster of differentiation 36 ( CD36), Acetyl-CoA synthetase short-chain subfamily member 1 (ACSS1) (P 〈0.05). Over expression of bbu-miR-103-1 down-regulated the expression of PANK3, and improved the mRNA level of SREBPlc by feedback regulation, finally promoting the de novo synthesis of fatty acid beginning with ACACA. [ Conclusion] bbu-miR-103-1 plays an important role in enhancing milk fatty acid synthesis, which provides a molecular base for revealing formation and regulatory mechanism of high-level milk fat in buffalo.
文摘Objective:To assess the effect of region, season and year of insemination onin-vivo fertility of Italian-Egyptian crossbred buffalo semen.Methods: A total number of 4799 female buffaloes were inseminated by frozen semen with at least 50% post-thaw motility of Egyptian-Italian crossbred bulls in three localities in Delta, lower Egypt (El-Behira, El-Sharkia and Damietta) during the period of 2013, 2014 and 2015. The pregnancy rate after two months was evaluated during the four seasons.Results:The rate of pregnancy was significantly (P<0.0001) differ among the three localities. The effect of year of insemination on pregnancy rate was significantly higher during 2014 and 2015 than 2013 in El-Sharkia and El-Behira. But in Damietta, the rate of pregnancy was significantly higher in 2014 than 2013 and 2015. There were no significant differences among seasons in El-Behira and Damietta governorates but there was significant (P<0.05) differences in pregnancy rate in El-Sharkia. It was higher in summer, spring and autumn than in winter.Conclusions:Localities, year of insemination and season of the year have effects on fertility of crossbred Egyptian-Italian buffalo semen.