Escherichia coli membrane-derived oxygen-reducing enzyme system (Oxyrase) protects bubaline spermatozoa during cryopreservation.

The objective of this study was to determine the effectiveness of deoxygenation of semen extender using Escherichia coli membrane-derived oxygen scavenger (Oxyrase) on post-thaw quality of buffalo (Bubalus bubalis) spermatozoa. Sixteen semen ejaculates, four each from four bulls, were each divided into five equal fractions, diluted using Tris-egg yolk extender supplemented with different concentrations of Oxyrase (0, 0.3, 0.6, 0.9, and 1.2 U/ml), designated as treatments T1, T2, T3, T4, and T5, respectively, and cryopreserved. Immediately after thawing, Oxyrase did not improve sperm kinetics and motility; however, it improved the keeping quality (significantly lower deterioration of post-thaw sperm motility after incubation for 120 min) in T3. Further, T3 reduced (p < .05) cholesterol efflux and protected the intactness of the sperm plasma membrane. Flow cytometry with Fluo-3 AM/propidium iodide (PI) dual staining revealed the highest (p < .05) proportion of live spermatozoa with low intracellular calcium in T3. Oxyrase supplementation protected spermatozoa from premature capacitation which was confirmed by low expression of tyrosine-phosphorylated proteins (32, 75, and 80 kDa) and a relatively lower percentage of F-pattern (uncapacitated spermatozoa) in chlortetracycline assay. Importantly, the Oxyrase fortification decreased superoxide anion in a dose-dependent manner indicating reduced availability of oxygen at sperm mitochondrial level. Similarly, in Oxyrase-fortified sperm, malondialdehyde concentration, an index of lipid peroxidation, is also reduced in a dose-dependent manner. In conclusion, we demonstrate that deoxygenation of buffalo semen by Oxyrase has the potential of improving post-thaw sperm quality by overcoming the problem of cryocapacitation and oxidative damage during cryopreservation process.

Transabdominal color doppler ultrasonography: A relevant approach for assessment of effects of uterine torsion in buffaloes.

AIM: The present study was conducted on advanced pregnant buffaloes suffering from uterine torsion to assess the status of fetus and uterus by transabdominal ultrasonography, and the findings were compared with normal advanced pregnant buffaloes. MATERIALS AND METHODS: The study was conducted on 20 clinical cases of uterine torsion and 20 normal advanced pregnant buffaloes (control group). The lower ventral area just lateral to linea alba (on both sides of the udder) in standing animals was scanned transabdominally by the two-dimensional convex transducer for various ultrasonographic findings. The data collected were statistically analyzed by "one-way ANOVA" and "independent sample t-test" using computerized SPSS 16.0 software program. RESULTS: Transabdominal ultrasonography revealed dead fetus in 95% uterine torsion cases and proved useful in imaging internal structures of fetuses while no dead fetus was reported in the control group. Size of umbilicus was found significantly decreased (p<0.05) in uterine torsion group in comparison to control animals, but the decrease in placentomal area was marginal (p>0.05) in uterine torsion group. Average thickness of the uterine wall and mean pixel values of fetal fluids (echogenicity) were found significantly increased (p<0.05) in uterine torsion affected buffaloes in comparison to control group. CONCLUSION: Status of fetus (whether live or dead), internal status of uterus, and its contents could be determined by transabdominal ultrasonography in uterine torsion cases and thus determining the prognosis of the uterine torsion cases before going for further manipulations. This will also help in taking all the precautions to avoid death of the fetus.

Two-dimensional and three-dimensional ultrasonography for pregnancy diagnosis and antenatal fetal development in Beetal goats.

AIM: The objective of this study was to compare two-dimensional (2D) and three-dimensional (3D) study of the pregnant uterus and antenatal development of the fetus. MATERIALS AND METHODS: 2D and 3D ultrasound were performed from day 20 to 120 of gestation, twice in week from day 20 to 60 and once in week from day 60 to 120 of gestation on six goats. The ultrasonographic images were obtained using Toshiba, Nemio-XG (Japan) 3D ultrasound machine. RESULTS: On the 20(th) day of gestation, earliest diagnosis of pregnancy was done. First 3D ultrasonographic image of the conceptus, through transabdominal approach, was obtained on day 24. On 39(th) day, clear pictures of conceptus, amniotic membrane, and umbilicus were seen. On 76(th) day of gestation, internal organs of fetus viz heart, kidney, liver, urinary bladder, and stomach were seen both in 2D and 3D images. 3D imaging showed better details of uterine structures and internal organs of the fetus. CONCLUSIONS: Comparing 3D images with 2D images, it is concluded that 2D was better in visualizing fluid while 3D images were better to view details of attachment of fetus with endometrium.