Comparative molecular studies of halophilic bacteria from saline water and soil in the Saudi environment

Halophilic bacteria are a microorganism that grows optimally in the presence of the very high concentration of sodium chloride. Halophiles are vital sources of various enzymes including hydrolases, which are very stable and catalytically highly efficient at high salt concentration and other extreme conditions such as high temperature, pH and presence of organic solvents.  Several hydrolases such as amylases, proteases, and lipases have been obtained from halophilic bacteria and are commonly used for various industrial applications. We initiated a screening project to isolate and characterize the halophilic bacteria from the Red Sea, which is one of the saltiest bodies of water in the world. Water and soil samples, collected from the Red Sea coast, Jeddah, Saudi Arabia, were screened for isolation of halophilic bacteria. Ten bacterial isolates were obtained, which were characterized by biochemical tests and 16S rRNA gene sequencing. Hydrolase producing bacteria among the isolates were screened by plate assay on starch and gelatin agar plates for amylase and protease, respectively.  Two bacterial isolates i.e Bacillus haynesii and Enterobacter cloacae subsp. were found to possess significant amylase and protease activity. Further characterization of both the strains is in progress

Vaccination against Corynebacterium pseudotuberculosis infections controlling caseous lymphadenitis (CLA) and oedematousskin disease

Corynebacterium pseudotuberculosis (C. pseudotuberculosis) is a causative organism of caseous lymphadenitis (CLA) in sheep and acute disease in buffaloes known as oedematous skin disease (OSD). Human affected with the disease show liver abscess and abscess in the internal lymph nodes. The vaccination against CLA up till now occurs by using formalin inactivated whole cells of biovar 1 (sheep strain). Combined vaccine composed of formalin inactivated whole cells of sheep strain and recombinant phospholipase D (rPLD) and another vaccine composed of formalin inactivated whole cells (buffalo origin) and rPLD were prepared in Biotechnology center for services and Researches laboratory at Cairo university and applied for protection against CLA. Both vaccines induced complete protection (100%) against challenge with virulent biovar 1 or biovar 2. Also vaccination against OSD was performed by two types of vaccines. Vaccine-1 was composed of formalin inactivated whole cell biovar 1 combined with rPLD and the second vaccine was composed of formalin inactivated whole cells of biovar 2 combined with rPLD. No lesions developed in vaccinated and non vaccinated buffaloes challenged with C. pseudotuberculosis biovar revealing that biovar 1 C. pseudotuberculosis is not infective for buffaloes. Buffaloes vaccinated with the second vaccine and control non vaccinated animals challenged with biovar 2 (buffalo origin) resulted in development of OSD in all animals. This indicates that OSD results due to production of toxin (s) other than PLD. Discovering this toxin (s) is of value in formulation of a future vaccine against OSD. Keywords: Corynebacterium pseudotuberculosis, Caseous lymphadenitis, Oedematous skin disease, Vaccination, Recombinant phospholipase

Therapeutic role of Ricinus communis L. and its bioactive compounds in disease prevention and treatment

Ricinus communis L. (R. communis), commonly known as castor oil plant, is used as a traditional natural remedy or folkloric herb for the control and treatment of a wide range of diseases around the globe. Various studies have revealed the presence of diverse phytochemicals such as alkaloids, flavonoids, terpenes, saponins, phenolic compounds such as kaempferol, gallic acid, ricin, rutin, lupeol, ricinoleic acid, pinene, thujone and gentisic acid. These phytochemicals have been responsible for pharmacological and therapeutic effects, including anticancer, antimicrobial, insecticidal, antioxidant, anti-diabetic, antinociceptive, anti-inflammatory, bone regenerative, analgesic, and anticonvulsant activity. R. communis harbours phytochemicals which have been shown to target peroxisome proliferator activated receptor (PPAR), nuclear factor NF- κ -B, cytochrome p450, P38 mitogen-activated protein kinases kinase (p38 MAPK), tumor protein P53, B-cell lymphoma-extra-large (Bcl-xL) and vascular endothelial growth factor receptor-2 (VEGFR-2). Considering its wide variety of phytochemicals, its pharmacological activity and the subsequent clinical trials, R. communis could be a good candidate for discovering novel complementary drugs. Further experimental and advanced clinical studies are required to explore the pharmaceutical, beneficial therapeutic and safety prospects of R. communis with its phytochemicals as a herbal and complementary medicine for combating various diseases and disorders

An alternative approach for evaluating the phenotypic virulence factors of pathogenic Escherichia coli

Escherichia coli is a recognized zoonotic food-borne pathogen; however, the use of polymerase chain reaction (PCR) in the underdeveloped countries to differentiate pathogenic from non-pathogenic E. coli is a problematic issue. Our grail was to assess the phenotypic virulence markers motility, hemolysin, congo red agar, embryo lethality assay and serum resistance for pathogenic E. coli (PEC) correlated to PCR tests which is currently used world-wide to evaluate the PEC. The 448 strains of Escherichia coli that were isolated from different sources, were characterized for phenotypic virulence factors such as motility, hemolysin, Congo red binding, Embryo Lethality assay (ELA) and serum resistance, as well as antibiotic susceptibility using disc diffusion method to 23 antibiotics. Results exhibited 100% motility and Congo red binding, 97.1% for hemolysin production and 90.2% in the ELA. As a result, we were able to hypothetically conclude that the aforementioned virulence markers are plain, straightforward, economical, rapid, more dynamic, uncomplicated methodology, duplicatable and cost next to nothing when compared to the molecular PCR. Their implementation in a diagnostic microbiology laboratory for vetting is a rewarding task in the underdeveloped countries. It augments endeavors to minimize the use of PCR in our investigations especially during epidemiological and outbreak investigations of PEC

Molecular characterization of the capsular antigens of Pasteurella multocida isolates using multiplex PCR

The use of molecular techniques for detection and characterization of the Pasteurella multocida is very important for rapid and specific detection and characterization of the organism. During the period from 15th February, 2014 to 15th April, 2015, 425 nasopharyngeal swabs and 175 lung and spleen samples were collected and examined by conventional methods, 80 strains (18.82%) of P. multocida were isolated from the calves, sheep and goat with respiratory manifestation. Meanwhile, 77 strains (44%) were isolated from emergency slaughtered animals. All the recovered strains were positive for specific PCR for detection of P. multocida strains previously identified as P. multocida by standard microbiological techniques. Multiplex PCR for molecular typing of the capsular antigens of the recovered P. multocida revealed positive amplification of 1044 bp fragments specific to the capsular antigen type A with 105 strains (66.88%), and amplification 511 bp fragments of the capsular antigen type E with 52 strain (33.12%) and absence of B, D and F antigens. Multiplex PCR for molecular typing of the capsular antigens of P. multocida can be used as a simple, sensitive, rapid, reliable technique instead of the serological techniques for identification of the capsular antigens of P. multocid

Zoonotic risk and public health hazards of companion animals in the transmission of Helicobacter species

Objective: Helicobacteriosis is worldwide infection caused by Helicobacter species that affects both humans and animals. The current work correlated the zoonotic and public health repertoire of Helicobacter species in companion animals (dogs and cats). Methods: Samples were collected from apparently healthy dogs (70), cats (65), and 70 human patients who had been in contact with these animals in the Cairo and Giza governorates. The samples included serum, feces, and stool samples and biopsies of gastric fundus fragments (~5 mm). All samples were examined by culture, biochemical analysis, serology, and molecular identification. Results: Helicobacter species were detected at a rate of 43.4% by PCR. H. heilmannii was more predominant, with a rate of 16%, whereas H. pylori was detected at 6%. H. pylori and H. heilmannii were isolated from both human and companion samples, whereas all samples were negative for H. felis. Conclusion: Dogs and cats were reservoirs and played a major source in human helicobacters infection

Recent approaches for control of E. coli and respiratory complex in Middle East

This study was conducted on 100 one-day-old broiler chicks to evaluate the effect of Poulvac E. coli vaccine in reduction of clinical signs and complications after concurrent infectious bronchitis virus (variant 02) and virulent E. coli O78 challenges. The birds were evaluated for clinical signs, mortality for 7 days post-infection, PM lesion score, average body weight and serological evaluation. Re-isolation and RT-PCR for the challenging infectious bronchitis virus (IBV) variant 02 were conducted thereafter. The results showed that the Poulvac E. coli at one-day old chicks in the presence of co-infection with virulent E. coli and IBV variant 02 provides better body weight gain at 35 days than the other groups. The challenge with IBV variant 02 alone in non-vaccinated birds doesn’t give any mortality; this indicated that the severity of IBV variant 02 increased by the presence of co-infection with Avian Pathogenic E. coli (APEc). The mortality percentage associated with both E. coli and IBV variant 02 infections in the none vaccinated group by Poulvac E. coli was 25% while this percentage was 10% of the vaccinated group. The Poulvac E. coli is not negatively affecting the immune response against different concurrent viral vaccines like Infectious bursal disease (IBD), and moreover, it improves the immune response against some others like Newcastle disease virus (NDV), Avian Influenza (AI) H5 and IBV. Keywords: Collibacillosis, Poulvac E. coli vaccine, IBV variant 0

Isolation of antimicrobial producing Actinobacteria from soil samples

Emergence of multidrug resistant bacteria has made the search for novel bioactive compounds from natural and unexplored habitats a necessity. Actinobacteria have important bioactive substances. The present study investigated antimicrobial activity of Actinobacteria isolated from soil samples of Egypt. One hundred samples were collected from agricultural farming soil of different governorates. Twelve isolates have produced activity against the tested microorganisms (S. aureus, Bacillus cereus, E. coli, K. pneumoniae, P. aeruginosa, S. Typhi, C. albicans, A. niger and A. flavus). By VITEK 2 system version: 07.01 the 12 isolates were identified as Kocuria kristinae, Kocuria rosea, Streptomyces griseus, Streptomyces flaveolus and Actinobacteria. Using ethyl acetate extraction method the isolates culture’s supernatants were tested by diffusion method against indicator microorganisms. These results indicate that Actinobacteria isolated from Egypt farms could be sources of antimicrobial bioactive substances