Aerobic and facultative anaerobic bacteria from gut of red palm weevil (Rhynchophorus ferrugineus)

Red palm weevil (RPW), Rhynchophorus ferrugineus Oliver is one of the insects that attack date palm trees directly and cause its death. This infection poses serious economical consequences in SaudiArabia and other gulf countries. RPW assimilates the components of palm tree tissues. Significant amount of microbiota in the gut of RPW may contribute to success of its pathogenesis. This studyexplored the nature of microbiota in RPW by culture-dependent and molecular-based techniques using PCR and sequencing analysis. Larvae of RPW were fed on an artificial diet and were fixed beforeopening its gut for microbial cultivation on enrichment media. Pure cultures were obtained after incubating the plates at different atmospheric conditions (aerobic, and strictly anaerobic). The majorityof isolated microbiota observed were aerobes and facultative anaerobes (Bacillus sp., Salmonella sp., Enterococcus sp., and Xanthomonas sp.). These qualitative differences of bacteria, suggest the presence of a complex ecosystem in the gut of RPW. Subsequently, bacterial DNAs were extracted from pure cultures for definitive molecular identifications. Hot start-touchdown PCR was performed toamplify regions within 16S rDNA. Amplicons were cloned into the TOPO-TA vector for sequencing. The data reveal to some extent that aerobic and facultative anaerobic bacteria are more distributed in the gut of RPW

New insights into Plagiogrammaceae (Bacillariophyta) based on multigene phylogenies and morphological characteristics with the description of a new genus and three new species.

Plagiogrammaceae, a poorly described family of diatoms, are common inhabitants of the shallow marine littoral zone, occurring either in the sediments or as epiphytes. Previous molecular phylogenies of the Plagiogrammaceae were inferred but included only up to six genera: Plagiogramma, Dimeregramma, Neofragilaria, Talaroneis, Psammogramma and Psammoneis. In this paper, we describe a new plagiogrammoid genus, Orizaformis, obtained from Bohai Sea (China) and present molecular phylogenies of the family based on three and four genes (nuclear-encoded large and small subunit ribosomal RNAs and chloroplast-encoded rbcL and psbC). Also included in the new phylogenies is Glyphodesmis. The phylogenies suggest that the Plagiogrammaceae is composed of two major clades: one consisting of Talaroneis, Orizaformis and Psammoneis, and the second of Glyphodesmis, Psammogramma, Neofragilaria, Dimeregramma and Plagiogramma. In addition, we describe three new species within established genera: Psammoneis obaidii, which was collected from the Red Sea, Saudi Arabia; and Neofragilaria stilus and Talaroneis biacutifrons from the Mozambique Channel, Indian Ocean, and illustrate two new combination taxa: Neofragilaria anomala and Neofragilaria lineata. Our observations suggest that the biodiversity of the family is strongly needed to be researched, and the phylogenetic analyses provide a useful framework for future studies of Plagiogrammaceae

Antilisterial Effects of Gravinol-S Grape Seed Extract at Low Levels in Aqueous Media and Its Potential Application as a Produce Wash

Grape seed extract (GSE) is a rich source of proanthocyanidins, a class of natural antioxidants reported to have wide-ranging bioactivity as anti-inflammatory, anticarcinogenic, and antimicrobial agents. The ability of GSE to rapidly inactivate Listeria monocytogenes in vitro and the generally recognized as safe status of GSE make this extract an attractive candidate for control of Listeria in or on foods. Previously, GSE has been used at relatively high concentrations (1%) in complex food matrices and in combination with other antimicrobials. We sought to characterize the antilisterial effects of a commercial GSE preparation (Gravinol-S) alone at much lower concentrations (0.00015 to 0.125%) in aqueous solution and to test its possible use as an antimicrobial wash for fresh produce surfaces. Based on broth microdilution tests, the MICs of GSE against L. monocytogenes Scott A and Listeria innocua ATCC 33090 were as low as 50 and 78 mg ml21, respectively. GSE was evaluated in 0.85% saline against live cells of L. innocua via flow cytometry, using propidium iodide as a probe for membrane integrity. At sub-MICs and after only 2 min of exposure, treatment with GSE caused rapid permeabilization and clumping of L. innocua, results that we confirmed for L. monocytogenes using fluorescence microscopy and Live/Dead staining. At higher concentrations (0.125%), GSE reduced viable cell counts for L. monocytogenes by approximately 2 log units within 2 min on tomato surfaces. These results suggest the potential for GSE as a natural control of Listeria spp. on low-complexity foods such as tomatoes.This article is from Journal of Food Protection 73(2): 266-273. Posted with permission.</p

تقييم طرق مسح نشاط أنزيم السيليوليز خارج الخلوي في بعض الفطريات الخيطية

Twenty-eight fungal species belonging to fifteen genera were isolated from Saudi Arabian soil and were screened for the production of extracellular cellulase enzymes. Cleared-zone technique, cellulose azure method and agar diffusion test for rapid screening were used. The cleared-zone technique was found to be much less reliable as compared to other methods. Trichoderma viride, T. pseudokoningii and Aspergillus fumigatus were the best producers of cellulase enzyme. The data obtained also suggest that caution is needed using these methods. The disadvantages of using one method for screening, which has been a common practise up to now, are discussed

MicroRNA-375 and MicroRNA-221: Potential Noncoding RNAs Associated with Antiproliferative Activity of Benzyl Isothiocyanate in Pancreatic Cancer

The deregulated presence or absence of microRNAs (miRNAs) might play an important role in molecular pathways leading to neoplastic transformation. At present, it is also thought that the approaches to interfere miRNA functions should be helpful for developing novel therapeutic opportunities for human cancer. In this study, we provide evidence that the anticancer agent benzyl isothiocyanate (BITC) has the ability to modulate the level of miRNAs such as miR-221 and miR-375, known to be abnormally expressed in pancreatic cancer patients. Interestingly, ectopic expression of miR-375 or the enforced silencing of miR-221 in cultured pancreatic cancer cells attenuates cell viability and sensitizes antiproliferative action of BITC. We also show that the expression of putative tumor suppressor miR-375 is more abundant in nonpathological mice pancreata than those with KrasG12D-driven pancreatic intraepithelial neoplasia (PanIN). To the contrary, the expression of oncogenic miR-221 is significantly elevated in the mouse pancreas with PanIN lesions. Although miR-375 has been shown to be aberrantly expressed in pancreatic cancer patients, there has not been a comprehensive study to investigate the molecular pathways targeted by this miRNA in pancreatic cancer cells. Further analysis by gene expression microarray revealed that IGFBP5 and CAV-1, potential biomarkers of pancreatic cancer, were significantly downregulated in cells transfected with miR-375. Correlatively, elevated expression of IGFBP5 and CAV-1 was evident in the mouse pancreas with preneoplastic lesions in which the expression of miR-375 wanes. Taken together, our findings suggest that anticancer agent BITC might target the expression of miR-221 and miR-375 to switch hyperproliferative pancreatic cancer cells to a hypoproliferative state

Efficacy of neutral and negatively charged liposome-loaded gentamicin on planktonic bacteria and biofilm communities

Moayad Alhariri,1 Majed A Majrashi,2 Ali H Bahkali,3 Faisal S Almajed,4 Ali O Azghani,5 Mohammad A Khiyami,2 Essam J Alyamani,2 Sameera M Aljohani,6 Majed A Halwani1 1Nanomedicine Department, King Abdullah International Medical Research Center (KAIMRC), King Saud bin Abdulaziz University for Health Sciences, 2National Centre for Biotechnology, Life Sciences and Environment Research Institute, King Abdulaziz City for Science and Technology (KACST), 3Botany and Microbiology Department, College of Science, King Saud University, 4Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud bin Abdulaziz University for Health Sciences, King Abdullah International Medical Research Center, Riyadh, Saudi Arabia; 5Department of Biology, The University of Texas at Tyler, Tyler, TX, USA; 6College of Science and Health Professions, King Saud bin Abdulaziz University for Health Sciences, King Abdullah International Medical Research Center, Riyadh, Saudi Arabia Abstract: We investigated the efficacy of liposomal gentamicin formulations of different surface charges against Pseudomonas aeruginosa and Klebsiella oxytoca. The liposomal gentamicin formulations were prepared by the dehydration&ndash;rehydration method, and their sizes and zeta potential were measured. Gentamicin encapsulation efficiency inside the liposomal formulations was determined by microbiologic assay, and stability of the formulations in biologic fluid was evaluated for a period of 48 h. The minimum inhibitory concentration and the minimum bactericidal concentration were determined, and the in vitro time kill studies of the free form of gentamicin and liposomal gentamicin formulations were performed. The activities of liposomal gentamicin in preventing and reducing biofilm-forming P. aeruginosa and K. oxytoca were compared to those of free antibiotic. The sizes of the liposomal formulations ranged from 625 to 806.6 nm in diameter, with the zeta potential ranging from &ndash;0.22 to &ndash;31.7 mV. Gentamicin encapsulation efficiency inside the liposomal formulation ranged from 1.8% to 43.6%. The liposomes retained &gt;60% of their gentamicin content during the 48 h time period. The minimum inhibitory concentration of neutral formulation was lower than that of free gentamicin (0.25 versus 1 mg/L for P. aeruginosa and 0.5 versus 1 mg/L for K. oxytoca). The negatively charged formulation exhibited the same bacteriostatic concentration as that of free gentamicin. The minimum bactericidal concentration of neutral liposomes on planktonic bacterial culture was twofold lower than that of free gentamicin, whereas the negatively charged formulations were comparable to free gentamicin. The killing time curve values for the neutral negatively charged formulation against planktonic P. aeruginosa and K. oxytoca were better than those of free gentamicin. Furthermore, liposomal formulations prevent the biofilm-formation ability of these strains better than free gentamicin. In summary, liposomal formulations could be an effective lipid nanoparticle to combat acute infections where planktonic bacteria are predominant. Keywords: drug delivery, stability, antibacterial activity, biofil