Identification of Arcanobacterium (Trueperella) abortisuis, a novel species of veterinary importance, by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)

In the present study A. (T.) abortisuis isolated from pigs and bovines could be reliably identified by determination of phenotypic properties, genotypically by polymerase chain reaction with the help of A. (T.) abortisuis 16s-23S rDNA intergenic spacer region specific oligonucleotide primer and by Matrix-Assisted Laser Desorption Ionization-Time Of Flight mass spectrometry (MALDI-TOF MS). The latter appeared to be a promising tool for fast and cost effective identification of this species and might help to elucidate the role A. (T.) abortisuis plays in infections of pigs, bovines, possibly other animals or humans.     

Impact of 12-year field treatments with organic and inorganic fertilizers on crop productivity and mycorrhizal community structure

The effect of manure and mineral fertilization on the arbuscular mycorrhizal (AM) fungal community structure of sunflower (Helianthus annuus L.) plants was studied. Soils were collected from a field experiment treated for 12 years with equivalent nitrogen (N) doses of inorganic N, dairy manure slurry, or without N fertilization. Fresh roots of tall fescue (Festuca arundinacea Schreb.) grass collected from the field plots without N fertilization and unfumigated field soils were used as native microbial inoculum sources. Sunflower plants were sown in pots containing these soils, and three different means of manipulating the microbial community were set: unfumigated soil with fresh grass roots, fumigated soil with fresh grass roots, or fumigated soil with sterilized grass roots. Assessing the implications with respect to plant productivity and mycorrhizal community structure was investigated. Twelve AM fungal OTUs were identified from root or soil samples as different taxa of Acaulospora, Claroideoglomus, Funneliformis, Rhizophagus, and uncultured Glomus, using PCR-DGGE and sequencing of an 18S rRNA gene fragment. Sunflower plants grown in manure-fertilized soils had a distinct AMF community structure from plants either fertilized with mineral N or unfertilized, with an abundance of Rhizophagus intraradices-like (B2). The results also showed that AM inoculation increased P and N contents in inorganic N-fertilized or unfertilized plants, but not in manure-fertilized plants.     

The Effect of Vitrification on Mouse Oocyte Apoptosis by Cryotop Method

Background : Oocyte cryopreservation is one of the most important topics in the field of assisted reproductive technology to preserve women fertility, but relationship between cryopreservation and apoptosis is still a matter of debate. The present study was aimed to investigate the effects of vitrification on apoptosis in mouse oocytes by Cryotop method. Method: A total of 200 germinal vesicle (GV) and 200 metaphase II (MII) oocytes were obtained from ovaries and fallopian tubes of NMRI mice, respectively and divided into control and experimental groups. Oocytes in experimental group were vitrified by Cryotop using vitrification medium and were kept in liquid nitrogen for one month. The survival rate of oocytes was evaluated after 2 hour incubation time. Then, the oocyte apoptosis was evaluated by TUNEL technique and compared with those in control group. The data was compared statistically using SPSS software and chi-square test. Results: The survival rates of vitrified GV (93%) and MII oocytes (88%) showed a significant decrease compared with the control group (P<0.05), but there was no significant difference in survival rate of both vitrified oocyte groups. The incidence of apoptosis in vitrified and control GV oocytes showed no significant difference (13% vs. 7%), but the rate of apoptosis in vitrified MII oocytes increased significantly not only in comparison with MII control group (25% vs. 5%) but also with vitrified GV oocytes (P<0.05). Conclusion: The results indicate that vitrification increases apoptosis in mouse MII oocytes and apoptosis may play a role in MII oocyte injury after vitrification. Key Words: Vitrification, Apoptosis, Oocytes     

Pathogenicity determinants in smut fungi revealed by genome comparison

Biotrophic pathogens, such as the related maize pathogenic fungi Ustilago maydis and Sporisorium reilianum, establish an intimate relationship with their hosts by secreting protein effectors. Because secreted effectors interacting with plant proteins should rapidly evolve, we identified variable genomic regions by sequencing the genome of S. reilianum and comparing it with the U. maydis genome. We detected 43 regions of low sequence conservation in otherwise well-conserved syntenic genomes. These regions primarily encode secreted effectors and include previously identified virulence clusters. By deletion analysis in U. maydis, we demonstrate a role in virulence for four previously unknown diversity regions. This highlights the power of comparative genomics of closely related species for identification of virulence determinants.     

Thiamine (vitamin B1) deficiency in germinating seeds of Phaseolus vulgaris L. exposed to soaking injury

Exogenous application of thiamine (vitamin B1) during imbibition improved germination and seedling development of bean (Phaseolus vulgaris L. cv. Lasso) seeds, which were exposed to soaking injury by submergence. Leaching of the vitamin into the incubation medium was not increased in submerged seeds. However, translocation of the thiamine reserves from the cotyledons to the seedling axis was reduced in response to soaking injury. After exogenous application of free thiamine to the seeds, thiaminedi-phosphate (TDP), which is the physiologically active coenzyme form of thiamine, accumulated in the seedling axis of submerged seeds, suggesting an increased demand for thiamine-dependent metabolic reactions in these tissues. Limited oxygen supply of the seeds during the soaking period induces a shift from respiration to fermentation of carbohydrates. Neither thiamine-dependent ethanolic fermentation, nor ATP production or adenylate energy charge (AEC) of the seedling axis were affected by exogenous thiamine application, suggesting that there is no limitation of thiamine-dependent reactions in the energy metabolism of the seedlings. Thus the physiological mechanisms improving germination and seedling development of submerged seeds in response to thiamine seed treatment are still not clear.     

Non‐specific immune responses and intestinal immunity of common carp (Cyprinus carpio) fed Jujube (Ziziphus jujube) fruit extract

This study investigated the effects of Jujube (Ziziphus jujube) fruit extract (JFE) as an excellent source of nutrients and phytochemicals on the innate immune responses and expression of genes involved in intestinal immunity of common carp (Cyprinus carpio) fingerlings. Three hundred healthy carps (10.78 ± 0.05 g) fingerlings were randomly allocated to 12 fibre glass tanks (300L; 15 fish per tank) and fed for 8 weeks with experimental diets which contained different levels of JFE (0%, 0.25%, 0.5%, or 1%). Thereafter, serum non‐specific immune parameters [total immunoglobulin (Ig) as well as alternative complement (ACH50) and lysozyme activity (SL)] and expression of genes involved in intestinal immunity (lyz, tnf‐alpha, il1b, il8, il10, and tgfb) were measured. The results revealed a significant elevation of Ig level in the JFE‐fed fish (p < 0.05) over the control fish. However, there was no significant difference among treated groups in case of serum total Ig. The highest levels of SL and ACH50 were observed in fish fed 1% of JFE. Nonetheless, no significant difference was noticed regarding SL and ACH50 of control and other supplemented groups (0.25% and 0.5% JFE). Also, expression of lyz, tnf‐alpha, and il1b genes increased significantly (p < 0.05) in JFE‐fed fish, in a dose‐dependent manner. On the other hand, significant (p < 0.05) down‐regulation of il110 and tgfb were noticed in treated groups. The present findings suggested that extraction of Jujube, Z. jujube fruit, possesses beneficial effects on immune responses of common carp.     

Effects of carbohydrate source and polyethylene glycol on maturation and germination of somatic embryos in walnut (<Emphasis Type="Italic">Juglans regia</Emphasis> L.)

Low efficiency of somatic embryo maturation, germination, and conversion to plantlets is a major problem in many species including J. regia L. Germination efficiency of somatic embryos is very low in walnut. In this study, effects of two carbohydrate sources, sucrose and maltose (each at 3% and 6%), and two kinds of PEG (4000 and 6000) (each at four levels of 1.5%, 3%, 5% and 7.5%) on maturation and germination of walnut embryos were tested. The number of somatic embryos increased conspicuously on medium containing PEG. Furthermore, higher levels of PEG-4000 (7.5%) could remarkably enhance the morphogenesis of somatic embryos and the number of embryos produced. PEG-4000 stimulated somatic embryo maturation of walnut. This stimulatory effect was dependent on the carbohydrate source used. Sucrose in combination with PEG-4000 produced 50% of cotyledonary and normal somatic embryos. Different concentrations of PEG were effective on the number of embryos with a shoot meristem. PEG-4000 7.5% and sucrose 3.0% produced the highest rate (50.0%) of normal shooting embryos. However, PEG (4000, 6000) and maltose caused an unfavorable effect and increased the frequency of abnormal shaped somatic embryos.     

Characterization of a Probiotic Lactobacillus fermentum Isolated from Snakehead,Channa striatus,Stomach

This study sought to isolate and characterize lactic acid bacteria (LAB) from stomach of adult snakehead fish, Channa striatus, to be used as probiotics for freshwater fish. A total of 13 strains were isolated from the stomach of 10 fish, and 4 of these belonged to LAB. Strain LAB‐3 showing highest in vitro growth inhibition of Aeromonas hydrophila in a disk diffusion test was identified as Lactobacillus fermentum by conventional and molecular techniques and evaluated in vitro through various tests. The bacterium could grow at pH 3–8; but the optimum growth was observed at pH 6. Moreover, LAB‐3 grew at 0.15 and 0.3% bile salt concentrations, from 15 to 45 C, and at 4% NaCl. L. fermentum showed in vitro inhibitory activity against three fish pathogens, A. hydrophila, Pseudomonas aeruginosa, and Shewanella putrefaciens, tested by disk diffusion and well diffusion methods. Antibiotic sensitivity tests indicated that L. fermentum was resistant to streptomycin, gentamycin, and kanamycin, intermediate to tetracycline, but sensitive to chloramphenicol, amoxicillin, and ampicillin. Challenge test by using A. hydrophila showed that survival of snakehead was significantly (P < 0.05) improved when 2 × 10⁶ LAB‐3/g was supplemented to the diet. Therefore, this study suggests that L. fermentum might be a promising probiotic in snakehead aquaculture .     

Identification of superior parents and hybrids for improving canola production under optimum and late sowing conditions

Major advancement in canola breeding depends on heterotic hybrids that require high general combining ability (GCA) and specific combining ability (SCA) inbred lines. In order to estimate heritability, gene action type, GCA, SCA and heterosis and to identify superior hybrids with wider adaptation to cold, one hundred canola hybrids were produced by crossing 10 lines and 10 testers in a Line?×?Tester mating design. The F₁ and F₂ generations were sown in α-lattice design in 2012 and 2013 growing seasons under optimum (early October) and late sowing (early November) conditions to be evaluated for days to flowering, days to physiological maturity, number of pods per plant, number of seeds per pod, thousand seed weight, seed yield and leaf electrical conductivity. The combined analysis indicated sufficient genetic diversity in the population and significant difference between two sowing date. The Line?×?Tester analysis presented significant GCA and SCA effects for all studied traits across optimum and late sowing conditions. The main gene action type was found to be non-additive, especially incomplete dominance and over-dominance in both conditions. Narrow-sense heritability ranged from low to moderate whereas broad-sense heritability was recorded more than 60% for all of the studied traits in both generations and conditions. The average heterosis in F₂ population for all studied traits was lower than that in F₁ representing this fact that heterosis is generally related to the heterozygosity at the population level and poorly correlated with heterozygosity at the individual level.     

Feed preference and growth response of juvenile Litopenaeus vannamei to supplementation of marine chemoattractants in a fishmeal‐challenged diet

This study compared the feed preference and growth response of Litopenaeus vannamei to chemoattractants. A diet with 3% fishmeal was supplemented with either 3% salmon meal (POS), 3% soy protein concentrate (NEG), 3% krill meal (KRM), 3% squid meal (SQM), 3% shrimp head meal (SHM), 3% shrimp meal (SM), 3% squid liver meal (SLM), or 5% liquid sardine hydrolysate (SAH). Shrimp with a body weight (BW) of 0.99 ± 0.08 g were stocked at 100 animals/m² in 56 tanks of 1 m³ and fed 10 times daily for 74 days. Feed preference was evaluated by feeding shrimp of 10.87 ± 1.82 g in excess twice a day for 10 days in two separate feeding trays allocated in 50 tanks of 0.5 m³. Survival reached 93.3 ± 5.80% and was unaffected by supplementation. Final BW was the highest for shrimp fed the KRM‐supplemented diet (11.97 ± 0.93 g), followed by POS (11.11 ± 0.77 g) and SQM (11.01 ± 1.17 g). Diets SHM, SM, SLM, and NEG showed a lower shrimp BW than POS, but were not statistically different among them. Shrimp fed the SAH diet achieved the lowest BW (10.06 ± 1.02 g). The highest gained yield was obtained with diets KRM and POS. No statistical difference was observed in shrimp yield among other diets. The lowest feed conversion ratio (FCR) was achieved with shrimp fed KRM (1.31 ± 0.05) when compared to diets SHM (1.47 ± 0.05), SAH (1.47 ± 0.07), and SLM (1.45 ± 0.17). Two‐by‐two comparisons indicated that shrimp preferred SHM and KRM, except when these were compared to SQM and SLM. No difference in feed preference was found between diets with SQM and SLM. SAH was the least preferred raw material in all comparisons. Results indicated that KRM acts as a powerful feeding effector and growth enhancer in fishmeal‐challenged diets for whiteleg shrimp. A dietary supplementation with 3% KRM is more effective than the same dose of any other chemoattractant evaluated.