Detection of antibiotic-producing Actinobacteria in the sediment and water of Ma'in thermal springs (Jordan).

INTRODUCTION: Detection of new Actinobacteria is significant to discover new antibiotics because development of new antibiotics is connected to the characterization of novel bacterial taxa. This study has focused on the identification and isolation of antibiotic-producing Actinobacteria from the sediment and the water of Ma'in thermal springs (48-59°C) situated in the center area of Jordan. METHODS: Samples of sediment and water were transferred to glucose yeast malt agar medium and Actinobacteria were cultivated, isolated and identified according to scanning electron microscopy and 16S rRNA gene analysis. Antibacterial activities of the isolates were then tested against different test bacteria by agar well diffusion method. RESULTS: Three different species of Actinobacteria were isolated (M1-1, M2-2, M3-2) from sediment samples. Based on 16S rRNA gene analysis, isolate M1-1 was found to have only 90% identity percentage with Nocardiopsis sp., however, isolates M2-2 and M3-2 were found to be closely related Streptomyces sp. (97%) and Nocardioides luteus (99%), respectively. The antibacterial activity showed that strain M1-1 is active against P. aeruginosa ATCC 2785 (inhibition zone, 9 mm). Strain M2-2 was found to be active against S. aureus ATCC 29213 (12 mm), B. cereus ATCC 11778 (11 mm), and E. coli ATCC 25922 (9 mm). In respect to strain M3-2, it was found to be active against S. aureus ATCC 29213 (14 mm) and B. cereus ATCC 11778 (9 mm). There were no actinobacterial isolates obtained from water samples despite their significant diversity revealed by our previous metagenomic analysis, which showed the presence of 13 different species dominated by Arthrobacter (an Actinobacterium belonging to family Actinomycetales). CONCLUSION: There were 17 different Actinobacteria that could be detected in Ma'in thermal springs (13 unculturable species and 3 culturable species). The culturable Actinobacteria were found to have some antimicrobial activity. Further chemical analysis of the bioactive compounds is recommended.

Exploring the microbial diversity in Jordanian hot springs by comparative metagenomic analysis.

A culture-independent approach was utilized in this study to reveal the microbial diversity in Jordanian hot springs represented by Ma'in and Afra hot springs. Water samples from Ma'in and Afra hot springs were collected in June 2015. The in situ temperature of water samples range was 38-59°C and the pH range was 7.4-8.4. The metagenome was extracted and analyzed using the next generation technology (bTEFAP® ). A total of 314,310 sequences were parsed and 288,452 were then clustered. The sequences were predominated by bacteria (>84%) and the relative abundance of archaea in each sample was <1%. Eukaryotic microorganisms were detected but with varying abundances (0.6%-15%). Because most of the detected sequences were found to belong to the domain of bacteria (196,936 sequences out 288,452), the bacterial sequences were utilized for further microbial analyses. With respect to alpha and beta diversity, samples were rarefied to 30,000 sequences and bootstrapped at 10,000 sequences. The Shannon-Wiener Index curve plot reaches a plateau at approximately 3,000 sequences indicating that sequencing depth was sufficient to capture the full scope of microbial diversity. By examining the relative abundance of phyla detected in each sample, it appears that the biota of both Jordanian hot springs sampled are compositionally similar, with over 50% of the microbial community of each sample being comprised of the phylum Proteobacteria. The second most abundant phylum was the phylum Bacteroidetes which represents more than 13% in each sample. The phylum Firmicutes was also detected with a significant abundance. However, lower abundance of Deinococcus, Verrucomicrobia, Planctomycetes, and Chloroflexi was detected. A principal coordinate analysis plot was generated based upon the weighted UniFrac distance matrix. By utilizing Monte Carlo simulations, we were able to determine that there were no significant differences in the microbial diversity between each sample.

Microbiological analysis, antimicrobial activity, and heavy-metals content of Jordanian Ma'in hot-springs water.

Ma'in hot springs are known as sites of balneotherapy. However, little is known about their microbiology and chemistry. In this study, we aim at evaluating the antimicrobial activity of Ma'in hot-springs water (MHSW), studying its microbiology, and determining its physicochemical properties including the heavy metals content. Therefore, water samples were collected from Ma'in hot springs and tested for antimicrobial activity using agar diffusion method. Water was then cultivated on nutrient agar to isolate and identify the dominant bacteria by chemical and molecular methods. The identified strains were tested by cross streak method to evaluate their antimicrobial activity against different clinical and standard strains. Finally, water samples were chemically analyzed and the heavy-metals content was assessed. Results revealed that MHSW was not active against any of the clinical isolates. Nevertheless, MHSW was found to be active against five standard bacterial strains, namely, Staphylococcus epidermidis ATCC 12228 (inhibition zone: 20mm), Staphylococcus aureus ATCC 29213 (inhibition zone: 19mm), Micrococcus luteus ATCC 9341 (inhibition zone: 15.3mm), and Bacillus cereus ATCC 11778 (inhibition zone: 12.3mm). After cultivation of MHSW, five bacterial isolates were obtained and identified based on 16S rRNA gene analysis as new strains of Anoxybacillus flavithermus (identity percentage ranges between 96-99%). Physicochemical analysis revealed that the in situ temperature was 59°C, pH was 7.8, salinity was 1.6ppt, and dissolved oxygen was 3.8mgl-1. In respect to heavy-metals content in MHSW, the following metals were present in the order: Cr (0.571ppm)>Mn(0.169ppm)>Fe (0.124ppm)>Zn (0.095)>Cu(0.070ppm)>Ni(0.058ppm)>Cd (0.023ppm)>Pb (0ppm). Cd, Cr, Ni and Mn were found to be higher than permissible levels set by international organizations and countries. This study highlights new chemical and microbiological data about Ma'in hot springs.

Solid waste characterization, quantification and management practices in developing countries. a case study: Nablus district - Palestine.

Solid waste management (SWM) is one of the most challenging issues faced by developing countries that suffer from serious pollution problems caused by the generation of large waste quantities. This paper presents the case study of SWM in the Nablus district - Palestine. Surveys for household residents' and SWM program operators, field investigations, on-site waste measurements and characterizations were conducted. Per capita waste generation rates varied between different localities although trends were similar. Overall, the majority of waste was organic (65.1% by weight), suggesting a strong resource recovery potential in terms of animal feed or compost. Recyclable waste (plastic, paper and card) made up 16.7% by weight the waste composition suggesting an incentive to introduce source separation. Household attitudes complemented the waste characterization study, revealing the main problems faced. SWM operators quoted on the current status, highlighting problems with disposing in unsanitary landfills, ineffective solid waste fees system, increasing solid waste quantities and lacking equipment and experienced personnel. To enhance sustainable SWM, public awareness, funding, expertise, equipment and facilities and other provisions currently lacking or inappropriate must be provided.

Age-related structural modulation of T lymphocyte-associated CD45 isoforms.

A monoclonal antibody, specific to all conventional CD45 isoforms, was employed in two-dimensional (2D) sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting to investigate possible age-related differential expression of these isoforms among immature and mature thymocytes as well as CD4+ and CD8+ T cell subpopulations in the periphery of newly-born, young and aged BALB/c mice. In young mice, and to a lesser degree in newly-born mice, intra-thymic maturation seemed to be paralleled by the capacity of thymocytes to synthesize distinct CD45 isoforms constituted by extensively heterogeneous acidic charge entities. Thymocyte maturation in aged mice, on the other hand, was characterized by minimal heterogeneity, as the observed pattern was essentially similar to the immature population in 2D blots. As inferred from comparisons of 2D blots of sialylated and desialylated forms of the CD45 complex, age-related differences in isoforms expressed by the CD4+ and the CD8+ T cell subpopulations in the periphery resided mainly in the degree of sialylation of the constituent isoforms. Given the potential of the differential sialylation state of CD45 in altering the recognition properties of lymphocytes, regulation of CD45 sialylation with age may add another level of complexity to the lymphocyte surface phenotype, which in turn may be implicated in cell-cell interaction mechanisms during lymphocyte maturation and senescence.

Identification and localization of a schistosome-associated fucosyllactose determinant expressed by Fasciola hepatica.

A Biomphalaria alexandrina-derived lectin (BaSII), of proven specificity to a Schistosoma mansoni-associated fucosyllactose [(Fuc alpha1-2) Gal beta1-4 Glc] determinant, was employed to investigate the putative antigenic cross-reactivity between Schistosoma mansoni and Fasciola hepatica in terms of this structurally-defined oligosaccharide sequence. BaSII affinity column chromatography of extracts of adult worms metabolically radiolabelled with 35S-methionine and analysis by two-dimensional gels established the expression of the fucosyllactose determinant in multiple copies among heterogeneous, acidic glycoproteins synthesized by adult Fasciola hepatica. Direct fluorescence microscopy revealed that determinant-bearing glycoproteins were localized to the external glycocalyx and perikarya of the tegument as well as the epithelial lining of the intestinal caeca and vitelline ducts and glands. Determinant expression was also evident in embryonated cells of eggs and miracidia as well as the intermediate cellular wall of encysted metacercariae, suggesting its conservation during the course of development of the parasite. Based on the structural relatedness of the cross-reactive fucosyllactose determinant to the antigenic mammalian blood group H trisaccharide, our observations may have implications in serodiagnosis and immunoprophylaxis of schistosomiasis/fascioliasis.

Effect of dietary fat on toad liver tumor induced by DMBA: ultrastructural studies.

Toads injected with 2 mg 7,12-dimethyl-benza(a)anthracene (DMBA)/toad, 3 times/week for 12 weeks induced liver tumors in 12 out of 50 cases. The electron micrograph of toad liver tumor showed disorganization of the rough endoplasmic reticulum which encircles or partially surrounds the mitochondria. Cristae mitochondrialis are rare in comparison with control. Enhancement of liver tumor incidences (29 out of 50 cases) by DMBA at the same dose level plus 2cc corn oil/toad, 3 times/week for 12 weeks was detected. Electronicrograph of this group showed the same criteria of malignancy as in the first group. No tumor incidences were detected in toads fed corn oil only. The electronmicrograph of liver cells showed a high increase in glycogen and lipid droplets.

Small area variation analysis. Methods for comparing several diagnosis-related groups.

In small-area variation analysis, the variation of health care utilization rates, e.g., admission rates, among small areas is calculated. Frequently, the variation of one diagnosis, diagnosis-related group (DRG), or procedure is compared with the variation of another. Unfortunately, the methods generally used to make these comparisons are not consistent. They differ on whether they 1) adjust for the prevalence of the DRGs, 2) distinguish between variation among areas and variation within areas, 3) weight all areas equally, and 4) adjust for multiple admissions per person. None has an associated confidence interval. These discrepancies occur in part because there is no statistical model of small area variation. Without such a model, it is not known how to measure variation, and thus, it is not known how to compare different DRGs. Here, the authors use data on 473 DRGs from 28 counties in Washington state to study the nature of variability. The variation was higher for the more prevalent DRGs, suggesting that adjusting for prevalence may be reasonable. The true coefficient of variation appears to be a "natural" measure of variation, but the usual small area variation statistics do not provide good estimates of the true coefficient of variation. A new estimate is proposed that can be used to compare and test the variability of several DRGs.