Open Access Original Research Article

Cross Sectional Study on Antibiotic Prescription for Acute Respiratory Tract Infection of Children under Age of 5 at Tertiary General Hospital in Jakarta Indonesia

Maksum Radji, Siti Fauziyah, Oktaviani Tika Wulandria

Microbiology Research Journal International, Page 723-729
DOI: 10.9734/bmrj/2014/7835

Aim: The purpose of this study was to evaluate the use of antibiotics in acute respiratory infections in children in Dr Mintohardjo Navy Hospital, Jakarta.

Place and Duration of Study: Pediatric Clinic of Dr. Mintohardjo Navy Hospital, Jakarta, Indonesia during January to December 2012.

Methodology:  This study is a cross-sectional study consisting of children under the age of 5 years, who suffered from acute respiratory tract infections and hospitalized at pediatric clinic of Dr. Mintohardjo Navy Hospital, Jakarta. The data were collected from patient medical records retrospectively. The assessment of antibiotic prescribing patterns for children younger than 5 years was carried out based on the Indonesian Guideline of antibiotic use in acute respiratory tract infections in children.

Results: A total of 96 patients enrolled in this study consisted of 53.1% males and 46.9% females. The types of acute respiratory tract infections were acute pharyngotonsilitis (95.8%), acute pneumonia (3.1%) and acute laryngitis (1.1%). The most commonly used antibiotics were ceftriaxone (42.5%), cefotaxime (30.0%), gentamicin (6.3%), cefadroxil (5.0%), cefixime (5.0%), sulfamethoxazole-trimethoprim (5.0%), amoxicillin (2.5%), thiamphenicol (2.5%) and chloramphenicol (1.3%).

Conclusion: The compliance rate of pediatricians to follow the Indonesian Guideline on the use of antibiotics for acute respiratory tract infections was very low. It is necessary to increase compliance with the Indonesian Guideline to improve the control program of acute respiratory infections, and to prevent the emergence of antibiotic resistance.

Open Access Original Research Article

Comparison of Use of Agglutination of Mammalian Erythrocytes Plus Heat Stability Test and Use of Agglutination of Mammalian Erythrocytes Plus Erythrocyte Elution Time to Characterize Isolates of Newcastle Disease Virus

Maduike C. O. Ezeibe, John A. O. Okoye, Ihuoma E. Ezeala, Ijeoma J. Mbuko, Augustine A. Ngene

Microbiology Research Journal International, Page 730-736
DOI: 10.9734/BMRJ/2014/7128

Aim: For a simpler, more rapid and more accurate method of characterizing  new isolates of Newcastle disease virus (NDV), agglutination of mammalian erythrocytes (HA)  plus heat stability  test  and  agglutination of  mammalian erythrocytes  plus  erythrocytes  elution time (EET)  were compared with  use of Intra-cerebral pathogenicity index (ICPI) to characterize the isolates. 

Materials and Methods: NDV isolates characterized by their ICPI were re-characterized by HA of mammalian erythrocytes plus heat stability test and by HA of mammalian erythrocytes plus EET. Rate of agreement of each of the two combinations with ICPI was calculated.

Results: HA of mammalian erythrocytes plus heat stability agreed with ICPI in characterizing 10 of the 12 NDV isolates (83.3%) while use of HA of mammalian erythrocytes plus EET agreed with ICPI in all the 12 isolates (100%).

Conclusion: It was concluded  that  use of combination of  agglutination  of mammalian erythrocytes  and  EET to characterize NDV isolates  has better agreement with use of ICPI to characterize the virus  than use of combination of agglutination of mammalian erythrocytes  and  heat stability test.

Open Access Original Research Article

Role of Different Additives on Survival of Serratia plymuthica HRO-C48 on Oilseed Rape Seeds and Control of Phoma lingam

Ruba Abuamsha, Mazen Salman, Ralf-Udo Ehlers

Microbiology Research Journal International, Page 737-748
DOI: 10.9734/BMRJ/2014/8390

For optimum inhibition of plant pathogens, biocontrol agents must be maintained at higher density and survive for a long time in the plant system. Biopriming of seeds with bacteria in the presence of different additives provide a promising technique that might improve the efficacy of biocontrol agents and their application. The aim of this work was to test different additives and stickers on the activity of Serratia plymuthica in oilseed rape in controlling Phoma lingam. Seeds were soaked in bacterial suspensions (log10 11 CFU ml-1) containing one of the following Stickers and additives in a ratio of 1:1:1 (w:v:v): Sodium alginate, Dextran T 40, Polyvinyl alcohol, Methylcellulose, Gum Arabic, Raffinose, Tween 20, and Paraffin oil. Seeds were stored either at room temperature or at 4°C. Number of bacteria inside the seeds was monitored over a period of 12 months. The effect of additives and stickers on bacterial efficacy in controlling Phoma lingam was evaluated in pot experiments. Number of bacterial cells inside the seeds was significantly higher in the seeds coated with GA+MgSO4, RF+MC and PA (log10 7.5 ± 0.2, log10 7.5 ± 0.2 and log10 7.4 ± 0.4, respectively). Interestingly, bacterial concentration in seeds stored at 4°C was higher than that in seeds stored at 20°C. Moreover, after storage for 8 months, Serratia plymuthica was able to control the black leg disease. Our results showed that some additives and stickers prolonged the shelf life of bacteria inside the seeds and improved the efficacy of bacteria in controlling the disease.

Open Access Original Research Article

Comparative Study of the Antibacterial Effect of Mouth Washes and Vernonia amygdalina (del.) on Some Tooth Decay Causing Bacteria

B. J. Akinyele, B. O. Oladejo, A. I. Akinyemi, L. O. Ezem

Microbiology Research Journal International, Page 749-758
DOI: 10.9734/BMRJ/2014/6386

Aims: To determine the antibacterial effect of the ethanol stem extract of Vernonia amygdalina (bitter-leaf) and some mouth washes against some bacteria that have been implicated in causing tooth decay so as to establish the role of herbal medicine and chemical compounds in oral hygiene.

Study Design: In vitro assay of antibacterial activities

Place and Duration of Study: Dental Department of the State Specialist Hospital, Akure, Ondo State, Nigeria and Department of Microbiology, Federal University of Technology, Akure, Nigeria, between October, 2012 and January, 2013.

Methodology: Bacterial isolates were collected, identified, standardized and the stem extract was prepared. Phytochemical screening of the extracts was carried out as well as the in vitro antibacterial assay using agar well diffusion technique. Minimum Inhibitory Concentration and antibiotics sensitivity test (disc diffusion assay) were also determined.

Results: The stem extract showed the presence of anthraquinone, alkaloid, saponin, steroid and cardiac glycoside. The ethanolic stem extract of Vernonia amygdalina inhibited all the test isolates at a concentration of 50 mg/ml with the highest zone of inhibition observed against Staphylococcus aureus (26.0 mm) while the least zone of inhibition of 14.0 mm was observed against Streptococcus mutans. Colgate mouthwash exhibited the highest zone of inhibition against Staphylococcus aureus while the least was recorded by Brett against Staphylococcus epidermidis. The antibacterial assay compared well with Ciprofloxacin, and in most cases higher zones of inhibition were recorded than the commercial antibiotics. The Minimum Inhibitory Concentration of the mouth washes ranged from 30 to 70% while it was 12.5 mg/ml for the stem extract.

Conclusion: Bioactive components of Vernonia amygdalina can be incorporated as ingredients in manufacturing mouthwashes and the plants’ stem can be used in the form of chewing stick. Further purification of the extract is necessary to further enhance greater antibacterial activity.

Open Access Original Research Article

Isolation and Molecular Biological Characterization of Fowlpox Virus from Specimen of Cutaneous Nodular Lesions from Chickens in Several Regions of Tanzania

S. N. Masola, A. Mzula, H. N. Tuntufye, C. J. Kasanga, P. N. Wambura

Microbiology Research Journal International, Page 759-771
DOI: 10.9734/BMRJ/2014/8420

Aim: To investigate molecular and evolutionary characteristics of genes of fowlpox virus (FWPV) isolates from chickens in Tanzania.

Study Design:  Experimental.

Place and Duration of Study: Faculty of Veterinary Medicine, Sokoine University of Agriculture, Morogoro, Tanzania; between November 2011 and October 2013.

Methodology: Samples of cutaneous nodular lesions were collected from featherless parts of chickens (n = 154) suspected to have fowl pox in 14 regions of Tanzania followed by virus isolation, DNA extraction, polymerase chain reaction (PCR) amplification of the P4b gene, gel electrophoresis of PCR products, purification of PCR products, sequencing of purified PCR products and finally analysis of sequence data using standard procedures.

Results: The disease was confirmed in 12 regions, out of 154 investigated samples 66 (42.86%) were found to contain FWPV, indicating that the 66 chickens from which the samples were collected had fowl pox as a result FWPV infection. Sequence analysis revealed that the Tanzanian FWPV isolates were 99.65 – 100% identical to each other and 99 – 100% identical to several published sequences of FWPV isolates from various countries in different continents of the world, including Europe and Asia. Phylogenetic analysis revealed that all Tanzanian isolates belong to clade A, subclade A1.

Conclusion: Based on the findings of this study it is concluded that currently fowl pox is prevalent in several regions of Tanzania, caused by FWPVs which are genetically and phylogenetically closely related. However, these findings do not rule out the possibility of existence of genetic divergence among FWPVs currently prevalent in Tanzania. In order to rule out or detect genetic divergence (if any) among FWPVs currently prevalent in the country, other studies aimed at investigating molecular and evolutionary characteristics of genes in other genomic regions are highly recommended.

Open Access Original Research Article

Comparative Microbial Assessment of Fermented Lima Bean (Phaseolus lunatus) and Locust Bean (Parkia biglobossa) in Production of Daddawa

Elizabeth Oluremi Farinde, Hezekiah Adekanmi Adeniran, Sumbo Henrietta Abiose

Microbiology Research Journal International, Page 772-784
DOI: 10.9734/BMRJ/2014/7514

Aim: Daddawa is a condiment traditionally produced from fermentation of locust bean (Parkia biglobossa).This study is directed at exploitation of lima bean–an underutilized but cultivated legume for production of daddawa analogue-a popular West African food condiment. Microbiological characteristics of the two condiments are compared.
Methodology: Lima bean and locust bean seeds were separately heat processed and fermented into daddawa analogue and daddawa respectively. The microbial load, microbial types and their succession in fermented samples were monitored over a fermentation period of 72h.
Results: Results showed that total viable bacteria count increased from 4.74 to 9.25 and 5.87 to 8.10 (log CFUg-1) within 72h of fermentation in fermenting lima bean and locust bean respectively. The bacteria isolates from the fermenting lima bean and locust bean were identified as Bacillus subtilis, B. licheniformis,B. brevisB. coagulansB. pumilus and B. megaterium.
B. licheniformis occurred virtually throughout the fermentation period in the tested legumes while B. subtilis and B. pumilus were isolated from 24th and 36th h till the end of fermentation in the fermented lima bean samples respectively.
Conclusions: The study has established that daddawa produced from the two legumes have similar microbiological characteristics. Production of daddawa from locust bean has not been commercialized to date as result of unreliable source of raw material. Locust bean is presently obtained from the wild. Lima bean which is cultivated could be a sustainable raw material for this food condiment which is popular in many West African communities.

Open Access Original Research Article

Clinical and Diagnostic Significance of Methicillin-resistant Staphylococcus aureus Infection in Bangladesh: A Systematic Review

M Abdullah Yusuf, AFM Arshedi Sattar, Zakir Husain Habib, Sushmita Roy

Microbiology Research Journal International, Page 785-797
DOI: 10.9734/BMRJ/2014/8071

Background: Accurate diagnosis of Methicillin-resistant Staphylococcus aureus (MRSA) is essential for the clinician. In Bangladesh MRSA creates a great problem for the treatment of infection.

Objective: The purpose of the present study was to observe the clinical and diagnostic significance of MRSA infection in Bangladesh.

Design: Systematic review of published articles in Bangladesh.

Data Sources: PubMed (Medline), Embase, Scopus, Web of Science, the Cochrane Library, and the World Health Organization (WHO) Regional Databases (African, eastern Mediterranean, Latin American and Caribbean, western Pacific, and southeast Asian regions) as well as Google Scholar, Banglajol, Asiajol.

Review Methods: The search was restricted to full articles published from January 2000 (publication date of the first study identified by the research) to December 2013. Studies were excluded that did not provide appropriate data on the prevalence of MRSA. Only English language was applied.

Result: A total number of 125 studies were identified during systematic review which were relevant to the present research question and among these only 14 studies were met the criteria for analysis. The level of evidence and freedom from bias of these studies were generally low. MRSA was diagnosed phenotypic in most of the articles. Majority were isolated from skin wound. The isolation rate of MRSA among all culture isolates ranged from 4.8-78.7%. From all studies diagnosis of MRSA infection was done from hospital setting; however, only two studies had been reported from community settings though the CDC definition was not followed in either study.

Conclusion: Significance of methicillin-resistant Staphylococcus aureus infection in Bangladesh is very high leading to a huge clinical as well as laboratory burden in the heath care facilities as well as in the community settings of Bangladesh.

Open Access Original Research Article

Effect of Incubation Temperature and Human Serum on Yeast to Hyphal Morphogenesis in Vaginal Candida albicans and its Correlation to Virulence Markers

Talat A. El-Kersh, Raid A. Alakeel, Abdulaziz H. Alhamidi, Ziab Z. Al-Ahmadey

Microbiology Research Journal International, Page 798-812
DOI: 10.9734/BMRJ/2014/8822

Aims: To investigate the effect of human serum, starvation and /or variation in incubation temperature on yeast and pseudo-hyphae and/ or hyphal cell differentiation in vaginal Candida albicans strains and, its correlation to exoenzymes productivity.

Study Design: A total of 31 C. albicans strains previously isolated from high vaginal swab specimens of pregnant Saudi women, as well as the C. albicans QC strain ATCC 10231 were recruited from Brain Heart Infusion-glycerol stock cultures(-80ºC) & included in the study.

Place and Duration of Study: Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud University, Riyadh, Saudi Arabia, between  September 2013 and December 2013.

Methodology: Each of thirty one vaginal C. albicans strains and the QC strain (ATCC 10231)  was grown in Modified Sabouraud Broth (MSB) at 25ºC and at 37ºC with or without addition of  20  %  human  serum;  and  morphological  growth  was  observed  at  2  hours intervals by phase contrast microscopy. Selected C. albicans strains that showed ability and/or weak-ability of yeast-hyphal transition were also tested for their exo-hydrolytic enzymes of phospholipase, and proteinase as caseinase, & gelatinase, and coagulase, virulence markers.

Results: Showed that at 25ºC 28/31(90.3%) strains were non filamentous, 3/31(9.6%) strains were moderately filamentous, and 0.0% strong filamentous, in comparison, at 37ºC those numbers were 19/31(61.3%), 10/31(32.3%), and 2/31 (6.4%) respectively, suggesting that mere increase in temperature from 25ºC to 37ºC remarkably increases yeast morphogenesis to filamentous forms. Such increase was significantly (P<0.001) more pronounced upon the addition of 20% serum at either incubation temperature of 25ºC or 37ºC as expected. Generally the presence of serum and/or incubation at high temperature (37ºC),  speeds off hyphal growth formation. Additionally, results also showed that 8/31 (25.8%) strains exhibited transition to hyphal forms only in presence of serum, whereas 7/31 (22.6%), apparently lost their capacity to switch to hyphal forms even in presence of serum and/or at temperature of 37ºC incubation. In contrast three strains 3/31, (9.7%) expressed such ability of filamentous growth in presence or absence of serum at 37ºC as well as 25ºC. These strains also showed enhanced secretion of exoenzymes. Therefore, these strains would be the most virulent ones. Whereas those strains (7/31, 22.6%) that did not show filamentous growth at any of the examined growth conditions would be considered as less virulent strains. However, considering the limited number of strains tested in this study, these findings require further substantiation by large sample size and in vivo animal studies.

Conclusion: Results obtained suggest that vaginal C. albicans strains are heterogenous in their potency to switch from yeast to hyphae.  Strains which show morphogenesis in absence of serum and/or at low temperature (25ºC) exhibit higher exoenzymes activity suggesting that these strains are more pathogenic

Open Access Original Research Article

The Bacteriology of Discharging Ear (Otitis Media) Amongst Children in Owerri, Imo State, South East Nigeria

N. U. Nwogwugwu, I. N. S. Dozie, E. C. Chinakwe, I. N. Nwachukwu, N. N. Onyemekara

Microbiology Research Journal International, Page 813-820
DOI: 10.9734/BMRJ/2014/5304

The prevalence of causative agents of acute otitis media amongst children in some parts of Owerri, Imo State - Nigeria was investigated, between the months of September and December, 2012 using standard microbiological methods. One hundred and fifty two swabs from ear discharge were collected from children under 12 years of age. The results revealed that 128 (84.2%) were positive for bacterial growth while 24 (15.8%) had no growth. The predominant organisms isolated included Pseudomonas aeruginosa (37.5%), Staphylococcus aureus (23.4%), Proteus species (12.5%), Klebsiella pneumoniae (7.8%), Escherichia coli (7.8%) and Streptococcus pneumoniae (7.8%). Antibiotic susceptibility tests revealed that all the isolates were multiresistant to six or more of the tested antibiotics while the most prevalent organisms were susceptible to gentamycin, chloramphenicol and cotrimoxazole. Pseudomonas aeruginosa and Proteus mirabilis were susceptible to ofloxacin and ciprofloxacin. All the isolates were resistant to amoxicilline/clavulanate spectinomycin and ofloxacin (except Staphylococcus aureus). Therefore, chloramphenicol, gentamycin, cotrimoxazole ciprofloxacin and ofloxacin, are suggested as topical treatment in the management of acute otitis media and good personal hygiene is also encouraged.

Open Access Original Research Article

An Improved Protocol for Extraction of Metagenomic DNA from High Humus, Alkaline and Saline Soil of Chinampa for T-RFLP Fingerprinting Analysis

Salvador Embarcadero-Jiménez, Feng Long Yang, Raquel Freye-Hernández, Yanelly Trujillo-Cabrera, Flor N. Rivera Orduña, Hong Li Yuan, En Tao Wang

Microbiology Research Journal International, Page 821-830
DOI: 10.9734/BMRJ/2014/9670

Aims: The principle aim of this study was to obtain high quality metagenomic DNA from the high humus-containing, alkaline soils of the chinampas, an artificial sustainable agro-ecosystem.

Study Design: Different protocols reported previously were tested and were modified to extract the metagenomic DNA. Quality of the DNA samples was evaluated by amplification of 16S rRNA gene with PCR and T-RFLP (Terminal Restriction Fragment Length Polymorphism) analysis.

Place and Duration of Study: This study was performed in Department of Microbiology, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional during 2011-2012.

Methodology: Four soil samples were collected from two chinampas at the depth of 0-30 cm and 30-60cm. A protocol started with repeated prewashing before the direct cell lysis with lysozyme followed by SDS treatments, frozen and melting cycling was developed which combined the DNA isolation and purification procedures. The 16SrRNA genes were amplified from the extracted metagenomic DNAs and were used for T-RFLP fingerprinting analysis.

Results: The 16SrRNA genes were amplified from all the DNA extracts corresponding to the four soil samples and were successfully used in the T-RFLP analysis, which generated 25 to 109T-RFs in the four soil samples digested separately with the restriction endonucleases HaeIII, HhaI and MspI.

Conclusion: The protocol developed in the present study could generate high molecular weight and high quality metagenomic DNA from soils with high content of humic materials, for which the other reported protocols were not functioned. This soil harboured very diverse and unique bacterial communities belonging to at least nine phyla that might contribute to the high soil fertility.