Alex Sivan

New perspectives in plastic biodegradation.

During the past 50 years new plastic materials, in various applications, have gradually replaced the traditional metal, wood, leather materials. Ironically, the most preferred property of plastics--durability--exerts also the major environmental threat. Recycling has practically failed to provide a safe solution for disposal of plastic waste (only 5% out of 1 trillion plastic bags, annually produced in the US alone, are being recycled). Since the most utilized plastic is polyethylene (PE; ca. 140 million tons/year), any reduction in the accumulation of PE waste alone would have a major impact on the overall reduction of the plastic waste in the environment. Since PE is considered to be practically inert, efforts were made to isolate unique microorganisms capable of utilizing synthetic polymers. Recent data showed that biodegradation of plastic waste with selected microbial strains became a viable solution.

Integrated control of Fusarium crown and root rot of tomato with Trichoderma harzianum in combination with methyl bromide or soil solarization.

Abstract Combination of Trichoderma harzianum with soil solarization or with a reduced dose of methyl bromide, under field conditions, resulted in significant ( p = 0.05) disease control of fusarium crown and root rot of tomato induced by Fusarium oxysporum f. sp. radicis-lycopersici (FORL). T. harzianum or soil solarization applied alone were ineffective in protecting the tomato plants from crown and root rot. Soil fumigation with a sub-lethal dose of methyl bromide (300 kg ha −1 ) provided the same degree of protection as the recommended dose (750 kg ha −1 ). Maximum disease control (48%) was obtained with a combination of T. harzianum and the reduced dose of the fumigant. Treatments that resulted in disease control also elicited a significant yield increase ( p = 0.05). The highest yield improvement (105% over the control) was recorded in plots where the antagonist had been applied in combination with soil solarization. In plots not treated with T. harzianum , the rhizosphere soil contained low levels of Trichoderma spp. [10 2 -10 3 colony-forming units (c.f.u.) g −1 rhizosphere soil], and tomato root segments showed no evidence of colonization by the antagonist. High levels of Trichoderma (10 4 −10 5 c.f.u. g −1 ) were recovered from rhizosphere soil or crown segments of plants treated with the antagonist in combination with methyl bromide or soil solarization. Conversely, high population levels of Fusarium spp. (2 × 10 5 − 3 × 10 7 c.f.u. g −1 ) were recorded in rhizosphere soil or on crown segments sampled from all treatments. The combination of T. harzianum with methyl bromide (300 kg ha −1 ) or soil solarization reduced the final colonization level of Fusarium spp. on the crown surface by 76 or 94%, respectively. This study reveals that combination of T. harzianum with a sub-lethal dose of methyl bromide or with soil solarization is effective in controlling FORL in tomato.

Global distribution and diversity of coral‐associated Archaea and their possible role in the coral holobiont nitrogen cycle

Diversity, distribution and genetic comparison of Archaea associated with the surface mucus of corals from three genera, namely Acanthastrea sp., Favia sp. and Fungia sp., from the Gulf of Eilat, Israel and from Heron Island, Australia were studied. Sequencing of the 16S rRNA gene of the coral-associated Archaea revealed dominance of Crenarchaeota (79%, on average). In this phylum, 87% of the sequences were similar (>or= 97%) to the Thermoprotei, with 76% of these being similar (>or= 97%) to the ammonium oxidizer, Nitrosopumilus maritimus. Most of the coral-associated euryarchaeotal sequences (69%) were related to marine group II, while other euryarchaeotal clades were found to be related to anaerobic methanotrophs (8%), anaerobic nitrate reducers (i.e. denitrification, 15%) and marine group III (8%). Most of the crenarchaeotal and euryarchaeotal coral-associated 16S rRNA gene sequences from Heron Island (61%) and from the Gulf of Eilat (71%) were closely related (>or= 97%) to sequences previously derived from corals from the Virgin Islands. Analysis of archaeal amoA sequences obtained from the fungiid coral, Fungia granulosa, divided into three clades, all related to archaeal sequences previously obtained from the marine environment. These sequences were distantly related to amoA sequences previously found in association with other coral species. Preliminary experiments suggest that there is active oxidation of ammonia to nitrite in the mucus of F. granulosa. Thus, coral-associated Archaea may contribute to nitrogen recycling in the holobiont, presumably by acting as a nutritional sink for excess ammonium trapped in the mucus layer, through nitrification and denitrification processes.

Accelerated biodegradation of cement by sulfur-oxidizing bacteria as a bioassay for evaluating immobilization of low-level radioactive waste

ABSTRACT Disposal of low-level radioactive waste by immobilization in cement is being evaluated worldwide. The stability of cement in the environment may be impaired by sulfur-oxidizing bacteria that corrode the cement by producing sulfuric acid. Since this process is so slow that it is not possible to perform studies of the degradation kinetics and to test cement mixtures with increased durability, procedures that accelerate the biodegradation are required. Semicontinuous cultures of Halothiobacillus neapolitanus and Thiomonas intermedia containing thiosulfate as the sole energy source were employed to accelerate the biodegradation of cement samples. This resulted in a weight loss of up to 16% after 39 days, compared with a weight loss of 0.8% in noninoculated controls. Scanning electron microscopy of the degraded cement samples revealed deep cracks, which could be associated with the formation of low-density corrosion products in the interior of the cement. Accelerated biodegradation was also evident from the leaching rates of Ca2+ and Si2+, the major constituents of the cement matrix, and Ca exhibited the highest rate (up to 20 times greater than the control rate) due to the reaction between free lime and the biogenic sulfuric acid. Leaching of Sr2+ and Cs+, which were added to the cement to simulate immobilization of the corresponding radioisotopes, was also monitored. In contrast to the linear leaching kinetics of calcium, silicon, and strontium, the leaching pattern of cesium produced a saturation curve similar to the control curve. Presumably, the leaching of cesium is governed by the diffusion process, whereas the leaching kinetics of the other three ions seems to governed by dissolution of the cement.

Biofilm production by clinical strains of Acinetobacter baumannii isolated from patients hospitalized in two tertiary care hospitals

Microbial biofilms are considered as virulence factors. During the present study, 34 clinical strains of Acinetobacter baumannii, isolated from patients hospitalized in two tertiary care hospitals, were examined for biofilm formation. These strains showed high variability in biofilm formation. Furthermore, no relation could be found between the ability of biofilm production and molecular type, carbapenem resistance, site of isolation of the clinical strains of A. baumannii and disease severity. Interestingly, in two cases an increase in biofilm formation could be detected in A. baumannii isolates cultured from the same patient upon prolonged hospitalization.

Antibacterial Activity of Pseudoalteromonas in the Coral Holobiont

Corals harbor diverse and abundant prokaryotic populations. Bacterial communities residing in the coral mucus layer may be either pathogenic or symbiotic. Some species may produce antibiotics as a method of controlling populations of competing microbial species. The present study characterizes cultivable Pseudoalteromonas sp. isolated from the mucus layer of different coral species from the northern Gulf of Eilat, Red Sea, Israel. Six mucus-associated Pseudoalteromonas spp. obtained from different coral species were screened for antibacterial activity against 23 tester strains. Five of the six Pseudoalteromonas strains demonstrated extracellular antibacterial activity against Gram-positive—but not Gram-negative—tester strains. Active substances secreted into the cell-free supernatant are heat-tolerant and inhibit growth of Bacillus cereus, Staphylococcus aureus, and of ten endogenous Gram-positive marine bacteria isolated from corals. The Pseudoalteromonas spp. isolated from Red sea corals aligned in a phylogenetic tree with previously isolated Pseudoalteromonas spp. of marine origin that demonstrated antimicrobial activity. These results suggest that coral mucus-associated Pseudoalteromonas may play a protective role in the coral holobionts defense against potential Gram-positive coral pathogens.

Effect of proteases on biofilm formation of the plastic-degrading actinomycete Rhodococcus ruber C208.

In most habitats, the vast majority of microbial populations form biofilms on solid surfaces, whether natural or artificial. These biofilms provide either increased physical support and/or a source of nutrients. Further modifications and development of biofilms are regulated by signal molecules secreted by the cells. Because synthetic polymers are not soluble in aqueous solutions, biofilm-producing bacteria may biodegrade such materials more efficiently than planktonic strains. Bacterial biofilms comprise bacterial cells embedded in self-secreted extracellular polymeric substances (EPS). Revealing the roles of each component of the EPS will enable further insight into biofilm development and the EPS structure-function relationship. A strain of Rhodococcus ruber (C208) displayed high hydrophobicity and formed a dense biofilm on the surface of polyethylene films while utilizing the polyolefin as carbon and energy sources. This study investigated the effects of several proteases on C208 biofilm formation and stability. The proteolysis of C208 biofilm gave conflicting results. Trypsin significantly reduced biofilm formation, and the resultant biofilm appeared monolayered. In contrast, proteinase K enhanced biofilm formation, which was robust and multilayered. Presumably, proteinase K degraded self-secreted proteases or quorum-sensing peptides, which may be involved in biofilm detachment processes, leading to a multilayered, nondispersed biofilm.

Stramenopile Microorganisms Associated with the Massive Coral Favia sp.

ABSTRACT. The surfaces of massive corals of the genus Favia from Eilat, Red Sea, and from Heron Island, Great Barrier Reef, are covered by a layer of eukaryotic microorganisms. These microorganisms are embedded in the coral mucus and tissue. In the Gulf of Eilat, the prevalence of corals covered by patches of eukaryotic microorganisms was positively correlated with a decrease in water temperatures (from 25–28 °C in the summer to 20–23 °C in winter). Comparisons carried out using transmission and scanning electron microscopy showed morphological similarities between the microorganisms from the two geographically distant reefs. The microorganisms found on and in the tissues were approximately 5–15 μm in diameter, surrounded by scales in their cell wall, contained a nucleus, and included unique auto‐florescent coccoid bodies of approximately 1 μm. Such morphological characters suggested that these microorganisms are stramenopile protists and in particular thraustochytrids. Molecular analysis, carried out using specific primers for stramenopile 18S rRNA genes, revealed that 90% (111/123) of the clones in the gene libraries were from the Thraustochytriidae. The dominant genera in this family were Aplanochytrium sp., Thraustochytrium sp., and Labyrinthuloides sp. Ten stramenopile strains were isolated and cultured from the corals. Some strains showed ≥97% similarity to clones derived from libraries of mucus‐associated microorganisms retrieved directly from these corals. Fatty acid characterization of one of the prevalent strains revealed a high percentage of polyunsaturated fatty acids, including omega‐3. The possible association of these stramenopiles in the coral holobiont appeared to be a positive one.

Geographic Specific Coral-Associated Ammonia-Oxidizing Archaea in the Northern Gulf of Eilat (Red Sea)

Coral holobionts are densely populated with microorganisms that are essential for their well-being. Here we compared the diversity of the archaeal ammonia monooxygenase alpha subunit (amoA) gene from three coral genera, Acanthastrea sp., Favia sp., and Fungia granulosa, from the Gulf of Eilat, Red Sea. At 99% similarity, archaeal amoA from the three coral genera shared 71% of their cloned sequences, while the Favia and Acanthastrea presented a few genus-specific clones. In addition, the sequences retrieved in our samples displayed lower similarity to amoA sequences previously found in association with other coral species from different geographic regions. This finding suggests that the populations of ammonia-oxidizing archaea are less host-specific and more geographically dependent.

NOTE MOLECULAR ANALYSIS OF THE AHAS GENE OF PORPHYRIDIUM SP. (RHODOPHYTA) AND OF A MUTANT RESISTANT TO SULFOMETURON METHYL

Acetohydroxyacid synthase (AHAS) is the target enzyme of the sulfonylurea herbicides, and here we report the sequence of the gene from wild‐type and herbicide‐resistant Porphyridium sp. (Rhodophyta). The resistant mutant has a single residue substitution at a position known to confer herbicide resistance in E. coli and in plants. The rhodophyte gene is of cyanobacterial origin and distinct from the nuclear‐encoded chlorophyte gene, which may be of mitochondrial origin.