Microbial communities in the Dead Sea and their potential biotechnological applications.

The Dead Sea is unique compared to other extreme halophilic habitats. Its salinity exceeds 34%, and it is getting saltier. The Dead Sea environment is characterized by a dominance of divalent cations, with magnesium chloride (MgCl2) levels approaching the predicted 2.3 M upper limit for life, an acidic pH of 6.0, and high levels of absorbed ultraviolet radiation. Consequently, only organisms adapted to such a polyextreme environment can survive in the surface, sinkholes, sediments, muds, and underwater springs of the Dead Sea. Metagenomic sequence analysis and amino acid profiling indicated that the Dead Sea is predominantly composed of halophiles that have various adaptation mechanisms and produce metabolites that can be utilized for biotechnological purposes. A variety of products have been obtained from halophilic microorganisms isolated from the Dead Sea, such as antimicrobials, bioplastics, biofuels, extremozymes, retinal proteins, colored pigments, exopolysaccharides, and compatible solutes. These resources find applications in agriculture, food, biofuel production, industry, and bioremediation for the detoxification of wastewater and soil. Utilizing halophiles as a bioprocessing platform offers advantages such as reduced energy consumption, decreased freshwater demand, minimized capital investment, and continuous production.

Pomegranate peel: Bioactivities as antimicrobial and cytotoxic agents.

This is a comparative study to evaluate the effectiveness of six pomegranate peel extracts (PPEs) as antibacterial and antiproliferative agents. The Six PPEs were prepared using four solvent systems and each filtrate was concentrated to a gummy material to be used in the evaluation. The well-diffusion method was used to evaluate their antimicrobial activity against bacteria typically associated with food spoilage: Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Listeria monocytogenes, Staphylococcus epidermidis, Staphylococcus aureus, and three Bacillus species. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTT) was used to evaluate the cytotoxicity against colorectal carcinoma cells (HCT116), prostate adenocarcinoma (PC3), ovarian cancer cells (SKOV-3), and fibroblasts (MRC-5). The antioxidant evaluation was done using the 2,2-diphenyl-1-picrylhydrazyl-hydrate (DPPH) assay. The pH of the water-containing extracts was acidic and almost the same over 6 weeks. The six PPEs inhibited the bacterial growth in a comparable level to standard antibiotics. The effectiveness of each extract was dependent on the bacterial strain, and the Listeria showed a remarkable inhibition when exposed to the aqueous extract prepared at room temperature (RT). The aqueous (RT) and methanol PPEs had a significant antioxidant scavenging capability and a remarkable cytotoxic activity against the PC3 with half maximal inhibitory concentration (IC50) of 0.1 µg/mL. The boiled aqueous extract exhibited antiproliferative activity against HCT116 with an IC50 of 21.45 µg/mL. The effect on SKOV-3 and fibroblasts was insignificant. With the exception of butanol, the antioxidant screening shows an inverse correlation between the polarity of the extraction solvent and the IC50 exhibited by the PPEs. The variation in the effectiveness of PPEs is suggested to be due to variable soluble bioactive compounds that may interact differently with different cells, though water-containing extracts are promising antibacterial agents. The findings clearly show that pomegranate peel possessed the potential to be an eco-friendly novel source for natural compounds that can be implemented in the food industry as a natural antimicrobial and natural food additive to prevent foodborne illnesses.

Genetic Signature and Serocompatibility Evidence for Drug Resistant Campylobacter jejuni.

Campylobacteriosis, a foodborne illness, is one of the world's leading causes of gastrointestinal illness. This study investigates the link between human campylobacteriosis and the consumption of potentially contaminated food with Campylobacter jejuni. Three hundred sixty samples were collected from humans, chicken cloaca, raw chicken meat, unpasteurized milk, and vegetables. The chickens were obtained from licensed and non-licensed slaughterhouses, and only the necks and wings were studied. Samples were enriched under microaerobic conditions then cultured on the modified charcoal cefoperazone deoxycholate agar. Bacteria was identified by staining, biochemical testing, and molecular identification by the polymerase chain reaction for the virulence genes; hipO, asp, dnaJ, cadF, cdtA, cdtB, and cdtC. The genomic homogeneity of C. jejuni between human and chicken isolates was assessed by the serological Penner test and the pulse field gel electrophoresis (PFGE). Campylobacter was not detected in the vegetables and pasteurized milk, though, only twenty isolates from chickens and clinical samples were presumed to be Campylobacter based on their morphology. The biochemical tests confirmed that five isolates were C. coli, and fifteen isolates were C. jejuni including two isolates from humans, and the remaining were from chickens. The colonization of C. jejuni in chickens was significantly lower in necks (6.66%) obtained from licensed slaughterhouses compared to those obtained from non-licensed slaughterhouses (33.3%). The antimicrobial susceptibility test showed that all identified C. jejuni isolates were resistant to antibiotics, and the majority of isolates (53.5%) showed resistance against six antibiotics, though, all isolates were resistant to ciprofloxacin, tetracycline, and aztreonam. The Penner test showed P:21 as the dominant serotype in isolates from humans, necks, and cloaca. The serohomology of C. jejuni from human isolates and chicken necks, wings, and cloaca was 71%, 36%, 78%, respectively. The PFGE analysis of the pattern for DNA fragmentation by the restriction enzyme Smal showed a complete genotypic homology of C. jejuni human isolates and chicken necks compared to partial homology with cloacal isolates. The study brings attention to the need for effective interventions to ensure best practices for safe poultry production for commercial food chain supply to limit infection with foodborne pathogens, including Campylobacter.

Staphylococcusaureus protein A as a means of assessing sperm penetrability in cervical mucus in vitro.

OBJECTIVE: The effectiveness of Staphylococcus protein A (SPA) in improving the penetration ability of sperm and reducing antisperm antibody (ASA) titers in immunologically infertile males was evaluated. METHODS: Seminal fluid samples were obtained from 15 infertile men, and ASA titers were assessed with the latex agglutination test. Identification of immunoglobulin (Ig) classes and characterization of the antigens involved in the immune response were performed using indirect immunofluorescence. Local ASAs typically present as a mixture of IgG and IgA classes. The capillary tube penetration method was used to assess the capability of spermatozoa to penetrate the cervical mucus (CM). RESULTS: ASAs associated with the neck region of sperm showed a significantly lower migration distance in the CM of infertile females than ASAs associated with the head or tail segments. ASA-positive seminal fluid exhibited significant increases in the mean migration distance (2.6 ± 1.4 cm vs. 1.54 ± 1.1 cm, respectively; p< 0.001) and sperm concentration (174 ± 121.0 × 10³/mL vs. 101 ± 93.7 × 10³/mL, respectively; p= 0.033) after treatment with SPA compared to pre-treated samples. A significant reduction (p< 0.01) in the recorded ASA titer was detected. CONCLUSION: These results indicate that SPA can be used as a sorting regimen for insemination programs. However, further studies are warranted to assess its influence on pregnancy rate.

Isolation and Characterization of Thermophilic Bacteria from Jordanian Hot Springs: Bacillus licheniformis and Thermomonas hydrothermalis Isolates as Potential Producers of Thermostable Enzymes.

The aim of this study was the isolation and characterization of thermophilic bacteria from hot springs in Jordan. Ten isolates were characterized by morphological, microscopic, biochemical, molecular, and physiological characteristics. Sequencing of the 16S rDNA of the isolates followed by BLAST search revealed that nine strains could be identified as Bacillus licheniformis and one isolate as Thermomonas hydrothermalis. This is the first report on the isolation of Thermomonas species from Jordanian hot springs. The isolates showed an ability to produce some thermostable enzymes such as amylase, protease, cellulose, gelatins, and lecithin. Moreover, the UPGMA dendrogram of the enzymatic characteristics of the ten isolates was constructed; results indicated a high phenotypic diversity, which encourages future studies to explore further industrial and environmental applications.

Evaluation of Serum Testosterone, Progesterone, Seminal Antisperm Antibody, and Fructose Levels among Jordanian Males with a History of Infertility.

Due to the biochemical complexity of seminal fluid, we attempt to study the possible correlation between fructose, which is secreted under the effect of androgen hormone, and autoimmunity, which might play a role in varicocele associated infertility, in reducing sperm motility. Seminal fructose, antisperm antibodies (ASAs) and blood steroids hormones (testosterone and progesterone) levels were measured in 66 infertile males with varicocele and 84 without varicocele referred for fertility treatment. Seminal analysis was performed with biochemical measurements of seminal fructose and mixed agglutination reaction (MAR) for ASA. Serum levels of progesterone and testosterone were estimated using a competitive chemoluminescent enzyme immunoassay. The mean values for serum testosterone were 380.74 ± 24.331, 365.9 ± 16.55, and 367.5 ± 21.8 ng/dl, progesterone 0.325 ± 0.243, 0.341 ± 0.022, and 0.357 ± 0.0306 ng/ml, and seminal plasma fructose 359.6 ± 26.75, 315.6 ± 13.08, and 332.08 ± 24.38 mg/dl in males with varicocele, without varicocele, and fertile males, respectively. A significant high level of testosterone was observed within varicocele group (P = .001). This result showed that testosterone may play a role as an infertility determinant in subjects with varicocele. ASA was detected in 18 (26.47%) of cases with varicocele, 20 (38.46%) without varicocele, and in 16 (32.0%) fertile men. Cases with ASAs associated with low sperm motility morphology. An inverse correlation between sperm-bound antibodies and viscosity has been shown (P = .017). ASA showed some significant inverse relations with ages, durations of infertility, and viscosity (P < .05). In addition, a significant correlation was observed between ASA positive seminal plasma and testosterone concentration among infertile cases (with or without varicocele) and fertile (P < .05). Our results suggest a relationship between testicular steroid hormone levels with autoimmunity and sperm antibodies which influence the motility of ejaculated spermatozoa among Jordanian infertile males.

Diagnostic value of various urine tests in the Jordanian population with urinary tract infection.

We compared the performance of leukocyte esterase and nitrite reductase dipstick tests with microscopic examination and uroculture in cases with clinically suspected urinary tract infection (UTI). We studied urine specimens from 504 Jordanian patients which were obtained by the mid-stream clean catch method and analyzed for bacteria. All samples were subjected to culture. Results of urine dipstick tests and pyuria (white blood cells (WBC)/high power field) were compared with urine culture for each sample. Significant bacteriuria was found in 117 cases (23.2%) with positivity of 59% and 68.5% for the presence of nitrite reductase and leukocyte esterase, respectively. Echerichia coli was the most common organism isolated. The dipstick leukocyte esterase and nitrite testing had a sensitivity of 68.5% and 59% for detecting bacteriuria in UTI cases and specificity of 73.5% and 78%, respectively. The positive predictive value of the tests was 44% and 60%, and the negative predictive value 88.5% and 86.2%, respectively. Microscopic WBC showed 86.5% specificity but low sensitivity. Urine dipstick results and pyuria significantly correlated with the results of urine culture but demonstrated more false-positive results, which ranged from 13.4-26.6%. The probability of growing a urinary pathogen correlated with urinary WBC counts and allowed prediction of the presence or absence of bacteriuria by counting urinary leukocytes. A combination of pyuria and urine dipstick testing appears to be a very useful marker for the diagnosis of UTI. Urine culture can be omitted if both tests are negative.