Developmental competence of in vitro matured ovine oocytes vitrified in solutions with different concentrations of trehalose

This study aimed to determine the optimum concentration of trehalose in solutions used for vitrification of in vitro matured (IVM) ovine oocytes. IVM oocytes were randomly divided into four experimental (vitrified) and one control (fresh) groups. Experimental groups were treated with different concentrations (0.0, 0.25, 0.5 and 1.0 M) of trehalose. After warming, some viable oocytes were exposed to 0.25% pronase to test zona pellucida hardening, whereas the others were fertilized and cultured in vitro for 8 days to evaluate their developmental competence. Blastocysts quality was assessed by differential staining and TUNEL test. Survival and developmental rates of oocytes vitrified in the presence of 0.5 M trehalose were significantly higher than those of the other vitrified groups. Furthermore, there was a significant difference between fresh and vitrified groups in total blastocyst rate. Analysis of blastocysts quality also revealed a significant difference between the group treated with 0.5 M trehalose and other groups in terms of apoptotic index. Furthermore,zona pellucida digestion time period was longer in trehalose‐free (0.0 M) group compared to other groups. In conclusion, we found that IVM ovine oocytes vitrified in solutions containing 0.5 M trehalose are fertilization‐competent and are able to produce good‐quality blastocysts with an apoptotic index comparable to that of the fresh oocytes. Therefore, 0.5 M may be considered the optimum concentration of trehalose to be used in solutions prepared for vitrification of oocytes.     

Vitrification of mouse preantral follicles versus slow freezing: Morphological and apoptosis evaluation

The aim of this study was evaluation of survivability, maturation rate and apoptotic gene expression of preantral follicles after vitrification and slow freezing technique. Normal mouse preantral follicles were randomly divided into three experimental groups. In the control group, follicles were cultured immediately; in the vitrification and slow freezing groups, follicles were cultured after vitrification‐warming and slow freezing‐thawing procedures. Follicular viability was assessed by using 0.4% trypan blue, and molecular evaluation of messenger RNA levels of apoptosis‐related genes was performed by the semi‐quantitative RT‐PCR method after 3 h of culture. Oocyte maturation rates were also evaluated on day 14 of culture. Survival and maturation rate in the slow freezing group were significantly lower than those in control and vitrification groups (P ≤ 0.05). Although there was no difference in Survivin expression among the three experimental groups, Bcl‐2 expression was significantly lower in the slow freezing group compared to the other groups (P ≤ 0.05). The expression of Bax, P53, Fas and Bax/Bcl‐2 ratio in the slow freezing group was significantly higher than control and vitrification groups (P ≤ 0.05). Preantral follicle vitrification seems to be better than slow freezing as seen in the survival, maturation and expression rates of apoptotic gene variants.     

Microbial and sensory characteristics of camel meat during refrigerated storage as affected by gamma irradiation

The present study was undertaken to assess the microbiological profile of fresh camel meat and the possibility of improving microbial quality and extending the refrigerated storage life of meat by using low-dose gamma irradiation. Camel meat samples were subjected to 0 (control), 1.5 and 3 kGy doses and stored at 3 +/- 1 degrees C. the microbial and sensory attributes were evaluated. Exposure to 1.5 kGy dose significantly reduced the initial counts of Aerobic Plate Counts (APCs), psychrophilic bacteria, Lactic Acid Bacteria (LAB), molds and yeasts, Staphylococcus aureus, Listeria monocytogenes and Enterococci. Moreover, Pseudomonas, coliforms and Escherichia coli were below the detection levels. Irradiation at 3 kGy significantly reduced the initial counts of APCs LAB and Enterococci by 99.5, 93.5 and 93.9%, respectively. Pseudomonas, coliforms, S. aureus, L. monocytogenes and E. coli were not found at dose of 3 kGy during entire storage period, also psychrophilic bacteria and molds and yeasts were below the detection levels during 6 days of storage. This study shows that irradiation had no significant effects on the sensory attributes of camel meat. Refrigerated shelf-life of the meat irradiated at 1.5 and 3 kGy were 15 and 21 days, respectively, compared to 7 days for non-irradiated controls.     

Evaluation of different media and methods of cultivation on the production and viability of entomopathogenic fungi, Verticillium lecanii (Zimm.) Viegas

For mass production of V. lecanii, three types of cultivation methods including liquid, solid and diphasic production systems were investigated. In the liquid state of production, six media were tested in stationary culture conditions. Among the six media tested, Molasses Yeast Broth (MYB) supported maximum sporulation (8.33 x 10(8) spores mL(-1)) and biomass production (746 mg/100 mL). In the MYB, 4% molasses concentration was found to produce highest spore count (8.56 x 10(8) spores mL(-1)) and biomass (776 mg/100 mL) followed by 5 and 6% molasses. Among the six solid substrates tested, rice grains supported highest spore production (1.14 g/100 g). In diphasic state of production, combination of MYB and rice grains produced the greatest amount of spores, (1.70 g/100 g). Results of this study indicated that diphasic method using MYB as liquid medium and rice as solid substrate are the best method and media for mass production of V1-7 isolate of V. lecanii.     

Association of Lecithin Cholesterol Acyltransferase rs5923 Polymorphism in Iranian Individuals with Extremely Low High-Density Lipoprotein Cholesterol: Tehran Lipid and Glucose Study

Background:

The serum concentration of high-density lipoprotein cholesterol (HDL-C) is one of the important heritable risk factors for cardiovascular disease and is a target for therapeutic intervention. In this study, we aimed to evaluate the effects of lecithin cholesterol acyltransferase (LCAT) gene polymorphism rs5923 on LCAT enzyme activity and serum HDL-C concentration.

Methods:

The study population was selected from consecutive individuals with HDL-C ≤ 5^th percentile (n = 73) and extremely high HDL-C ≥ 95^th percentile (n = 57) who had participated in the Tehran Lipid and Glucose Study. The rs5923 polymorphism was genotyped using direct sequencing. LCAT activity was measured by fluorometric assay kit, and lipid concentrations were measured using the enzymatic colorimetric method.

Results:

The genotype frequencies were significantly different between the high HDL-C group (CC 94.7%, CT 5.3%) and the low HDL-C group (CC 83.6%, CT 16.4%) (P = 0.048). The T-allele frequencies in subjects with low and high HDL-C were 0.082 and 0.026, respectively (P = 0.16). The association of the single-nucleotide polymorphism rs5923 with low HDL-C was not statistically significant after adjustment for age, sex, and BMI (odd ratio = 2.65, 95% confidence interval = 0.32-21.5, P = 0.36, regression logistic analysis). Also, the effects of LCAT enzyme activity did not depend on the HDL-C level (P = 0.24).

Conclusion:

rs5923 polymorphism is not associated with low HDL-C levels in Iranian population. Key Words: Polymorphism, Single nucleotide, Lipoproteins     

Bioresource Efficacy of Phosphate Rock,Sulfur, and Thiobacillus Inoculum in Improving Soil Phosphorus Availability

In order to improve the effectiveness of phosphate rock as phosphorus fertilizer, elemental sulfur and Thiobacillus have been evaluated as amendments. First, Thiobacillus was isolated from different soil samples. Then, a greenhouse pot experiment was conducted using a completely randomized factorial design with three factors included: elemental sulfur at four levels of 0, 1000, 2000, and 5000 mg kg^?1; phosphate rock at three levels of 0, 1000 and 2000 mg kg^?1; four Thiobacillus inoculums (T₁, T₂, T₃, T₄) and without inoculation (T₀) in three replications. Results showed that all the four Thiobacillus inoculums increased significantly extractable soil-P. Combined application of phosphate rock and sulfur in equal proportion (1:1) along with inoculum Thiobacillus had a significant effect in improving phosphorus availability in soil. Combined application of sulfur (at rates of 1000 and 2000 mg kg^–1) and Thiobacillus significantly increased phosphorus uptake by plants as compared to the control.     

Effect of dietary phospholipid sources and levels on growth performance,enzymes activity,cholecystokinin and lipoprotein fractions of rainbow trout (Oncorhynchus mykiss) fry

This study was conducted to evaluate the effect of soybean lecithin (SBL) and chicken egg lecithin (EGL) on growth performance; pancreatic and intestine enzymes, cholecystokinin (CCK) and lipoprotein fractions of rainbow trout, Oncorhynchus mykiss (Walbaum) fry. Seven isonitrogenic and isolipidic diets were prepared by replacing 0 (as a control), 2%, 4% and 6% soybean and chicken egg lecithin instead of soybean oil in diets. Trout fry with initial average weight of 120 ± 4.08 mg, were randomly distributed in 21 90 L tanks. Triplicate groups (165 fish for each tank) of fry were fed each diet to visual satiation at six meals per day for 40 days. Dietary supplements of 2% and 4% chicken egg lecithin led to increase in fry final weight, weight gain and specific growth rate suggesting that moderate PL levels are needed during this stage of rainbow trout. Also 2% chicken egg lecithin caused significantly higher activity of amylase, lipase, phospholipase A₂, alkaline phosphatase (AP), aminopeptidase‐N (AN) and lower activity of leucine alanine peptidase (leu‐ala) in the digestive tract of fry than those in control group. Furthermore, fry fed 2% chicken egg lecithin had a higher ratio of AN/leu‐ala and AP/leu‐ala than that in the control group. Chicken egg lecithin increased significantly cholecystokinin (CCK) and chylomicron fractions of fry compared to control group. It is noteworthy that soybean lecithin had moderate effect on final growth, weight gain and specific growth rate of fry and induced significantly higher activity of amylase, lipase, phospholipase A₂ and secretion of CCK compared to control group. The results of this study indicate that dietary supplementation 2% chicken egg lecithin indirectly through some biochemical parameters improves growth performance of rainbow trout fry.     

Analysis of genetic and virulence variability of Stemphylium lycopersici associated with leaf spot of vegetable crops

Stemphylium lycopersici (Enjoji) W. Yamam was initially described from tomato and has been reported to infect different hosts worldwide. Sequence analyses of the internal transcribed spacer (ITS) regions 1 and 2, including 5.8S rDNA (ITS-5.8S rDNA) and glyceraldehyde-3-phosphate dehydrogenase (gpd) gene, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR), as well as virulence studies were conducted to analyze 46?S. lycopersici isolates. Stemphylium lycopersici isolates used in this study were obtained from diseased tomato (Solanum lycopersicum L.), eggplant (Solanum melongena L.), pepper (Capsicum annuum L.) and lettuce (Lactuca sativa L.) from major vegetable growing regions of Malaysia, including the three states of Pahang, Johor and Selangor between 2011 and 2012. Phylogenetic analysis of a combined dataset of the ITS-5.8S rDNA and gpd regions indicated that all isolates were clustered in the sub-cluster that comprised S. lycopersici, and were distinguished from other Stemphylium species. Cluster analyses using the UPGMA method for both RAPD and ISSR markers grouped S. lycopersici isolates into three main clusters with similarity index values of 67 and 68 %. The genetic diversity data confirmed that isolates of S. lycopersici are in concordance to host plants, and not geographical origin of the isolates. All S. lycopersici isolates were pathogenic on their original host plants and showed leaf spot symptoms; however, virulence variability was observed among the isolates. In cross-inoculation assays, the representative isolates were able to cause leaf spot symptoms on eggplant, pepper, lettuce and tomato, but not on cabbage.     

Tunable functional properties on polyester fabric using simultaneous green reduction of graphene oxide and silver nitrate

Here, a novel method is introduced to create tunable properties on the polyester fabric through diverse chemical modifications. The polyester fabric was primarily modified with NaOH or ethylenediamine to enhance the surface activity. This will produce diverse chemical groups on the polyester fabric surface including carboxylate, hydroxyl and amine groups. The fabric was treated with grahene oxide through exhaustion method. The silver nitrate was then added and simultaneously reduced with grapheme oxide using ascorbic acid and ammonia to produce reduced graphen oxide/silver nanocomposites (rGO/Ag) on the fabric surface. The synthesized nanocomposites were characterized by TEM and Raman spectra. The presence and uniform distribution of the nanocomposites on the fabric surface was also confirmed by SEM images and EDX patterns. The electrical resistivity was varied on the raw and modified polyester fabric due to the diverse formation of the graphene nanosheets network on the fabric surface. More Ag particles were formed on the surface of the alkali hydrolyzed polyester whereas more graphene nanosheets deposited on the aminolyzed polyester fabric. Also the hydrolyzed polyester fabric exhibited higher antibacterial properties with the lowest silver nitrate in the processing solution. The aminolyzed fabric showed a lower electrical resistance than the hydrolyzed and raw fabrics with the same amount of GO in the procedure bath. The aminolyzed polyester fabric indicated higher affinity towards GO produced higher antibacterial properties before reduction and without silver nitrate however lower electrical resistance obtained after reduction comparing with other samples.     

Mercury and methyl mercury concentration in sediment,benthic, Barbus Grypus and pelagic,Barbus esocinus fish species,from Musa estuary,Iran

The concentrations of mercury (Hg) and methyl mercury (MeHg) in sediment and tissues of Barbus grypus and Barbus esocinus fish species from Musa estuary, north part of the Persian Gulf, were measured. The order of Hg and MeHg concentrations in the sediment and tissues of the fish species was as follows: liver > gill > muscle > sediment. In the tissues of two fish species, the Hg and MeHg concentrations were highest in liver whereas lowest in the muscle. In the sediment, mean concentrations of Hg and MeHg were 0.83 and 0.437 µg g^?1. Mean concentrations of Hg and MeHg in B. grypus were 1.2–0.624 µg g^?1 and in all tissues of B. esocinus were 0.75 and 0.421 µg g^?1. Significant correlation between heavy metal concentration in sediment and fish may be related to high variability of heavy metal in the sediment.