Effect of Taurine and Melatonin in the Culture Medium on Buffalo In Vitro Embryo Development

This study was carried out to investigate the effect of supplementing culture medium with different concentrations of taurine and melatonin, on buffalo oocyte in vitro meiotic maturation and embryo development. In experiment 1, oocytes were matured in vitro and the cleaved embryos were cultured in the same following seven culture medium; (i) control (TCM 199 + 10% SS); (ii) control + 0.5 m m taurine; (iii) control + 1 m m taurine; (iv) control + 3 m m taurine; (v) control + 5 μ m melatonin; (vi) control + 10 μ m melatonin and (vii) control + 50 μ m melatonin. In experiment 2, based on the results of experiment 1, to examine the synergistic effect of antioxidants, the oocytes were matured in culture medium (TCM199 + 10% SS), supplemented with both taurine at 1 m m and melatonin at 10 μ m concentration and the cleaved embryos were cultured in the same medium. Supplementation of taurine at 1 m m concentration in the culture medium resulted in a higher (p < 0.05) transferable embryo (TE) yield when compared with control (20.6% vs 14.1%). Supplementation of melatonin at 10 and 50 μ m concentration in the culture medium resulted in a higher (p < 0.05) meiotic maturation rate (90.3% and 88.8% respectively) and TE yield (28.4% and 27.2% respectively), than the other treatments. In experiment 2, the TE yield did not improve by supplementing the culture medium with both taurine and melatonin, when compared with melatonin alone. In conclusion, the results of this study demonstrated that, enriching the culture medium with taurine and melatonin, improves in vitro embryo production efficiency in buffaloes. In particular, a high TE yield was obtained by enriching the culture medium with 10 μ m melatonin.     

Difference in virulence between Staphylococcus aureus isolates causing gangrenous mastitis versus subclinical mastitis in a dairy sheep flock

Staphylococcus aureus mastitis in dairy sheep ranges from subclinical mastitis to lethal gangrenous mastitis. Neither the S. aureus virulence factors nor the host-factors or the epidemiological events contributing to the different outcomes are known. In a field study in a dairy sheep farm over 21 months, 16 natural isolates of S. aureus were collected from six subclinical mastitis cases, one lethal gangrenous mastitis case, nasal carriage from eight ewes and one isolate from ambient air in the milking room. A genomic comparison of two strains, one responsible for subclinical mastitis and one for lethal gangrenous mastitis, was performed using multi-strain DNA microarrays. Multiple typing techniques (pulsed-field-gel-electrophoresis, multiple-locus variable-number, single-nucleotide polymorphisms, randomly amplified polymorphic DNA, spa typing and sas typing) were used to characterise the remaining isolates and to follow the persistence of the gangrenous isolate in ewes’ nares. Our results showed that the two strains were genetically closely related and they shared 3 615 identical predicted open reading frames. However, the gangrenous mastitis isolate carried variant versions of several genes (sdrD, clfA-B, sasA, sasB, sasD, sasI and splE) and was missing fibrinogen binding protein B (fnbB) and a prophage. The typing results showed that this gangrenous strain emerged after the initial subclinical mastitis screening, but then persisted in the flock in the nares of four ewes. Although we cannot dismiss the role of host susceptibility in the clinical events in this flock, our data support the hypothesis that S. aureus populations had evolved in the sheep flock and that S. aureus genetic variations could have contributed to enhanced virulence.     

Scavenging of free radicals, antimicrobial, and cytotoxic activities of the Maillard reaction products of beta-lactoglobulin glycated with several sugars

The Maillard reaction occurs during many industrial and domestic thermal treatments of foods. It is widely used because of its role in creating colors, flavors, textures, and other functional properties in foodstuffs. Proteins glycated without the use of conventional chemical reagents have improved technofunctional properties such as heat stability, emulsifying, and foaming properties. The present study was carried out to determine the extent to which this reaction can convey antioxidant, antimicrobial, or cytotoxic activities to beta-lactoglobulin (BLG) and to its tryptic and peptic hydrolysates. BLG was modified with six different sugars in solution at 60 degrees C. Antiradical properties were estimated using a radical scavenging activity test. Antimicrobial activities against different bacterial strains were studied with a diffusion disk method. Cytotoxic tests were performed using two cell lines and the 3-(4,5-dimethylthiazoyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) rapid colorimetric assay. Glycation induced a radical scavenging activity to BLG, the intensity of which depended on the sugar used for modification. Proteins modified with ribose and arabinose showed the highest radical scavenging activities depicted by about 80 and 60% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) absorption decrease at 515 nm. No antimicrobial effect of any glycated form of BLG against Escherichia coli, Bacillus subtilis, Listeria innocua, and Streptococcus mutans was observed. The MTT test showed no enhancement of cytotoxicity by modified proteins and peptides against COS-7 and HL-60 cells. Thus, glycated proteins could be used in formulated food as functional ingredients with a radical scavenging activity able to delay deterioration due to oxidation. This use could be even more advisable considering the lack of toxicity to eukaryotic and prokaryotic cell cultures demonstrated in this work.     

Kinetic study of the acid hydrolysis of various oligofructose samples

The kinetic of acid hydrolysis of five commercially available oligofructose samples used as food ingredients has been investigated as a function of the dry matter concentration, reaction pH, and temperature. The initial fructose release rate is found to be roughly proportional to the inverse of the average polymerization degree in number. A pseudo first order kinetic is found with respect to the fructosyl chain end concentration and to the proton concentration. Arrhenius plot is found to reasonably fit the data in a relatively wide temperature range (7-130 degrees C). The results allow the estimation of the fructose release rate in many foodstuff processing conditions.     

Detection of Follicular Apoptosis in Water Buffalo (Bubalus bubalis) Ovary by Histology and Nick End Labelling Technique

The objective of this experiment was to assess the features and extent of follicular apoptosis in the water buffalo (Bubalus bubalis) ovary using classical histology and nick end labelling technique. Ovaries (n = 40) procured from the slaughterhouse were used for the study. The sections (5 μm) were used for detection of terminal deoxynucleotidyl transferase‐mediated dUTP‐biotin nick end labelling (TUNEL) and classical histology (H&E). Those follicles showing ≥ 5% TUNEL positivity (TUNEL assay) and pyknotic nuclei (histology) in granulosa cells were classified as atretic. Based on histology, the atretic primary and secondary follicles (%) were 93.82 and 95.62 respectively. The histology study reveals that the rates (%) of atresia in <1, 1–3, 3–5 mm and >5 mm were 36.90, 40.50, 62.84 and 74.5 respectively. Further the atretic tertiary follicles (%) were significantly lower than the primary and secondary classes of follicles. TUNEL assay reveals that the atretic rate (%) of tertiary follicles in <1, 1–3, 3–5 and ≥ 5 mm class follicles were 50.88, 53.84, 81.81 and 36.36 respectively. The percentage of atresia in >5 mm diameter follicles were significantly lower in TUNEL than histology. Percentages of granulosa and thecal cells positive for atresia by TUNEL were 30.7 ± 0.53 and 13.82 ± 0.18 respectively per follicle. The initial structural changes in atretic follicles were seen primarily in the granulosa cells. In severely atretic follicles TUNEL positive granulosa cells along with theca cells have to be considered in assessing the rate and extent of atresia.     

Slip-rate measurements on the Karakorum Fault may imply secular variations in fault motion

Beryllium-10 surface exposure dating of offset moraines on one branch of the Karakorum Fault west of the Gar basin yields a long-term (140- to 20-thousand-year) right-lateral slip rate of approximately 10.7 +/- 0.7 millimeters per year. This rate is 10 times larger than that inferred from recent InSAR analyses ( approximately 1 +/- 3 millimeters per year) that span approximately 8 years and sample all branches of the fault. The difference in slip-rate determinations suggests that large rate fluctuations may exist over centennial or millennial time scales. Such fluctuations would be consistent with mechanical coupling between the seismogenic, brittle-creep, and ductile shear sections of faults that reach deep into the crust.     

Detection of circumstellar material in a normal type Ia supernova

Type Ia supernovae are important cosmological distance indicators. Each of these bright supernovae supposedly results from the thermonuclear explosion of a white dwarf star that, after accreting material from a companion star, exceeds some mass limit, but the true nature of the progenitor star system remains controversial. Here we report the spectroscopic detection of circumstellar material in a normal type Ia supernova explosion. The expansion velocities, densities, and dimensions of the circumstellar envelope indicate that this material was ejected from the progenitor system. In particular, the relatively low expansion velocities suggest that the white dwarf was accreting material from a companion star that was in the red-giant phase at the time of the explosion.     

Molecular Basis of Gene-for-Gene Specificity in Bacterial Speck Disease of Tomato

Transient expression of the Pseudomonas syringae avirulence gene avrPto in plant cells resulted in a Pto-dependent necrosis. The AvrPto avirulence protein was observed to interact directly with the Pto resistance protein in the yeast two-hybrid system. Mutations in the Pto and avrPto genes which reduce in vivo activity had parallel effects on association in the two-hybrid assay. These data suggest that during infection the pathogen delivers AvrPto into the plant host cell and that resistance is specified by direct interaction of Pto with AvrPto.     

Nutrient supply enhances both IGF-I and MSTN mRNA levels in chicken skeletal muscle

Nutrient supply may control muscle growth directly and indirectly through its influence on regulatory factors. The present study focuses on its effects on muscle insulin-like growth factors (IGF-I and -II) and myostatin (MSTN). Their mRNA levels were quantified by real time RT-PCR in pectoralis major (PM) and sartorius (SART) muscles from broiler chickens submitted to different feeding regimens (fed or fasted for 48 h) between hatch and 2 days of age and at 4 weeks of age. In the PM of 4 weeks old broilers, mRNA levels were also evaluated after a 16 h-fast and a refeeding period (refed 24 or 48 h after a 48 h-fast). In the PM muscle, both IGF-I and MSTN mRNA levels increased between 0 and 2 days of age in the fed group, while they remained low in the unfed one. A comparable trend was observed in the SART, but with lesser amplitude. In both muscles of 4 weeks old chickens, a 48 h-fast induced a significant reduction in MSTN mRNA levels (20% of fed state). In the PM, this effect required more than 16 h of fasting to occur and was fully reversed by only 24h of refeeding. IGF-I mRNA levels also varied with nutritional state. They decreased significantly with fasting in the SART muscle. By contrast, IGF-II mRNA levels did not vary significantly. Our data shows for the first time that two major paracrine regulators of muscle growth, IGF-I and MSTN, are sensitive to nutrient supply in hatching chicks, and also that fasting reduced IGF-I and MSTN mRNA levels in muscles of older chickens.