Open Access Original Research Article

Molecular Identification, Bioactivity Screening and Metabolic Fingerprinting of the Actinomycetes of Chenab River Sediments

Mohsin Tassawar Cheema, Adeela Fatima, Imran Sajid

Microbiology Research Journal International, Page 1-13
DOI: 10.9734/BMRJ/2016/28805

Aims: The mortality, morbidity and health cost have been increased due to multiple antimicrobial drug resistance in pathogens across the world. So it is important to discover new active compounds. This study was designed to screen the Chenab river actinomycetes for antimicrobial activity against various MDR bacterial pathogens and preliminary cytotoxicity.

Study Design: Collection of water and sediments samples, sample enrichment and selective isolation of the actinomycetes, laboratory scale cultivation, solvent extraction, determination of antimicrobial activity and cytotoxicity, chemical profiling and metabolic fingerprinting by TLC and HPLC-UV.

Place and Duration: All the work was done during July 2013 to July 2015 in the Department of Microbiology and Molecular Genetics, University of the Punjab Lahore, Pakistan.

Methodology: The water and sediments samples of Chenab river were collected from the Sialkot region (Punjab, Pakistan) and were processed for the selective isolation of actinomycetes. The isolated strains were identified by morphological, biochemical, physiological characterization and by 16S rRNA gene sequencing. The selected strains were screened for antimicrobial activity and cytotoxicity using agar diffusion assays and by microwell cytotoxicity assay against Artemia salina larvae. For metabolic fingerprinting the methanolic crude extracts obtained from the selected strains were analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC-UV).

Results: A total of 30 actinomycetes strains were isolated and were identified as different species of genus Streptomyces. Among all the selected strains, the isolates M72, M1, M71, W38, W108 and M93 were found to exhibit potent antimicrobial activity against the test strains including Klebsiella sp., methiciliin resistant Staphylococcus aureus (MRSA), Acinetobacter sp., E. coli, and B. cereus. Similarly, the isolates M54, W19 and M81 demonstrated significant cytotoxicity (up to 80% larval mortality) against Artemia salina.

Conclusion: The study suggested that actinomycetes flora of Chenab river is rich source of useful natural compounds and should be continuously isolated characterized and investigated for novel antibiotics and other chemotherapeutic agents.

Open Access Original Research Article

Screening, Growth Characterization and Alkaline Phosphatase Potential of Pseudomonas plecoglossicida from Mangrove Soil

S. Sankaralingam, B. Harinathan, S. Palpperuma, D. Kathiresan, S. Rajendran, T. Sivakumar, T. Shankar, G. Prabakaran, N. Sivakumar

Microbiology Research Journal International, Page 1-11
DOI: 10.9734/BMRJ/2016/28488

The study was carried out to isolate phosphate solubilizing bacteria from mangrove soil collected from the Manakudi estuary, Kanyakumari, Tamilnadu, India. Around 20 colonies were obtained in Zobell marine agar plates screened on Pikovskaya’s agar plate for determining their phosphate solubilizing ability. Out of these 20 isolates, one isolate exhibited better phosphate solubilizing ability. Identification of strain was carried out and confirmed by cultural, biochemical and 16S rDNA sequences. The potent isolate was identified as Pseudomonas plecoglossicida. Various factors were tested for pH, temperature, carbon and nitrogen sources, tricalcium phosphate influenced bacterial growth and phosphate activity in static and shaking conditions and thus characterized.

Open Access Original Research Article

Antimicrobial Resistance of Pathogenic Bacteria Isolated from Mastitis Cows in Khartoum State, Sudan

W. M. Yasin, Y. A. Sabiel, A. A. El- Gaddal, M. E. Mansour

Microbiology Research Journal International, Page 1-6
DOI: 10.9734/BMRJ/2016/28838

Aims: The problem of antimicrobial resistance (AMR) is now recognized as a major threat to the health and development in all countries. This study was conducted to determine pathogenic bacteria associated with clinical cases of bovine mastitis and their antimicrobial resistance patterns.

Study Design: This study was carried out In the Department of Bacteriology, Central Veterinary Research Laboratory, Khartoum, Sudan during the period from April 2013 to March 2014.

Methodology: 150 milk samples from clinical cases of bovine mastitis were cultured onto blood agar plates and the isolated organisms were identified by conventional bacteriological methods. One hundred and five isolates were tested against 11 antimicrobial agents commonly used in the dairy farms using the disc diffusion method.

Results: The majority of the isolates were highly sensitive to gentamycin, ciprofloxacin, norfloxacin and kanamycin, highly resistance to penicillin-G and moderately sensitive to novobiocin, tetracycline and cefalexin.

Conclusions: This study revealed that a number of significantly public health concern bacteria were isolated from milk samples and increasingly developed resistance to different groups of antimicrobial agents.

Open Access Original Research Article

Antimalarial Efficacy of Bergenia ciliata (Saxifragaceae) Leaf Extract In vitro against Plasmodium falciparum and In vivo against Plasmodium berghei

Neha Sylvia Walter, Upma Bagai

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/BMRJ/2016/29262

Aim: The traditional medicinal plant Bergenia ciliata was used to evaluate its antiplasmodial activity against Plasmodium falciparum in vitro and preventive and curative activity against Plasmodium berghei in vivo. The safety of the ethanolic leaf extract of Bergenia ciliata (ELEBC) to the liver and kidney functions of the rodent host was also tested.

Place and Duration of the Study: Parasitology Laboratory, Department of Zoology, Panjab University, Chandigarh, India, between October 2014 to November 2015.

Methodology: The in vitro antiplasmodial activity of the ELEBC against both chloroquine-resistant (RKL-9) and sensitive (MRC-2) strains of P. falciparum was assessed by using the WHO method. The cytotoxicity of the extract against human cancer and normal cell lines was tested by MTT assay. The in vivo repository and curative efficacy of the extract against P. berghei were tested using the Peter’s method and modified method of Ryley and Peters respectively. The biochemical assays were performed as per standard methods.

Results: ELEBC exhibited considerable inhibitory activity against both RKL-9 and MRC-2 strains of P. falciparum with IC50 of 6.4 µg/ml and <5 µg/ml respectively. The extract exhibited no toxicity against both cancer and normal cell lines with CC50 >1000 µg/ml and selectivity index (SI) >10. Maximum chemosuppression of 74.45% and 91.96% was observed on day 7 at a concentration of 1000 mg/kg (repository activity) and 250 mg/kg (curative activity), respectively. 83.33% survival of mice was observed in G6 (750 mg/kg) while in all other ELEBC treated groups 50% survival was recorded on day 28 of study in the curative test. Hepatic function (SGOT, SGPT, ALP and bilirubin) and renal function biomarkers (creatinine and urea) in serum were observed to be significantly (P< 0.0005) lower as compared to the infected control (G2).

Conclusions: ELEBC possesses considerable antimalarial activity against both sensitive and resistant strains of P. falciparum. It also exhibits significant efficacy as a preventive and curative remedy against the disease without any side effects on hepatic and renal functions of the rodent hos

Open Access Original Research Article

Evaluation of DNA Based Techniques for the Diagnosis of Human Vaginal Trichomoniasis in North Indian Population

Subash Chandra Sonkar, Sonal Yadav, Nancy Malla, Rakesh Singh Dhanda, Sumeeta Khurana, Rashmi Bagga, Daman Saluja, Manisha Yadav

Microbiology Research Journal International, Page 1-12
DOI: 10.9734/BMRJ/2016/29557

Introduction: Human trichomoniasis due to Trichomonas vaginalis is a curable sexually transmitted infection. It may lead to symptomatic vaginitis or asymptomatic carrier state. The symptoms and signs mimic other pathologies and conventional techniques of diagnosis have its own limitations. Therefore, the present study aimed to evaluate a newly established in-house PCR based assay on pfoB gene and compared it with conventional wet smear examination method and 18S rRNA gene based PCR technique for the diagnosis of T. vaginalis in symptomatic and asymptomatic subjects.

Materials and Methods: Four hundred women in age group 20-57 years attending the Obstetrics and Gynecology out Patients Department (OPD) of Nehru Hospital attached to Post Graduate Institute of Medical education and Research (PGIMER) Chandigarh, India were included in the study. Based on the symptoms and signs, 344 (86%) women were categorized as symptomatic and 56 (14%) as asymptomatic. Vaginal swabs collected from all the women were processed by three techniques including wet smear, 18S rRNA and the pfoB gene based PCR techniques for the detection of T. vaginalis.

Results: The main presenting symptom in majority of symptomatic patients was vaginal discharge. The highest numbers of T. vaginalis positive patients were found in the sexually active age group of 20 to 40 years. The amplifications of pfoB and 18S rRNA gene by PCR revealed significantly higher positive cases (20.7% and 18.6%, respectively) than the wet smear (6.6%) method. The diagnostic efficacy and kappa value estimated by the three techniques were 86.8-100% and 35.5-66%, respectively.

Conclusions: The combined application of any two of the three techniques used in the present study may be useful for the diagnosis of T. vaginalis infection in symptomatic and asymptomatic subjects. This information may help clinicians to make a timely and accurate diagnosis.

Open Access Original Research Article

Anaerobic Digestion of Biodegradable Domestic Wastes by Microorganisms

E. U. Eleanya, C. K. Wachukwu, A. O. Ollor

Microbiology Research Journal International, Page 1-9
DOI: 10.9734/BMRJ/2016/21910

The aim of this study was to demonstrate that biogas can be generated from biodegradable domestic wastes and to determine the bacterial succession involved in the anaerobic decomposition of the wastes. Ten kilogram (10 kg) of biodegradable domestic waste was made into slurry with tap water. The slurry was fed into a batch system biodigester and left at room temperature for 12 weeks. Metagenomic method was used to determine the bacterial and archaeal species involved in the anaerobic digestion. MULTIRAE PGM 50 was used to confirm the presence of the generated biogas from the slurry. Serial dilutions of the slurry was made on alternate days and the appropriate dilutions were inoculated onto nutrient agar plates for bacterial isolation and incubation was at 35°C for 48 hrs. Potato dextrose agar was used for fungal isolation, and incubation was at ambient temperature for three days. Pure isolates of representative communities were maintained on agar slants at 4°C. Triplicate samples from various tubes were cultured and the average count was used. Fungal growth occurred on the PDA plate only on the first day of incubation. The mean total bacteriaial count was highest on the second day (1.3 x10cfu/ml); it decreased with increasing incubation time and became constant from the 23rd day to the end of the experiment (1.0 x101 cfu/ml). The microorganisms involved in the biodegradation were found to be Lactobacillus rapi strain LA1165, Clostridyum tyrobutyricum, Ralstonia pickettii, Methanoculleus marisnigri, Methanosarcina acetivorans C2A, Clostridium acetobutylicum EA 2018, Clostridium tyrobutyricum 5S, Halothermothrix oremii H168, Lactobacillus rapi strain LA1165, Lactobacillus buchneri, Solobacterium moorei W540, B. vulgatus ATCC8482. Rhizopus spp and Aspergillus spp were isolated only on the first two days of incubation. The result from this study proves that, it is possible to generate biogas from domestic wastes and diverse species of microorganisms are involved in anaerobic digestion of biodegradable domestic wastes.