Effect of Physiologically Relevant Heat Shock on Development,Apoptosis and Expression of Some Genes in Buffalo (Bubalus bubalis) Embryos Produced In Vitro

For investigating the effects of physiologically relevant heat shock, buffalo oocytes/embryos were cultured at 38.5°C (control) or were exposed to 39.5°C (Group II) or 40.5°C (Group III) for 2 h once every day throughout in vitro maturation (IVM), fertilization (IVF) and culture (IVC). Percentage of oocytes that developed to 8‐cell, 16‐cell or blastocyst stage was lower (p < 0.05) and the number of apoptotic nuclei was higher (p < 0.05) for Group III > Group II > controls. At both 8–16‐cell and blastocyst stages, relative mRNA abundance of stress‐related genes HSP 70.1 and HSP 70.2 and pro‐apoptotic genes CASPASE‐3, BID and BAX was higher (p < 0.05) in Groups III and II than that in controls with the exception of stress‐related gene HSF1. Expression level of anti‐apoptotic genes BCL‐XL and MCL‐1 was also higher (p < 0.05) in Groups III and II than that in controls at both 8–16‐cell and blastocyst stages. Among the genes related to embryonic development, at 8–16‐cell stage, the expression level of GDF9 was higher (p < 0.05) in Group III than that in controls, whereas that of GLUT1, ZAR1 and BMP15 was not significantly different among the three groups. At the blastocyst stage, relative mRNA abundance of GLUT1 and GDF9 was higher (p < 0.05) in Group II than that in controls, whereas that of ZAR‐1 and BMP15 was not affected. The results of this study demonstrate that exposure of buffalo oocytes and embryos to elevated temperatures for duration of time that is physiologically relevant severely compromises their developmental competence, increases apoptosis and affects stress‐, apoptosis‐ and development‐related genes.     

Buffalo (Bubalus bubalis) ES Cell–Like Cells are Capable of In Vitro Skeletal Myogenic Differentiation

When buffalo embryonic stem (ES) cell–like cells that expressed surface markers SSEA‐4, TRA‐1‐60, TRA‐1‐81, CD9 and CD90 and intracellular markers OCT4, SOX2 and FOXD3, as shown by immunofluorescence, and that expressed REX‐1 and NUCLEOSTEMIN as confirmed by RT‐PCR, were subjected to suspension culture in hanging drops in absence of LIF and buffalo foetal fibroblast feeder layer support, they differentiated to form three‐dimensional embryoid bodies (EBs). Of 231 EBs examined on Day 3 of suspension culture, 141 (61.3 ± 3.09%) were of compact type, whereas 90 (38.4 ± 3.12%) were of cystic type. The cells obtained from EBs were found to express NF‐68 and NESTIN (ectodermal lineage), BMP‐4 and α‐skeletal actin (mesodermal lineage), and α‐fetoprotein, GATA‐4 and HNF‐4 (endodermal lineage). When these EBs were cultured on gelatin‐coated dishes, they spontaneously differentiated to several cell types such as epithelial‐ and neuron‐like cells. When EBs were cultured in the presence of 1 or 2% DMSO or 10^?8 m or 10^?7 m retinoic acid for 25 days, ES cells could be directed to form muscle cell–like cells, the identity of which was confirmed by expression of α‐actinin by immunofluorescence and of MYF‐5, MYOD and MYOGENIN genes by RT‐PCR. MYOD was first detected on Day 10 in both treatment groups and on Day 15 in controls, whereas MYOGENIN was first detected on Day 10, Day 15 and Day 25 in the presence of retinoic acid, in the presence of DMSO and in controls, respectively. The present study demonstrates the ability of buffalo ES cell–like cells to undergo directed differentiation to cells of skeletal myogenic lineage.     

Radiographic assessment of pulmonary fluid clearance and lung aeration in newborn calves delivered by elective Caesarean section

The aim of this study was to develop a radiographic standard for the assessment of pulmonary fluid clearance and lung aeration in newborn calves. Caesarean‐delivered mature calves (n = 9) underwent lung assessment by thoracic radiography as well as arterial and venous blood gas analysis within the first 30 min, 1, 2, 3, 6, 12 and 24 hr after birth. The results indicated that newborn calves delivered by elective Caesarean section suffered from a physiological combined respiratory and metabolic acidosis with the dominance of respiratory acidosis, and an improvement in these conditions was recorded within 24 hr after birth. Concerning the radiographic results, clear lung fields, improvement in lung expansion, air content of the lung and absence of lung opacification occurred within 24 hr of birth. Furthermore, the ventral lung quadrant showed an improvement in radiographic opacification and lung expansion earlier than the dorsal lung regions. The findings of this study support the potential role of thoracic radiography in the assessment of pulmonary fluid clearance and lung aeration in newborn calves.     

Classification of soils supporting mangrove plantation in Kuwait

The State of Kuwait has initiated an ambitious national program to establish greenery in 20000 ha of open spaces in inland and coastal areas. Afforestation of intertidal zones with mangrove plants is being considered as a viable option for greening coastal areas. In view of this fact, experimental plantations were established using two ecotypes of Avicennia marina (Bahrain and UAE) and one ecotype of Avicennia germinans (Florida). The study consisted of two 10-year old plantations and two 2-year old plantations. Seedlings established successfully and grew vigorously on some of the locations within the site. Therefore, studies were conducted during 2002-2003 to describe and classify soils that supported maximum mangrove establishment and growth under the harsh arid coastal conditions of Kuwait. For this purpose, eight soil profiles (four each representing areas where seedlings established successfully and grew vigorously and where all seedlings died in the initial stages of establishment) were investigated and classified according to the soil taxonomy. The main soil types observed in the study site were: Typic Aquisalids, Typic/sodic Aquicambids and Calcic Aquisalids. Mangrove growth performance in relation to the landform, soil type and properties of the soil at various depths is discussed in the paper. The plant growth appeared to be the better on the typic Aquisalid soil than that on the Typic/sodic Aquicambid soils. The relative proportions of the coarse sand fraction, occurrence of the anaerobic layer in the profile and the surface layer salinity influenced both the establishment and growth of mangrove plants.     

Effect of Artificial Insemination Protocol and Dose of Frozen/Thawed Stallion Semen on Pregnancy Results in Mares

Deep intra‐uterine insemination is commonly accepted as a routine procedure for artificial insemination in horses. The motives and principles of deep insemination are well described, but the equipment used may differ. In this trial, the efficiency of two different insemination pipettes for deep intra‐uterine insemination in the mare was compared with insemination into the uterine body using commercially available frozen–thawed semen of two stallions of proven fertility. These inseminations were performed using two different doses. The semi‐flexible Minitube pipette was compared with a newly designed insemination device with a more flexible telescopic insemination catheter (Ghent device). The semi‐flexible Minitube pipette performed better than the newly designed insemination device with respect to pregnancy outcome (p = 0.008). The superiority of deep horn insemination over uterine body insemination was reflected by the better pregnancy rates obtained after deep insemination using the same low doses (30.6% better pregnancy rates) (p = 0.0123).     

Testing newly introduced ornamental plants to the arid climate of Kuwait

Thirty five ornamental plants were introduced into the Ahmadi Bioremediated Soil Park to screen and closely monitor the growth performance of these new introductions under the local environmental conditions. Out of these 35 plant species, 15 were planted in both bioremediated and agricultural soils to assess their suitability for growing in bioremediated soils. Data on vegetative growth was recorded to assess the performance of newly-introduced plants, whereas, the heat stress was documented of Bauhinia blakeana, where no visible effects of petroleum pollutants on plant growth were observed. Performance of each plant group is discussed separately in the text. Overall, plant survival was 72.73% for all plant groups. Generally, plants that failed to grow in bioremediated soil also did not survive in the agricultural soil, indicating that they were sensitive to heat stress more than the presence of petroleum contaminants in the soil. New plant species that were able to establish and perform satisfactorily will be used in greenery projects after they are fully naturalized to the prevailing climatic conditions.