Optimising cutting method and timing for the control of Abutilon theophrasti seed production

Abutilon theophrasti (velvetleaf) is a widespread and problematic annual weed. Greenhouse and field experiments were conducted to investigate the effects of different cutting methods on the viability of A. theophrasti seeds. Three cutting methods were assessed: (1) Entire plant cut and dried (EPD)—plants were cut at soil level and dried with capsules attached on the greenhouse bench or soil surface for 4 weeks; (2) capsules detached and dried (CD)—capsules were removed from plants and dried for 4 weeks; and (3) capsules detached and tested while fresh (CF)—a control treatment. Before drying, the developmental stage (stage one, dark green; stage two, light green; stage three, yellowish-green; or stage four, black with the slightly open capsule) and age (days after flowering, DAF) of each capsule was recorded. Seed viability was measured immediately in the CF treatment and after the 4-week drying period in the EPD and CD treatments. No seeds in the EPD and CD treatments were viable when harvested at the first developmental stage (1–8 DAF) in either experiment, but 100% of seeds in the CF treatment in the field were viable when harvested at 8 DAF. In both greenhouse and field experiments, seeds attained full viability at earlier harvest ages in CF than in EPD or CD treatments, suggesting that seeds might become viable relatively early in development but lose viability if allowed to dry. These findings could be applied to optimise late-season mechanical control of A. theophrasti.     

Nonparametric phenotypic stability analysis in advanced barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) genotypes

The development of genotypes with adaptation to a wide range of environments is one of the most important goals of plant breeding programs. In order to compare nonparametric stability measures and to identify promising high-yield and stable barley (Hordeum vulgare L.), 20 barley genotypes selected from the Iran/ICARDA joint project and grown in nine environments during 2009-11 in Iran. Four nonparametric statistical tests of significance for genotype × environment (GE) interaction and 10 nonparametric measures of stability were used to identify stable genotypes in nine environments. Results of nonparametric tests of G×E interaction (Kubinger, Hildebrand, and Kroon/ Laan) and a combined ANOVA across environments, indicated the presence of both crossover and non-crossover interactions. Also, only TOP and rank-sum values were positively associated with high yield. Thus, in the simultaneous selection for high yield and stability, only the rank-sum and TOP methods were useful in terms of the principal component analysis results, and correlation analysis of nonparametric stability statistics and yield. According to these stability parameters (TOP and rank-sum), three genotypes (G13, G12, and G17) were the most stable for grain yield. The results also revealed that based on nonparametric test results, stability could be classified into three groups, according to agronomic and biological concepts of stability.     

Shortening the generation cycle in faba bean (Vicia faba) by application of cytokinin and cold stress to assist speed breeding

The aim of this study was to reduce the length of the breeding cycle for faba bean by accelerating seed setting. We examined the mode and time of exogenous 6-benzylaminopurine (BAP) (cytokinin) application, and cold treatments and their combinations in two faba bean genotypes. Acropetal node number of pod and seed set and pollen viability were recorded during the experiments. Application of BAP improved pollen germination. The application of 10^–5 M BAP 4 days after flowering increased seed set at the lower nodes. Cold treatment (8/4°C day/night for 2 days) after the onset of flowering induced the formation of more pods and faster pod set compared to the non-cold treatment. The time to first seed was significantly reduced, and pollen viability was increased in plants exposed to cold treatment. Increased pollen viability also showed a significant positive correlation with seed setting. The combinations of 10^–5 BAP and cold treatment together had similar and independent effects. These results will accelerate plant breeding in faba bean by providing additional tools for reducing generation time.     

Effects of emulsified versus saline administration of GnRHa on induction of ovulation in rainbow trout, Oncorhynchus mykiss

The effects of saline-dissolved or Freund's incomplete adjuvant (FIA)-emulsified GnRHa treatment on the induction of ovulation in rainbow trout, Oncorhynchus mykiss, were examined. The FIA-emulsified GnRHa was first diluted in 0.25 ml physiological saline and then mixed with an equal volume of FIA. Fish were selected in the beginning of the spawning season and were allocated into four groups and were treated intraperitoneally with (a) 0.5 ml of emulsified GnRHa (GnRHa–FIA), (b) 0.5 ml saline-dissolved GnRHa in a single injection (GnRHa-1), (c) 0.5 ml saline-dissolved GnRHa in two injections spaced 1 d apart (GnRHa-2) and (d) 0.5 ml of saline (Control). The GnRHa dose in all hormone treatments was 25 µg kg^− 1. All fish in the FIA–GnRHa and GnRHa-2 groups ovulated within 10 and 11 d after treatment, respectively. In contrast, only 75% in the Control fish and 60% of the fish in the GnRHa-1 group ovulated within 36 d after treatment. None of the treatments caused any pre- or post-spawning mortality in the broodstock. Fertilization, eyeing and hatching percentages of the produced progeny were normal in all the treatment groups and did not differ significantly among them. In conclusion, FIA-emulsified GnRHa can be effective in advancing the onset of and synchronizing the ovulation of rainbow trout within a two-week period, thus shortening the egg collection period, without affecting broodstock survival and egg quality.     

Effects of the prebiotic mannan‐oligosaccharide on feed deprived zebrafish: Growth and reproduction

This study was designed to investigate whether the prebiotic mannan‐oligosaccharides (MOS) can reduce the deleterious impacts of feed deprivation on growth and reproduction in zebrafish. In the growth performance experiment, juvenile fish were distributed in the following four treatments: normal‐control (NC), starved‐control (SC), normal‐prebiotic (NP) and starved‐prebiotic (SP). After 8 weeks, NP and SC fish showed the highest and lowest growth patterns, respectively among treatments. Standard length, specific growth rate and feed conversion ratio did not differ significantly between NC and SP treatments. Autochthonous lactic acid bacteria (LAB) were significantly higher in NP and SP than the NC and SP treatments. Feed restriction resulted in significantly lower concentrations of thyroxine. In the reproductive performance experiment, additional juveniles were fed as in the growth experiment. Crossing in the final week of the experiment resulted in successful spawning in only NP fish, which showed mature sperm and oocytes in histological examinations. The number of spermatozoa was significantly lower in the fish that experienced feed restriction; however, the oocytes of SP females were at the same maturation level to that observed in NC females. Sex‐steroids changed after both starvation and MOS supplementation where NC and SP showed no differences in the levels of testosterone and female's 17β‐estradiol. These results indicate that MOS supplemented diet reduced some side effects of feed deprivation (final weight and length, SGR, FCR and levels of LAB and T3), and suggest that supplementation of the diet with MOS may ameliorate some of the negative effects of feed deprivation in zebrafish.     

Rapid one-step separation and purification of recombinant phenylalanine dehydrogenase in aqueous two-phase systems

Background: Phenylalanine dehydrogenase (PheDH; EC 1.4.1.20) is a NAD+-dependent enzyme that performs the reversible oxidative deamination of L-phenylalanine to phenylpyruvate. It plays an important role in detection and screening of phenylketonuria (PKU) diseases and production of chiral intermediates as well. The main goal of this study was to find a simple and rapid alternative method for purifying PheDH. Methods: The purification of recombinant Bacillus sphaericus PheDH was investigated in polyethylene glycol (PEG) and ammonium sulfate aqueous two-phase systems (ATPS). The influences of system parameters including PEG molecular weight and concentration, pH and (NH4)2SO4 concentration on enzyme partitioning were also studied. The purity of enzyme was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Results: A single extraction process was developed for separation and purification of recombinant PheDH from E. coli BL21 (DE3). The optimized conditions for partitioning and purification of PheDH were 9% (w/w) PEG-6,000 and 16% (w/w) (NH4)2SO4 at pH 8.0. The partition coefficient, recovery, yield, purification factor and specific activity values were achieved 58.7, 135%, 94.42%, 491.93 and 9828.88 U/mg, respectively. Also, the Km values for L-phenylalanine and NAD+ in oxidative deamination were 0.21 and 0.13 mM, respectively. Conclusion: The data presented in this paper demonstrated the potential of ATPS as a versatile and scaleable process for downstream processing of recombinant PheDH.     

Western blot analysis of Leishmania infantum antigens using sera from pentamidine-treated dogs

Pentamidine treatment has been used successfully to establish immune cellular responses in recovered dogs. In this paper, we examined the appearance and disappearance of antibodies over time against crude Leishmania antigens and purified gp63 or gp70 proteins in sera from cured dogs using a Western blotting technique.Following the treatment, a pattern of antibody specificities to parasite antigens was observed in the sera of cured dogs. Antibodies to gp63 and gp70 were maintained after cure. In addition, the reaction with a 26 kD band observed during the clinical phase was no longer recognized by sera taken from recovery dogs. Interestingly, two proteins, 85 and 110 kD, not observed during the patent phase were detected by sera taken from treated dogs. Such patterns of antibody specificities to various parasite antigens might represent a useful parameter to determine the actual phase of the disease process.     

Streptomyces sp., a cause of fistulous withers in donkeys

Four of 10 donkeys, which showed lesions simulating fistulous withers, were examined clinically with the aim to cultivate and identify the causal agent. Aspiated purulent materials were subjected to bacteriological examination. The causal organisms were recovered in Tryptic Soya agar medium when incubated aerobically at 37 degrees C for up to 5 days. These organisms were found to be actinomycetes-like, Gam positive with stable branching filaments and to form heavy aerial hyphae on colony surface. The isolated organisms ere tentatively identified as Streptomyces sp. on the basis of morphological and cultural characteristics. The initial sequences analysis of the 16S rDNA gene conformed that one of the isolates (SD551) falls within the phylogenetic clade, which encompasses the genus Streptomyces. Studies are underway to further describe the disease and its causal agent. The report represents a good evidence to incriminate Streptomyces in the aetiology of the fistulous withers.     

Construction of a microarray specific to the chicken immune system: profiling gene expression in B cells after lipopolysaccharide stimulation

The objective of this study was to profile gene expression in cells of the chicken immune system. A low-density immune-specific microarray was constructed that contained genes with known functions in the chicken immune system, in addition to chicken-expressed sequence tags (ESTs) homologous with mammalian immune system genes, which were systematically characterized by bioinformatic analyses. Genes and ESTs that met the annotation criteria were amplified and placed on a microarray. The microarray contained 84 immune system gene elements. As a means of calibration, the microarray was then used to examine gene expression in chicken B cells after lipopolysaccharide stimulation. Differential gene expression was observed at 6, 12, and 24 h but not at 48 h after stimulation. The results were validated by semiquantitative polymerase chain reaction. The microarray showed a high degree of reproducibility, as demonstrated by intra- and interassay correlation coefficients of 0.97 and 0.95, respectively. Thus, the low-density microarray developed in this study may be used as a tool for monitoring gene expression in the chicken immune system.     

The incidence of canine haematozoa in peninsular Malaysia

In 3 urban areas in Selangor, Peninsular Malaysia between 1973 and 1981, blood from 4080 dogs was examined for haematozoa. The following frequencies were found: Babesia gibsoni 17.7%; microfilariae of Dirofilaria immitis 9.6%; Hepatozoon canis 1.2%; B. canis 1.1%; Ehrlichia canis 0.2%; Trypanosoma evansi 0.1%. A detailed examination of B. gibsoni infections and microfilariasis due to D. immitis with regards to monthly distribution, breed frequency, sex and age, revealed that pedigree and non-pedigree dogs were equally susceptible to Babesia and microfilariae infections.