Production of monoclonal antibody against recombinant bovine sex-determining region Y (SRY) and their preferential binding to Y chromosome-bearing sperm

Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms.     

Effects of ammonia on growth and molting of Litopenaeus vannamei postlarvae reared under two salinity levels

ABSTRACT

Litopenaeus vannamei postlarvae were exposed to 0, 6, 13, and 19 mg/L total ammonia nitrogen (TAN) treatments. After 45 days, shrimp weight and length were lowest under TAN concentrations of 13 and 19 mg/L (P ≤ 0.05). Maximum weight gain was observed in control and 6 mg/L treatments. Mortality was highest (80.55 ± 4.80%) under 19 mg/L reared in 35 ppt salinity. Average intermolt periods of PLs exposed to 0, 6, 13, and 19 mg/L TAN were 11.5 ± 0.7, 10.8 ± 1.3, 9.4 ± 1.0, and 8.7 ± 0.6 days under 35 ppt and 11.1 ± 0.5, 10.7 ± 0.6, 10.1 ± 0.5, and 9.5 ± 0.2 days under 45 ppt salinity. Although TAN increased postlarvae molting frequency, its negative effects on the shrimp growth and survival of PLs was directly linked to its concentration and exposure duration. Higher salinity reduces the effects of ammonia and increases the survival.     

Immobilization of Urease onto Modified Egg Shell Membrane through Cross Linking

Background:Immobilization is an approach in industry to improve stability and reusability of urease. The efficiency of this technique depends on the type of membrane and the method of stabilization. Methods:The PEI-modified egg shell membrane was used to immobilize urease by absorption and glutaraldehyde cross-linking methods. The membranes were characterized by FTIR and AFM, and Nessler method was applied to measure the kinetic of the immobilized enzymes. Finally, the storage stability (6 °C for 21 days) and reusability (until enzyme activity reached to zero) of the immobilized enzymes were investigated. Results:Based on FTIR, three new peaks were observed in both the absorption- (at 1389.7, 1230.8, and 1074.2 cm^-1) and the cross-linking (at 1615-1690, 1392.7, 1450 cm^-1) immobilized enzymes. The surface roughness of the native membrane was altered after PEI treatment and enzyme immobilization. The optimal pH of cross-linking immobilized enzymes was shifted to a more neutral pH, while it was alkaline in adsorption-immobilized and free enzymes. The reaction time decreased in all immobilized enzymes (100 min for free enzyme vs. 60 and 30 min after immobilizing by adsorption and cross-linking methods, respectively). The optimal temperature for all enzymes was 70 °C and they had a higher K_m and a lower V_max than free enzyme. The stability and reusability of urease were improved by both methods. Conclusion:Our findings propose these approaches as promising ways to enhance the urease efficiency for its applications in industries and medicines. Key Words: Egg shell, Immobilization, Polyethylenimine, Urease     

Compost Amendment to Sandy Soil Affects Soil Properties and Greenhouse Tomato Productivity

Sandy soils, with low productivity, could be improved by compost application to sustain crop production. This study aimed to examine the effect of three compost types (vegetable, fruit and yard waste compost, garden waste compost, and spent mushroom compost) on basic properties of a loamy sand and greenhouse tomato productivity. Disturbed and intact soil samples were taken from a decade-long compost field experiment on loamy sand with three compost types at application rate of 30 m³ ha^?1 yr^?1 (7.5 ton ha^?1 yr^?1). The soils were characterized for chemical and physical properties. Tomato was planted in a greenhouse using soil samples from the field and vegetative and yield parameters (plant height, stem diameter, leaf number, and fruit yield), water productivity, and harvest index were evaluated. All compost types significantly increased soil total carbon, total nitrogen, pH, electrical conductivity and significantly decreased bulk density, with no effect on plant available water compared to the control. Fresh and dry fruit weights were significantly increased after compost addition. Plant height, leaf number, stem diameter, and total biomass did not significantly improve after compost addition. Spent mushroom compost had greater effect in improving tomato productivity. A decade-long application of composts on loamy sand improved basic chemical and physical properties which were reflected in increased fruit yield in tomato. Since no negative effect of compost was observed, we suggest that sandy soils may serve as a safe end use option for these composts and potentially support crop growth.     

Micropropagation of Prunus scoparia,a Suitable Rootstock for Almond under Drought Conditions

Prunus scoparia is a wild deciduous shrub, usually living on dry calcareous soils of the rocky mountains and has been used as a grafting rootstock for domesticated almonds to provide drought resistance. In the current study, micropropagation ability of P. scoparia was investigated using cytokinin and auxin. Uniform nodal shoot pieces (3–5 cm in length) of seedlings were used as explants. The explants were disinfected with 10% sodium hypochlorite solution. For adventitious shoot induction and proliferation, Murashige and Skoog (MS) media containing 7.00 g/l agar and 30.00 g/l sucrose containing five concentrations of benzyl adenine (BA) (0.00, 0.50, 1.00, 2.00, and 4.00 mg/1) and also containing six concentrations of Thidiazuron (TDZ) (0.00, 0.50, 1.00, 2.00, 5.00, and 7.00 mg/1) were compared. For rooting, in vitro shoots (2–3 cm) were transferred into ½ MS medium supplemented with 30 g/l sucrose, 7.50 g/l agar, and different concentrations of IBA (0.00, 0.25, 0.50, and 1.00 mg/l) and NAA (0.00, 0.25, 0.50, and 1.00 mg/l). Based on the results obtained for shoot proliferation, only 2.00 and 4.00 mg/l BA and 2.00 mg/l TDZ concentrations generated shoots, while other treatments did not show shoot proliferation. Among the three treatments that generated shoots, the best results for shoot number, leaf number, and leaf color quality were observed in media containing 2.00 mg/l TDZ. Based on the results obtained for rooting, the effect of IBA concentrations on the rooting percentage, root number, and root length was significant. Among IBA concentrations, only 0.50 mg/l IBA induced rooting, while there was no rooting in the media containing other IBA concentrations. None of the NAA concentrations showed rooting. In conclusion, MS culture medium supplemented with 2.00 mg/l TDZ and ½ MS culture medium supplemented with 0.50 mg/l IBA are suggested for in vitro shoot proliferation and rooting of P. scoparia, respectively. The results presented herein could be used for in vitro selection and micropropagation of P. scoparia.     

Beneficial effects of alternative lighting schedules on the incidence of ascites and on metabolic parameters of broiler chickens

The beneficial effects of different lighting programmes on the incidence of ascites was investigated in an experiment with 360 three-day-old male broiler chickens. At 3 days of age, chicks were randomly divided over three rooms in a high-altitude farm, 2000 m above sea level. During days 14 to 28 ambient temperature decreased during the night but the minimum temperature did not descend below 15 degrees C. In the first room the continuous lighting schedule (CL, 23L:1D) was maintained and in the second room an intermittent lighting schedule (IL, 1L:3D), repeated six times daily, was imposed from 3 days of age. In the third room, an increasing photoperiod schedule (IP, 4 to 14 days, 6L:18D; 15 to 21 days, 10L:14D; 22 to 28 days, 14L:10D; 29 to 35 days, 18L:6D; 36 to 42 days, 23L:1D) was provided. Mortality associated with right ventricular failure and ascites was numerically lower in birds reared under the IL and IP schedules compared to birds reared under the CL schedule, which can be attributed to the temporary reduction in relative growth and feed intake in IL and IP birds. It was concluded that the beneficial effect of lighting schedules could be due to a reduced metabolic rate as a consequence of the altered growth trajectory, as also reflected in the lower haematocrit and plasma T3 levels of IL and IP birds compared to CL birds.     

Opioid receptors of the central amygdala and morphine-induced antinociception

BACKGROUND: The amygdala is a forebrain region, which is known as a modulator of pain sensation. The amygdala, particularly the central nucleus, has high concentrations of enkephalins relative to dynorphins and has high concentrations of opioid receptors. We here studied the role of central nuclei of amygdala in morphine antinociception. METHODS: In this study, we used 130 male Wistar rats (200-250 g). Bilateral two guide cannula were inserted into central nuclei of amygdala. The drugs were administrated via intra central-amygdala and intraperitoneal. The antinociceptive effect was measured by formalin test. RESULTS: Bilateral microinjections of morphine (50 and 100 microg/rat) into the central nuclei of amygdala elicited powerful suppression of nociceptive behaviors in both phases of formalin test. The intraperitoneal administration of naloxone (1 and 2 mg/kg) decreased significantly the antinociception induced by the intra-amygdaloid injection of morphine. Our data also showed that microinjection of naloxone (50 and 100 microg/rat) into the central nuclei of amygdala could reduce the analgesic effects of systemic morphine (7 mg/kg). On the other hand, bilateral neurotoxic lesions of the central nuclei of amygdala attenuated the antinociception induced by subcutaneous or intra-amygdaloid injection of morphine. CONCLUSION: These findings suggest that morphine analgesia in the formalin test depends on ascending connections to the forebrain, probably the amygdala.     

Willingness to travel to avoid recreation conflicts in Danish forests

Conflicts among forest visitors have direct effects on the quality of a recreational experience. As the number of visitors to forests close to residential areas increases, as well as the number of different activities, so does the potential for perceived conflicts. According to the literature, expanding knowledge of conflict characteristics and their causes is important for recreation planners and managers who aim to reduce conflicts.In the present study, different forest user groups were identified and categorised according to their pursued activities, and for each group, causes of conflict were identified. Furthermore, a choice experiment was constructed to estimate the distance visitors are willing to travel to encounter few visitors as opposed to many visitors, and thereby potentially experience fewer conflicts. Comparing the marginal willingness to travel (WTT) of different user groups suggests that some groups have a WTT further than the average to reach a forest with ‘Few’ visitors. The average WTT to reach a forest area with ‘Few’ visitors. ‘Mountain bikers,’ ‘Peace and nature lovers’ and ‘Horse riders’ are willing to travel 4 km more than the average per visit to reach a less crowded forest. At the other end of the scale, we find that people who are doing physical exercise are willing to travel 2 km less than the average to reach a less crowded forest.     

Differentiation of human umbilical cord mesenchymal stem cell into germ-like cell under effect of co-culture with testicular cell tissue

Supplements produced by mouse testicular cells (mTCs) and the interaction between cells can increase the differentiation rate of human umbilical cord mesenchymal stem cells (hUCMSCs) into the germ-like cells. We studied the differentiation rate of hUCMSCs into the germ-like cells under effect of mTCs co-culturing. Isolated hUCMSCs from postpartum human umbilical cords were cultured. Then, the expression of mesenchymal (CD73, CD90 and CD105) and haematopoietic (CD34 and CD45) markers of hUCMSCs were confirmed by flow cytometry. Then, the hUCMSCs were cultured in four distinct groups: (a) control, (b) co-culture until D0, (c) co-culture until D5 and (d) co-culture until D10, in order to differentiate into the germ-like cells. After 10 days, the expression of OCT4, VASA, Fragilis and SYCP3 genes were examined by Real-Time qPCR. The flow cytometry indicated a high expression of mesenchymal markers and a low expression of haematopoietic markers (CD73:98.6%, CD90: 99.1%, CD105: 99.5%, CD34: 4.22% and CD45: 2.54%). The expression of OCT4 decreased during the time while the expression of VASA, Fragilis and SYCP3 markers increased in the co-culture with testicular cells (p value <.05). Co-culture with mTCs may be used as an effective method to differentiate hUCMSCs into germ-like cells.