Roda Al-Thani

Characterization of Tunisian Bacillus thuringiensis strains with abundance of kurstaki subspecies harbouring insecticidal activities against the lepidopteran insect Ephestia kuehniella.

The study of 257 crystal-producing Bacillus thuringiensis isolates from bioinsecticide free soil samples collected from different sites in Tunisia, was performed by PCR amplification, using six primer pairs specific for cry1, cry2, cry3, cry4, and vip3A genes, by the investigation of strain plasmid pattern, crystal morphology and delta-endotoxin content and by the assessment of insecticidal activities against the lepidopteran insect Ephestia kuehniella. Based on plasmid pattern study, 11 representative strains of the different classes were subjected to morphological and molecular analyses. The comparison of the PFGE fingerprints confirmed the heterogeneity of these strains. B. thuringiensiskurstaki strains, harbouring at the same time the genes cry1A, cry2, cry1Ia, and vip3A, were the most abundant (65.4%). 33.34% of the new isolates showed particular delta-endotoxin profiles but no PCR products with the used primer sets. B. thuringiensisisraelensis was shown to be also very rare among the Tunisian B. thuringiensis isolates diversity. These findings could have considerable impacts for the set up of new pest control biological agents.

Community Structure and Activity of a Highly Dynamic and Nutrient-Limited Hypersaline Microbial Mat in Um Alhool Sabkha, Qatar

The Um Alhool area in Qatar is a dynamic evaporative ecosystem that receives seawater from below as it is surrounded by sand dunes. We investigated the chemical composition, the microbial activity and biodiversity of the four main layers (L1–L4) in the photosynthetic mats. Chlorophyll a (Chl a) concentration and distribution (measured by HPLC and hyperspectral imaging, respectively), the phycocyanin distribution (scanned with hyperspectral imaging), oxygenic photosynthesis (determined by microsensor), and the abundance of photosynthetic microorganisms (from 16S and 18S rRNA sequencing) decreased with depth in the euphotic layer (L1). Incident irradiance exponentially attenuated in the same zone reaching 1% at 1.7-mm depth. Proteobacteria dominated all layers of the mat (24%–42% of the identified bacteria). Anoxygenic photosynthetic bacteria (dominated by Chloroflexus) were most abundant in the third red layer of the mat (L3), evidenced by the spectral signature of Bacteriochlorophyll as well as by sequencing. The deep, black layer (L4) was dominated by sulfate reducing bacteria belonging to the Deltaproteobacteria, which were responsible for high sulfate reduction rates (measured using 35S tracer). Members of Halobacteria were the dominant Archaea in all layers of the mat (92%–97%), whereas Nematodes were the main Eukaryotes (up to 87%). Primary productivity rates of Um Alhool mat were similar to those of other hypersaline microbial mats. However, sulfate reduction rates were relatively low, indicating that oxygenic respiration contributes more to organic material degradation than sulfate reduction, because of bioturbation. Although Um Alhool hypersaline mat is a nutrient-limited ecosystem, it is interestingly dynamic and phylogenetically highly diverse. All its components work in a highly efficient and synchronized way to compensate for the lack of nutrient supply provided during regular inundation periods.

Isolation, biochemical and molecular characterization of 2-chlorophenol-degrading Bacillus isolates

Pure cultures of 2-chlorophenol degrading bacteria were isolated from a natural enrichment that may be adapted to chlorophenols in the industrial zone at Umm-Saied city (Qatar). The bacteria were identified by 16S rDNA analysis, using PCR with universal primers. Comparative analysis of the 16S rDNA sequence (~ 550 bp) in the GenBank database revealed that these bacteria are related to the genus Bacillus. Molecular heterogeneity among 2-chlorophenol-degrading bacteria was investigated using REP-PCR chromosomal fingerprinting and correlated with antibiotic profile analysis. REP-PCR results strongly confirmed that the bacterial isolates from different Qatari soils produced different fingerprinting patterns. The distribution of phenol hydroxylase catabolic gene among examined isolates revealed that three isolates out of six yielded positive PCR products. Degradation of 2-chlorophenol was studied using theses cultures in liquid medium under aerobic conditions, at initial concentrations of 0.25 – 2.5 mM 2-chlorophenol. Undegraded 2-chlorophenol was quantified by high-performance liquid chromatography (HPLC). Degradation rates by isolates could be determined at concentrations up to 1.5 mM. However, higher concentrations of 2-chlorophenol (2.5 mM) were inhibitory to cell growth.

Cyanobacteria, Oil – and Cyanofuel?

Global warming, the global carbon cycle, and current and future global trading in carbon credits (the carbon market), are beginning to dictate radical changes in human behaviour. In this context, the cyanobacteria figure prominently. Why? First, vast populations of ancient cyanobacteria and other microalgae are credited with the formation of Earth’s oil deposits. Second, extant populations of cyanobacteria, most conspicuously marine picoplankton (Chaps. 5 and 13) contribute significantly to the fixation of atmospheric carbon through their photosynthesis. Third, spills from the commercial trafficking of oil often accumulate in coastal regions where cyanobacterial mats are prevalent (Chap. 4), and this led to the examination of how these microorganisms participate in mitigation of the effects of oil pollution. Fourth, cyanobacteria may be a viable source of biofuel. As such, the rise of cyanobacteria, cyanobacteria and oil pollution, and cyanobacteria as a source of biofuel (cyanofuel) can be equated, respectfully, with Earth’s past, present and future. In this chapter we emphasize connections between all three through consideration of cyanobacterial physiology, ecology and molecular biology. We wish to emphasize the persistence of cyanobacteria through geological time and their tenacious hold on carbon.

Patulin and patulin producing Penicillium spp. occurrence in apples and apple-based products including baby food

INTRODUCTION Patulin has raised the international attention because of its health risk. In fact, it has mutagenic, neurotoxic, immunotoxic, genotoxic and gastrointestinal effects in animals. In the present work, patulin and patulin-producing Penicillium spp. in apple and apple-based products marketed in Qatar were analysed. METHODOLOGY Sampling was carried out using apple fruits and apple-based products. Fungi were isolated from undamaged apples, apple juice and baby apple food. DNA extraction was carried out with DNeasy Plant Mini Kit (QIAGEN, Valencia, USA). The molecular identification of fungal isolates was carried out using ITS1-ITS4 PCR. PCR products were sequenced and blasted. Patulin was extracted and analyzed by LC/MS/MS, then quantified using Agilent 1290UHPLC coupled to 6460 triple quadruple mass spectrometer. RESULTS Forty-five samples of undamaged fresh apple fruits, apple juice and apple-based baby food products sold in different markets in Qatar were surveyed for both fungal and patulin contamination using Liquid Chromatography Tandem Mass Spectrometery (LC/MS/MS). Twenty-five Penicillium spp. isolates were selected, including 23 P. expansum and one isolate each of P. brevicompactum and P. commune. All the tested Penicillium spp. isolates produced patulin in vitro (from 40 to 100 μg/g on Malt Yeast Extract agar medium). Patulin was detected in 100% of apple juice samples at levels ranging from 5.27 to 82.21 µg/kg. Only 5 samples contained patulin levels higher than European Union recommended limit (50 µg/kg). The average patulin contamination was 30.67 µg/kg and 10.92 µg/kg in baby apple juice and in baby apple compote, respectively.

Application of Low-Fermenting Yeast Lachancea thermotolerans for the Control of Toxigenic Fungi Aspergillus parasiticus, Penicillium verrucosum and Fusarium graminearum and Their Mycotoxins

Mycotoxins are important contaminants of food and feed. In this study, low fermenting yeast (Lachancea thermotolerans) and its derivatives were applied against toxigenic fungi and their mycotoxins. A. parasiticus, P. verrucosum and F. graminearum and their mycotoxins were exposed to yeast volatile organic compounds (VOCs) and cells, respectively. VOCs reduced significantly the fungal growth (up to 48%) and the sporulation and mycotoxin synthesis (up to 96%). Very interestingly, it was shown that even 7 yeast colonies reduced Fusarium’s growth and the synthesis of its mycotoxin, deoxynivalenol (DON). Moreover, decreasing yeast nutrient concentrations did not affect the inhibition of fungal growth, but reduced DON synthesis. In addition, inactivated yeast cells were able to remove up to 82% of the ochratoxin A (OTA). As an application of these findings, the potentialities of the VOCs to protect tomatoes inoculated with F. oxysporum was explored and showed that while in the presence of VOCs, no growth was observed of F. oxysporum on the inoculated surface areas of tomatoes, in the absence of VOCs, F. oxysporum infection reached up to 76% of the tomatoes’ surface areas. These results demonstrate that the application of yeasts and their derivatives in the agriculture and food industry might be considered as a very promising and safe biocontrol approach for food contamination.

The Replacement of five Consecutive Amino Acids in the Cyt1A Protein of Bacillus thuringiensis Enhances its Cytotoxic Activity against Lung Epithelial Cancer Cells

Cyt1A protein is a cytolytic protein encoded by the cyt gene of Bacillus thuringiensis subsp. israelensis (Bti) as part of the parasporal crystal proteins produced during the sporulation. Cyt1A protein is unique compared to the other endotoxins present in these parasporal crystals. Unlike δ-endotoxins, Cyt1A protein does not require receptors to bind to the target cell and activate the toxicity. It has the ability to affect a broad range of cell types and organisms, due to this characteristic. Cyt1A has been recognized to not only target the insect cells directly, but also recruit other endotoxins by acting as receptors. Due to these mode of actions, Cyt1A has been studied for its cytolytic activity against human cancer cell lines, although not extensively. In this study, we report a novel Cyt1A protein produced by a Bti strain QBT229 isolated from Qatar. When tested for its cytotoxicity against lung cancer cells, this local strain showed considerably higher activity compared to that of the reference Bti and other strains tested. The possible reasons for such enhanced activity were explored at the gene and protein levels. It was evidenced that five consecutive amino acid replacements in the β8 sheet of the Cyt1A protein enhanced the cytotoxicity against the lung epithelial cancer cells. Such novel Cyt1A protein with high cytotoxicity against lung cancer cells has been characterized and reported through this study.

Solutes in native plants in the Arabian Gulf region and the role of microorganisms: future research

Aims One of the outstanding challenges facing humankind is increasing crop production under various types of severe environmental conditions. Many measures have been taken to adopt molecular and biotechnological approaches that lead to the development of transgenic plants able to deal with such harsh and polluted environments. However, such solutions could be very expensive and require considerable efforts and time to achieve these objectives. The main objective of this review is to discuss the new biological solutions that have emerged in the last decade, as environmentally friendly approaches, perhaps to support and/or replace the present efforts. These solutions based on plant–microbe interactions could be a lifeline and promising alternative strategy to create plants with a high resistance to the extreme environments. Methods During the last two decades research projects have been conducted to study the ecology, identify the features, and the ecophysiology of native plants and the associated microorganisms in the Arabian Gulf region and particularly in Qatar. Many physiological and biochemical parameters have been determined, including organic solutes (amino acids like proline, glycinebetaine, soluble sugars, etc.), photosynthetic pigments, organic acids and inorganic ions especially heavy metals, along with the physical and chemical properties of the soil in various locations of the State of Qatar. Also, the microorganisms adjacent to and associated with these native plants were identified to elucidate the possible roles in the soil biota in supporting these plants against extreme environmental conditions. Important Findings Investigations of native plants in the Arabian Gulf states during the last decade have shown that wild plants exhibit different abilities to accumulate organic solutes to cope with the harsh natural environments. Pollution is a major factor stressing wildlife in this region due to the expansion of urban sectors and industrial activities of oil and gas. Compatible osmolytes, like proline, accumulate in wild plants in response to severe environmental conditions and heavy metal contaminated soil. Accumulation of these solutes in plant tissues could provide some level of adaptation and resistance against all these types of environmental stresses. We present some promising efforts in the Arabian Gulf region to remediate desert soil and water polluted with heavy metals and petroleum hydrocarbons. Substantial evidence is introduced about the roles of microorganisms associated with wild plants in natural habitats, such association may help them cope with the extreme stresses. Possible mechanisms adopted by microorganisms in alleviating the harsh abiotic stresses facing the wild life are discussed, one of which is the promotion of biosynthesis and transport of organic solutes to the plants. Also, the main possibilities of the origin of activities of the accumulation of compatible organic solutes are suggested and the objectives of the future research are discussed.

Diversity of Bacillus thuringiensis Strains From Qatar as Shown by Crystal Morphology, d-Endotoxins and Cry Gene Content

Bacillus thuringiensis (Bt) based insecticidal formulations have been recognized as one of the most successful, environmentally safe and sustainable method of controlling insect pests. Research teams worldwide are in search of Bt diversity giving more choices of bio-insecticides and alternatives to address insect resistance. In fact, there are many unexplored ecologies that could harbor novel Bt strains. This study is the first initiative to explore Bt strain diversity in Qatar. A collection of 700 Bt isolates was constructed. Scanning electron microscopy of Bt crystals showed different crystal forms, with a high abundance of spherical crystals compared to the bipyramidal ones. Among the spherical crystals, four different morphologies were observed. The δ-endotoxin content of parasporal crystals from each Bt isolate revealed that there are 16 different protein profiles among the isolates of the collection. On the other hand, plasmid pattern analysis showed seven different plasmid profiles. Their insecticidal activity was predicted by exploring the δ-endotoxin coding genes and conducting qualitative insecticidal bioassays. 19 smooth spherical crystal producing isolates have been identified that could be possible candidates for endotoxin production targeting Dipteran insects. Another group of 259 isolates producing bipyramidal and cuboidal crystals could target Lepidopteran and Coleopteran insects. The remaining 422 isolates have novel profiles. In conclusion, Qatari soil ecology provides a good collection and diversity of Bt isolates. In addition to strains harboring genes encoding common endotoxins, the majority are different and very promising for the search of novel insecticidal endotoxins.