Application of the consolidated species concept to identify the causal agent of strawberry anthracnose in Iran and initial molecular dating of the <Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> species complex

Anthracnose caused by Colletotrichum spp. is one of main strawberry diseases worldwide. The disease recently arrived in Iran and become epidemic in the Kurdistan province. A combination of morphological, molecular and pathogen-host interaction approaches (referred to the consolidated species concept) were applied to isolates from symptomatic strawberry tissues collected in Iran. Multi-gene phylogenetic analysis based on ITS-rDNA, beta-tubulin (TUB2) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) identified the species as the S5 clade of Colletotrichum nymphaeae. No significant intraspecies variation was observed in terms of morphology and pathogenicity. No significant genetic variation was observed among the isolates using inter simple sequence repeat analysis (ISSR) and primer pair combination of ERIC1/BOX and ERIC2/BOX fingerprinting markers. Based on the results of this study, it can be postulated that the C. nymphaeae population in Iran established itself from a single origin due to a founder effect. More generally, molecular dating based on relaxed clocks indicates co-radiation of C. nymphaeae S5 and strawberry plants and suggests high reciprocal specificity between the host and pathogen.     

A Molecular Survey of Campylobacter jejuni and Campylobacter Coli Virulence and Diversity

Background: The aim of this study was to determine the prevalence of virulence-associated genes and enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) analysis of Campylobacter spp. isolated from children with diarrhea in Iran. Methods: A total of 200 stool specimens were obtained from children under 5 years during July 2012 to July 2013. Detection of C. jejuni and C. coli was performed by standard biochemical and molecular methods. The presence of virulence-associated genes and genetic diversity of isolates was examined using PCR and ERIC-PCR analyses. Results: A total of 12 (6%) Campylobacter spp. were isolated from patients including 10 (4.5%) C. jejuni and 2 (1.5%) C.coli. The flaA, cadF and ciaB genes were present in 100% of isolates, while no plasmid of virB11 gene was present in their genome. The prevalence of invasion-associated marker was 100% among C. coli and was not detected in C. jejuni isolates. The distribution of both pldA and the genes associated with cytolethal distending toxin (CDT) was 58.3% in C. jejuni isolates. Seven distinct ERIC-PCR profiles were distinguished in three clusters using ERIC-PCR analysis. Genotyping analysis showed a relative correlation with geographic location of patients and virulence gene content of isolates. Conclusion: To our knowledge, this is the first molecular survey of Campylobacter spp. in Iran concerning genotyping and virulence gene content of both C. jejuni and C. coli. ERIC-PCR revealed appropriate discriminatory power for clustering C. jejuni isolates with identical virulence gene content. However, more studies are needed to clearly understand the pathogenesis properties of specific genotypes. Key Words: Campylobacter jejuni, Campylobacter coli, Ddiarrhea, Virulence factors     

Effect of microwave–chemical pre-treatment on compression characteristics of biomass grinds

The effect of microwave and microwave–chemical pre-treatments on densification characteristics and physical quality of pellets made from wheat and barley straw grinds were investigated. The ground wheat and barley straw samples were immersed in water, sodium hydroxide or calcium hydroxide solution at different concentrations (1 and 2% w/v) and then exposed to microwave radiation at three power levels (295, 603 and 713 W). Chemical composition and bulk and particle densities of samples were determined after pre-treatments. Pre-treated grinds were compressed in a plunger–die assembly with a force of 4000 N and compression and relaxation test data were recorded. The specific energy required for compression and ejection of pellets produced from untreated and pre-treated wheat and barley straw grinds was calculated. The tensile strength of the pellets was also evaluated to investigate the hardness of the pellets. Chemical composition analysis showed that microwave and chemical pre-treatment was significantly able to disintegrate the lignocellulosic structure of wheat and barley straw grinds. Data analysis also indicated that the pellets made from microwave–chemical pre-treated biomass grinds had a significantly higher density and tensile strength than the untreated or samples pre-treated by microwave and distilled water.     

Zinc sorption–desorption by sand,silt and clay fractions in calcareous soils of Iran

Zinc sorption–desorption by sand, silt and clay fractions of six representative calcareous soils of Iran were measured. Sand, silt and clay particles were fractionated after dispersion of soils with an ultrasonic probe. Zinc sorption analysis was performed by adding eight rates of Zn from 6 to 120 μmol g^?1. For the desorption experiment, samples retained after the measurement of Zn sorption were resuspended sequentially in 0.01 M NaNO₃ solution and shaken for 24 h. Results indicated that Zn sorption by soil fractions increased in the order clay > silt > sand, and correlated negatively with CaCO₃ content and positively with cation exchange capacity (CEC) and smectite content. Results indicated that for all fractions, the Langmuir equation described the sorption rates fairly well. In contrast to sorption, Zn desorption from soil fractions increased in the order sand > silt > clay, and correlated positively with CaCO₃ content, CEC and smectite content. Results showed that parabolic diffusion and two constant equations adequately described the reaction rates of Zn desorption. In general, for all soils studied, the coarser the particle size, the less Zn sorption and more Zn desorption, and this reflects much higher risk of Zn leaching into groundwater or plant uptake in contaminated soils.     

Effect of hydrothermaled bran on physicochemical, rheological and microstructural characteristics of Sangak bread

Increased consumption of whole grain products has been associated with decreased risk of health problems such as cardiovascular disease, diabetes and obesity. Phytate however, has always been a matter of concern, since it chelates minerals such as iron. Sangak is very popular Iranian flat bread invented five hundred years ago, made from 95% extraction flour. The bread is fermented but usually fermentation is not able to remove all phytate from the dough. In this study flours were first debranned; resulting bran was subjected to a hydrothermal process. The hydrothermaled (HT) brans were then incorporated in the Sangak flour. In the next step, physicochemical, rheological and microstructural characteristics of Sangak flour and dough prepared from two Iranian wheat varieties, Tajan and Back Cross of Roshan were investigated. Results indicated a reduction in phytate up to 55% in the samples. The resulting dough containing HT bran showed a higher development time and valorimetric value and was more stable than doughs made with normal bran. Dough made with HT bran showed a kind of protein matrix in which proteins and starch granules are oriented in a more non-ordered structure.     

Silver nitrate and adenine sulphate induced high regeneration frequency in the recalcitrant plant Cosmos bipinnatus using cotyledon explants

Plant regeneration ability was studied in the medicinal-ornamental plant, Cosmos bipinnatus ‘Sonata white’, which is a dicotyledonous recalcitrant plant to shoot induction. Cotyledons were used as sources of explants to investigate plant regeneration. High frequency of direct shoot induction was obtained when BA (5 mg/l) and AgNO₃ (5 mg/l) were used in combination with 20 mg/l adenine sulphate (73.8%) in Murashige and Skoog (MS) medium. The highest shoot number per explant (5.7) was induced on MS medium supplemented with 5 mg/l BA, 5 mg/l AgNO₃, and 40 mg/l adenine. Eight week-old shoots were transferred to root induction media containing MS and half-strength MS medium with different concentration of IBA. The highest rate of root induction (70.8%) was obtained on half-strength MS medium with 1.5 mg/l IBA within four weeks. The plantlets were transferred to pot and kept in the greenhouse condition. Seventy percent of the plantlets successfully acclimatised.

Abbreviations: BA, 6-benzylaminopurine; IBA, Indole-3-butyric acid; MS, Murashige and Skoog; PGRs, plant growth regulators.     

Prevotella copri,a potential indicator for high feed efficiency in western steers

There has been a great interest to identify a microbial marker that can be used to predict feed efficiency of beef cattle. Such a marker, specifically one that would allow an early identification of animals with high feed efficiency for future breeding efforts, would facilitate increasing the profitability of cattle operations and simultaneously render them more sustainable by reducing their methane footprint. The work presented here suggests that Prevotella copri might be an ideal microbial marker for identifying beef cattle with high feed efficiency early in their life span and in the production cycle. Developing more refined quantification techniques that allow correlation of P. copri to feed efficiency of beef cattle that can be applied by lay people in the field holds great promise to improve the economy of cattle operations while simultaneously reducing their environmental impact by mitigating methane production from enteric fermentation.     

Effects of blend of canola oil and palm oil on nutrient intake and digestibility,growth performance,rumen fermentation and fatty acids in goats

The study examined the effects of blend of 80% canola oil and 20% palm oil (BCPO) on nutrient intake and digestibility, growth performance, rumen fermentation and fatty acids (FA) in goats. Twenty‐four Boer bucks were randomly assigned to diets containing 0, 4 and 8% BCPO on a dry matter basis, fed for 100 days and slaughtered. Diet did not affect feed efficiency, growth performance, intake and digestibility of all nutrients except ether extract. Intakes and digestibilities of ether extract, unsaturated fatty acids (FA) and total FA were higher (P < 0.05) while digestibility of C18:0 was lower (P < 0.05) in oil‐fed goats than the control goats. Total volatile FA, acetate, butyrate, acetate/propionate ratio and methane decreased (P < 0.05) with increasing BCPO but propionate, NH₃‐N and rumen pH did not differ between diets. Ruminal concentration of C18:0, n‐3 FA and total FA increased (P < 0.05) while C12:0, C14:0, C15:0 and n‐6 FA decreased with increasing BCPO. Analysis of the FA composition of Triceps brachii muscle showed that concentrations of C16:0, C14:0 and C18:2n‐6 were lower (P < 0.05) while C18:1n‐9, C18:3n‐3 and C20:5n‐3 were higher in oil‐fed goats compared with control goats. Dietary BCPO altered muscle lipids without having detrimental effects on nutrient intake and digestibility and growth performance in goats.     

Cloning and expression of coxsakievirus B3 viral protein-1 in E. coli

BACKGROUND: Viral protein-1 (VP1) is a major capsid protein of Coxsakievirus B3 (CVB3) that plays an important role in directing viruses towards permissive cells and acts as a main antigenic site of the virus in eliciting of host immune response, hence it seems VP1 can be considered as a vaccine candidate against CVB3 infection. In this study, cDNA of VP1 was prepared, cloned into pET expression vector and the recombinant protein (VP1) was over expressed in E. coli. METHODS: The viruses were grown in suspension cultures of Vero cells with an input virus multiplicity of 10-50 plaque-forming units/cell. After observing complete cytopathic effect, the total RNA (cells and virus) was prepared for RT-PCR and by using specific primers, VP1 cDNA was amplified and ligated into pET vectors (32 a and 28 a). The recombinant vector was transferred into competent E. coli (BL-21) and after selection of proper colony, which carried correct cDNA within the vector; cells were cultured and induced with isopropyl B-D-thiogalactopyranoside, in order to express protein (VP1). The cultures were tested for presence of VP1 by SDS-PAGE and Western-Blotting analysis. RESULTS: Molecular techniques such as PCR which showed exact defined size of the VP1 (819 bp), restriction digestion and finally immunoblot analysis of over expressed protein; all confirmed the correct cloning and expression of VP1 in this research. CONCLUSION: In this research, full length of VP1 as major capsid protein of CVB3 was over expressed in E. coli which, can be used for further studies, including neutralizing antibody production against CVB3.