Open Access Case Series

Evaluation of Different Screening Methods for Biosurfactant Producers Isolated from Contaminated Egyptian Samples Grown on Industrial Olive Oil Processing Waste

N. M. Sidkey, H. F. Mohamed, H. I. Elkhouly

Microbiology Research Journal International, Page 1-19
DOI: 10.9734/BMRJ/2016/28437

The present study aimed to screen biosurfactant producing microorganisms isolated from different Egyptian samples viz. soil sample contaminated with oil from fuel station, soil sample contaminated with kerosene from classic bread oven, samples from wall of drainage tube of kitchen and bathroom, also waste sample from gas cooktops of kitchen stove. All isolation samples were streaked on MSM medium supplemented with 1% olive oil processing waste as a sole carbon source to recover bacterial isolates with biosurfactant activity. Different screening methods e.g. Oil spreading assay, Emulsification index E24, Drop collapse test, Blue agar plate method (Cetyl trimethyl ammonium bromide-CTAB), Blood agar haemolysis, Reduction in SFT and Phenol sulfuric acid method were used to screen microbial biosurfactant producers. Fifty five bacterial isolates were obtained, consecutive screening was carried out between isolates to select the most promising biosurfactant producer. The selected isolate produced potential biosurfactant that belongs to glycolipid and identified by Biochemical and 16S rRNA analysis and was found belongs to Pseudomonas aeruginosa PAO1.

Conclusion: A combination of different methods is required for a successful screening, but it is recommend using both of drop collapse and CTAB tests, suggesting that strains highly active in one method were active in the other method. In addition, surface tension measurement and phenol sulfuric acid reaction is a must in case of the biosurfactant is of glycolipid type (rhamnolipid) to confirm the presence of anionic biosurfactant. In the present investigation using of efficient biosurfactant producer (P. aeruginosa PAO1) which prefer limited oxygen requirements (microaerobic) growing on low cost substrate (olive oil processing waste) is a privilege in the production cost.

Open Access Short Research Article

Anti-thyroid Antibodies in Patients with HCV Genotype 3a: A Pilot Study

Sana Temuri, Nadeem Afzal, Mohammad Kashif, Ahmed Nadeem, Faheem Shahzad, Waqas Latif, Afia Abbas, Tafazzul H. Mahmud

Microbiology Research Journal International, Page 1-5
DOI: 10.9734/BMRJ/2016/28690

Background: Each year, 3 to 4 million people are infected with hepatitis C virus (HCV) and it is the major cause of liver disease worldwide. The patient with acute HCV is often asymptomatic but can present with fatigue and jaundice. About 80% of HCV infected individual’s progress to chronic state. There is an increased prevalence of HCV infection with autoimmune diseases and the most common is chronic thyroiditis, which is an inflammatory disease that leads to hypothyroidism. These individuals have an increased level of antithyroid peroxidase antibody (anti TPO-Ab).

Objectives: To determine the prevalence of antithyroid antibody (ATA) in patients of HCV genotype 3a.

Materials and Methods: Fifty HCV infected patients with genotype 3a were enrolled in this study that included 33 males and 17 females. After written informed consent, 3 ml blood of all the patients was obtained and ATAs were detected by indirect immunofluorescence technique. These patients were divided into 3 groups, i.e. untreated 26 (52%), mid treated 17 (34%) and 7 (14%) patients with hepatocellular carcinoma (HCC).

Results: Among the patients 17(51.5%) males and 7 (41.2%) females had ATA. Regarding different groups, 19 (73.1%) of untreated, 5 (29.4%) of mid treated and none of the patient in HCC group had ATA.

Conclusion: ATA was detected in a high percentage of patients with HCV genotype 3a. These antibodies were significantly higher in untreated patients as compared to mid treated and HCC patients. Further, more males had these antibodies as compared to females.

Open Access Original Research Article

Identification of Ascitic Fluid Bacterial Pathogens in Spontaneous Bacterial Peritonitis in Nile Delta and Its Impact on Clinical Outcome of these Patients

Haidy S. Khalil, Walaa Elkhalawany, Mohamed Elhendawy, Rehab Badawi, Marwa Ahmed Abdelwahab, Sherief Abd- Elsalam

Microbiology Research Journal International, Page 1-6
DOI: 10.9734/BMRJ/2016/29869

Aims: This study aimed to identify ascetic fluid bacterial pathogens and their antibiotic resistance profile in Spontaneous Bacterial Peritonitis (SBP) patients in Nile delta and its impact on the clinical outcome of these patients.

Study Design:  Retrospective observational study.

Place and Duration of Study: Patients enrolled in this study were admitted to Tropical Medicine and Infectious Diseases Department, Faculty of Medicine, Tanta University, Egypt. Further laboratory work was carried out at Microbiology and Immunology Department, Faculty of Medicine, Tanta University, Egypt, from July 2015 to June 2016.

Methodology: 247 patients with liver cirrhosis and ascites who met the clinical criteria for suspicion of SBP including:  fever, encephalopathy, refractory ascites and abdominal pain were enrolled in the study. Patients were subjected to thorough history and clinical examination. Ascetic fluid sampling was done for every patient and ascetic fluid analysis was done including cell counts and differential counts. Also, ascitic fluid culture, microbiological testing and antimicrobial sensitivity tests were done.  

Results: Out of 247 patients enrolled in this study with liver cirrhosis, ascites and clinical suspicion of SBP, 138 patients were excluded. These excluded patients included: 91 patients had ascetic fluid neutrophils below 250 cells/mm3, 4 patients were cases of secondary peritonitis with polymicrobial culture and 43 patients were found to started empirical antibiotics within 5 days of admission. Out of 109 patients who had SBP, 28 only were culture positive. Among culture positive SBP, 16 (57.1%) were Gram positive and 12 (42.9%) were Gram negative. The most common organism isolated was Gram positive Enterococci followed by E. coli and Staph aureus.

Conclusion: While Gram negative bacteria were the main infectious agents causing SBP a few decades ago, and are still reported to be so in the most recent recommendations and reviews, Gram positive bacteria are now predominant and there is a rising prevalence of bacteria with reduced susceptibility to cephalosporins and fluoroquinolones as regarding this study only and not including previous data or speculations. Current international guidelines recommend the use of a third-generation cephalosporin for empirical treatment of SBP which raise the questions about these guidelines and if they are still valid.

Open Access Original Research Article

Antibacterial Activities of Essential Oils from Three Medicinal Plants in Combination with EDTA against Methicillin-resistant Staphylococcus aureus

Juan Rojas Armas, Julio Ruiz Quiroz, Robert Almonacid Roman, Jose Ortiz Sanchez, Miriam Palomino Pacheco, Luz Huaroto Valdivia, Emilio Collahua Rivera, Roberto Chavez Asmat, Andrea Anampa- Guzmán

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/BMRJ/2016/29666

Aims: To determine the antibacterial activity of essential oils (EOs) from Thymus vulgaris L, Origanum vulgare L and Minthostachys mollis (Benth.) Griseb in combination with ethylenediaminetetraacetic acid (EDTA) on methicillin-resistant Staphylococcus aureus (MRSA) from clinical isolates.

Study Design: Collection of plant material, extraction, phytochemical analysis and evaluation of antimicrobial activity of the essential oils.

Place and Duration of Study: Clinical Research Institute, Faculty of Medicine, Universidad Nacional Mayor de San Marcos, Department of Microbiology, Faculty of Pharmacy and Biochemistry, and Department of Microbiology, Hospital Dos de Mayo, Lima, Peru, between February 2014 and February 2015.

Methodology: The EOs’ chemical composition was determined by gas chromatography coupled with mass spectroscopy (GC-MS). Clinical isolates of MRSA were obtained from the “Dos de Mayo” National Hospital in Lima, Peru. The inhibitory activity on MRSA was determined by disk diffusion method and Minimum inhibitory concentration (MIC) by the microdilution colorimetric method in 96-well plates. All statistical analysis was performed using SPSS V 21.0 software.

Results: The main components of Thymus vulgaris EO were thymol (46.47%), γ-terpinene (20.27%) and p-cymene (15.80%); from Origanum vulgare EO were γ-terpinene (21.17%), (-)-4-terpineol (12.61%) and cis-β-terpineol (12.18%); and from Minthostachys mollis EO were pulegone (33.48%) and menthone (26.68%). The Thymus vulgaris EO presented inhibition halo diameters of 32.23 ± 1.70 mm, and Thymus vulgaris + EDTA had inhibition halo diameters of 32.47 ± 2.06 mm, both significantly higher compared to 30 mcg of cefoxitin, which had inhibition halo diameters of 17.60 ± 0.68 mm (p < .01). The MIC of T. vulgaris against MRSA was 0.57 µg/mL. Origanum vulgare and Minthostachys mollis EOs were resistant to MRSA.

Conclusion: This study showed that the Thymus vulgaris essential oil had antibacterial activities against MRSA and indicates their potential application in the control of this pathogen commonly known for their resistant activities to most conventional antibiotics.

Open Access Original Research Article

Molecular Characterization of a Sweet Potato Leaf Curl Virus Isolate from Egypt and Its Phylogenetic Relationship with Other Members of the Begomovirus

Aly M. Abdel- Salam, Malik - Mujaddad-ur Rehman, Salama M. El- Saghir, Manal A. El- Shazly

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/BMRJ/2016/29263

The present study was undertaken to isolate and molecularly and phylogenetically characterize a Sweet potato leaf curl virus isolate infecting sweet potato plants in Giza governorate in Egypt. The virus was isolated from infected plants showing leaf curl symptoms using non-viruliferous Bemisia tabaci insects. Total extracted DNA was amplified with polymerase chain reaction (PCR) using the degenerate AV/AC core primers and produced 580 bp of the DNA-A core coat protein in agarose gel. PCR conditions of 6 mM MgCl2, combinatorial enhancer solution, and a 53°C/40s annealing temperature facilitated coat protein amplification. Recovered virus amplicon was ligated into pGEM T-Easy vector and cloned into Eschericia coli, strain DH5α. Purified plasmid DNA was submitted to GenBank and given an accession number FJ455517. Sequence comparisons with 13 sweepoviruses indicated that the SPLCV-Giza isolate has the highest nucleotide sequence identity (97%) with the SPLCV-US (KC253238). According to the current taxonomic criteria for Begomovirus classification, the Giza isolate of SPLCV in Egypt would be considered as a variant of the SPLCV-US. Phylogenic study using amino acid substitution showed the clustering of the 13 studied sweepoviruses in one major clade and one minor clade.  The SPLCV-Giza isolate clustered with SPLCV-US in a monophyletic branch within the large clade circumventing nine sweepoviruses, viz. SPLCV isolates from: Korea, Brazil, Japan, USA, China, India, and Ipomoea yellow vein virus (IYVV) from Italy and Spain. The second minor clade involved the clustering of SPLCV isolates from Uganda and Spain in a monophyletic branch apart from the rest of the studied sweepoviruses.  Phylogenic analysis of SPLCV-Egypt with five other mono and bipartite begomomoviruses showed its distinctive clustering nature from these viruses. In conclusion, the present study confirms the presence of an isolate SPLCV in Egypt as a variant of the SPLCV-US. The study also illustrates the high worldwide diversity of SPLCV isolates and signifies the economic importance of this newly introduced virus into Egypt. 

Open Access Original Research Article

Probiotic Properties Profiling of Isolated Lactic Acid Bacteria from the Intestine of Channa punctata

Rifat-Al- Naser, Md. Abdullah-Al- Mamun, Md. Saddam Hossain, Forhad Karim Saikot, Saifullah ., Md. Arifuzzaman

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/mrji/2016/v17i45181

Aims: This study was conducted for isolation, identification and probiotic profiling of Lactobacillus spp. from the intestine of indigenous and non-indigenous Channa punctata species.

Study Design: Lactobacillus spp. from the intestine of Channa punctata fish was isolated and cultured as well as screened for probiotic properties and profiling.

Place and Duration of Study: The research work was carried out in Biotechnology Laboratory, Department of Genetic Engineering and Biotechnology, Jessore University of Science and Technology, Jessore-7408, Bangladesh, from July to October, 2015.

Methodology: Lactic Acid Bacteria (Lactobacillus spp.) were isolated from the intestine of dissected Channa punctata by using MRS agar medium following spread plate method. Isolated bacteria were subjected to gram staining, microscopic observation and biochemical tests example catalase for identification of Lactobacillus spp. Thereafter, bile salt tolerance test, phenol tolerance test and antibiotic sensitivity test were done for probiotic profiling.

Results: Two Lactic Acid Bacteria were isolated from indigenous and non-indigenous Channa punctata species where total bacterial cell was counted as 2.1×1010 and 1.9×10cfu/gm. Both the fish intestines contain bacteria having gram positive, non-sporulating, catalase negative rods which were confirmed to be Lactobacillus. They were able to survive in bile salt and phenol at 0.1, 0.2, and 0.3% concentration.  The Lactobacillus isolated from indigenous Channa punctata was resistant to Azithromycin, Cefuroxime, Ciprofloxacin, Tetracycline, Ampicillin, Erythromycin, Vancomycin, Chloramphenicol and Co-trimoxazole while Lactobacillus from non-indigenous C. punctata showed almost same resistant except Cefotaxime and Gatifloxacin. They are claimed as probiotics.

Conclusion: Thus, the study claimed that the isolated, counted and identified Lactobacillus spp. are belong to probiotic that can exert beneficial health action for human.