Molecular and serotyping characterization of shiga toxogenic Escherichia coli associated with food collected from Saudi Arabia

Shiga toxin-producing Escherichia coli (STEC) strains are considered as one of the major food-borne disease agents in humans worldwide. STEC strains, also called verotoxin-producing E. coli strains. The objectives of the present study were serotyping and molecular characterization of shiga toxigenic E. coli associated with raw meat and milk samples collected from Riyadh, Saudi Arabia. A total of 540 milk samples were collected from 5 dairy farms and 150 raw meat samples were collected from different abattoirs located in Riyadh, Saudi Arabia. E. coli were recovered from 86 milk samples (15.93%), serotyping of E. coli isolates revealed, 26 (4.81%) strains O157: H7, 23 (4.26%) strains O111, 20 (3.70%) strains O113: H21, 10 (1.85%) strains O22: H8 and 7 (1.3%) strains O172: H21. Meanwhile, 17 (11.33%) strains of E. coli were recovered from raw meat samples, serotyping of E. coli isolates revealed, 6 (4%) strains O157: H7, 5 (3.33%) strains O111 and 4 (2.67%) strains O174: H2 and only two (1.33%) strains were identified as O22: H8. Shiga toxin2 was detected in 58 (67.44%) serotypes of E. coli recovered from milk samples and 16 (94.12%) serotypes of E. coli recovered from meat samples, while intimin gene was detected in 38 (44.186%) serotypes of E. coli recovered from milk samples and in 10 (58.82%) serotypes of E. coli recovered from meat samples. The results of this study revealed the efficiency of combination between serotyping and molecular typing of E. coli isolates recovered from food of animal origin for rapid detection and characterization of STEC.     

Detection and differentiation of the coconut lethal yellowing phytoplasma in coconut‐growing villages of Grand‐Lahou,Côte d'Ivoire

Surveys for the Côte d'Ivoire lethal yellowing (CILY) phytoplasma were conducted in eight severely CILY‐affected villages of Grand‐Lahou in 2015. Leaves, inflorescences and trunk borings were collected from coconut palms showing CILY symptoms and from symptomless trees. Total DNA was extracted from these samples and tested by nested polymerase chain reaction/RFLP and sequence analysis of the 16S rRNA, ribosomal protein (rp) and the translocation protein (secA) genes. The CILY phytoplasma was detected in 82.9% of the symptom‐bearing palms collected from all the surveyed villages and from all the plant parts. Trunk borings were recommended as the most suitable plant tissue type for sampling. Results indicate that the CILY phytoplasma may have a westward spread to other coconut‐growing areas of Grand‐Lahou. CILY phytoplasma strains infecting coconut palms in the western region of Grand‐Lahou exhibited unique single nucleotide polymorphisms on the rp sequence compared to the strains from the eastern region. Moreover, single nucleotide polymorphisms on the SecA sequence distinguished the CILY phytoplasma from the Cape St. Paul Wilt Disease phytoplasma in Ghana, and the Lethal Yellowing phytoplasma in Mozambique.     

Controlling the sugar beet fly Pegomyia mixta Vill. with entomopathogenic nematodes

Sugar beet, Beta vulgaris L. is a strategic crop of sugar industry in Egypt. It is threatened by several insect pests among most important of them is the beet fly Pegomyia mixta. This work deals with the biological control of this insect using four entomopathogenic nematodes (EPNs). The nematodes included Steinernema carpocapsae S2, Steinernema feltiae, Heterorhabditis bacteriophora (HB1-3) and Heterorhabditis bacteriophora S1. Daily mortality of larvae and pupae of P. mixta were recorded after treatment with serial concentrations (500, 1000, 2000 and 4000 infective juveniles (IJs)/ml) of each of four studied EPNs. In the laboratory all tested nematodes killed the larvae inside their mines in the sugar beet leaves and developed in their bodies in different extends. They also killed the insect pupae in the soil and developed in their bodies. Young larvae were more susceptible than old ones. New pupae were more susceptible than old ones. In the field a single spray of S. feltiae or H. bacteriophora caused 81.3 or 75.9% reduction in the larval population of the in sugar beet leaves.     

Molecular polymorphisms in Palestinian Figs (Ficus carica L.) as revealed by Random Amplified Polymorphic DNA (RAPD)

The genetic diversity of 13 local Palestinian fig genotypes was investigated using RAPD markers. Among the 30 tested primers, 28 revealed various banding patterns and 2 generated no polymorphic bands. In addition, 13 primers (46.4%) produced good amplification products with high intensity and pattern stability. A total of 94 DNA fragments (loci), separated by electrophoresis on agarose gel were detected, ranging in size from 190 to 1300 bp. Of these fragments, 72 (76.6%) were polymorphic and 22 (23.4%) were monomorphic. A minimum of three and a maximum of eight DNA fragments were obtained using (OPH-02 and OPT-10) as well as (OPA-13, OPA-18 and OPY-07) primers respectively. The maximum percentage of polymorphic markers was 100.0 (Z-5, Z-12, and OPT-10) and the minimum was 60.0 (OPH-02). Primers OPY-07 and OPA-13 revealed high collective resolving power (Rp) values with 4.640 and 4.760 respectively and therefore, they were the most useful RAPD primers to assess the genetic diversity in the Palestinian figs. Genetic distance matrix showed an average distance range from 0.186 to 0.559 with a mean of 0.373. Thus, the cultivars tested in this study were characterized by large divergence at the DNA level. To our knowledge, this is the first report using RAPD marker to assess genetic diversity of Palestinian figs.     

Domain organization differences explain Bcr-Abl's preference for CrkL over CrkII

CrkL is a key signaling protein that mediates the leukemogenic activity of Bcr-Abl. CrkL is thought to adopt a structure that is similar to that of its CrkII homolog. The two proteins share high sequence identity and indistinguishable ligand binding preferences, yet they have distinct physiological roles. Here we show that the structures of CrkL and phosphorylated CrkL are markedly different than the corresponding structures of CrkII. As a result, the binding activities of the Src homology 2 and Src homology 3 domains in the two proteins are regulated in a distinct manner and to a different extent. The different structural architecture of CrkL and CrkII may account for their distinct functional roles. The data show that CrkL forms a constitutive complex with Abl, thus explaining the strong preference of Bcr-Abl for CrkL. The results also highlight how the structural organization of the modular domains in adaptor proteins can control signaling outcome.     

Morphology and infraciliature of two new marine ciliates, Paracyrtophoron tropicum nov. gen., nov. spec. and Aegyria rostellum nov. spec. (Ciliophora, Cyrtophorida), isolated from tropical waters in southern China

The morphology and infraciliature of two new marine cyrtophorid ciliates, Paracyrtophoron tropicum nov. gen., nov. spec. and Aegyria rostellum nov. spec., isolated from tropical waters in southern China, were investigated using live observation and protargol impregnation methods. Paracyrtophoron nov. gen. differs from the closely related Cyrtophoron by lack of fragment kinety at anterior ends of right somatic kineties and thigmotactic cilia in posterior portion of ventral surface, while from the well-defined Chlamydodon by lack of the cross-striped band around the periphery of the somatic field. Paracyrtophoron tropicum nov. spec., the type of the new genus, can be recognized by the combination of the following characters: cell size about 150-175×70-90μm in vivo; elliptical to kidney-shaped in outline, dorsoventrally flattened about 2.5:1; conspicuous cortical granules; one canal-like depression extending from postoral area to subcaudal region of cell; ca. 90 somatic kineties; 12-16 nematodesmal rods; one or two terminal fragments on dorsal side. Aegyria rostellum is characterized by the following features: size about 90-150×40-70μm in vivo, triangular or ear-shaped body with broad anterior end, having a rostriform structure and pigment spots, 56-63 somatic kineties, one preoral kinety, three or four circumoral kineties, and 32-42 nematodesmal rods. Based on previous and current studies, the definition for the genus Aegyria is updated: body dorsoventrally flattened; oral ciliature consisting of one preoral and several circumoral kineties; podite located in posterior ventral region and surrounded by somatic kineties; no obvious gap between right and left somatic kineties; postoral and left somatic kineties progressively shortened posteriorly from right to left. Additionally, two new combinations were proposed.     

Developing species-specific primers to identify Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia

This work aimed to determine the inter- and intra-specific variations in populations of Bulinus truncatus and Bulinus beccari, the intermediate hosts of Schistosoma haematobium in Saudi Arabia, and to develop species-specific primers to identify these snails as a first step in the development of multiplex PCR for simultaneously identifying the snails and diagnosing its infections in a single step. Two populations of B. truncatus were collected from Asser and Bisha (A and B), and two B. beccari populations were collected from Mahial Asser and Merba (C and D). The snails' genomic DNA was extracted and amplified using 5 different primers. The primers displayed variable intra- and inter-specific differences across the populations. The largest RAPD-PCR fragments were cloned into a vector as a preparatory step for sequencing. Similarity searches for the sequenced cloned inserts revealed no similar sequences in the GenBank database or its associated databases. Specific primers used to target the B. truncatus and B. beccari genomes were designed using the Gene Runner program and based on the DNA sequences obtained from RAPD fragment sequence analyses. Using these primers for specific PCRs resulted in expected single-band PCR products of 536 bp for B. beccari and 478 bp for B. truncatus. These results will be helpful for simultaneously identifying B. truncatus and B. beccari snails and diagnosing S. haematobium infections within the snails using single step multiplex PCR.