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Article

Characterization of rpsL Gene Mutations in Streptomycin-Resistant Mycobacterium tuberculosis Isolates

1Department of Microbiology, Intermediate Reference Laboratory, Government Hospital for Chest Diseases, Puducherry, India


American Journal of Microbiological Research. 2014, 2(3), 80-85
DOI: 10.12691/ajmr-2-3-1
Copyright © 2014 Science and Education Publishing

Cite this paper:
Usharani Brammacharry, Muthuraj Muthaiah. Characterization of rpsL Gene Mutations in Streptomycin-Resistant Mycobacterium tuberculosis Isolates. American Journal of Microbiological Research. 2014; 2(3):80-85. doi: 10.12691/ajmr-2-3-1.

Correspondence to: Muthuraj  Muthaiah, Department of Microbiology, Intermediate Reference Laboratory, Government Hospital for Chest Diseases, Puducherry, India. Email: muthuraj1970@gmail.com

Abstract

The worldwide emergence and spread of drug - resistant strains of Mycobacterium tuberculosis has adverse effects on tuberculosis (TB) control programs. The goal of this paper is to describe the advances made in the understanding of the molecular basis of M. tuberculosis resistance to streptomycin and to discuss the potential of molecular methods in early diagnosis of streptomycin resistant TB.Molecular methods such as DNA sequencing, polymerase chain reaction have been used to identify/detect mutations in rpsL gene-encoding proteins.Of the 77 SM resistant isolates, 22 (28.6%) exhibited mutation at codon 43 Lys→Arg, 11 (14.3%) isolates exhibited mutation at codon 83 Arg→Gln, 9(%) isolates exhibited mutation at codon 95 Tyr→His,whereas remaining 23 (40.8%) SM resistant isolates showed no mutation in rpsL gene. Among the 77 SM resistant isolates, 31(40.2%) were SM mono resistant and 46(59.8%) strains were poly resistant. All the 23 SM susceptible isolates as well as reference strain M.tuberculosis H37Rv exhibited wild type sequences of rpsL gene. Molecular methods to detect the most frequent mutations in the gene encoding functions that are targets for streptomycin drug have provided encouraging results for early diagnosis of resistance nature.

Keywords

References

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Article

Evaluation of New Bacteriocin as a Potential Short-Term Preservative for Goat Skin

1Leather Processing Division, Central Leather Research Institute, Adyar, Chennai, India

2CHORD, Central Leather Research Institute, Adyar, Chennai, India


American Journal of Microbiological Research. 2014, 2(3), 86-93
DOI: 10.12691/ajmr-2-3-2
Copyright © 2014 Science and Education Publishing

Cite this paper:
J. Kanagaraj, A. Tamil Selvi, T. Senthilvelan, N.K. Chandra Babu, B. Chandrasekar. Evaluation of New Bacteriocin as a Potential Short-Term Preservative for Goat Skin. American Journal of Microbiological Research. 2014; 2(3):86-93. doi: 10.12691/ajmr-2-3-2.

Correspondence to: T.  Senthilvelan, Leather Processing Division, Central Leather Research Institute, Adyar, Chennai, India. Email: mtsenthil85@yahoo.com

Abstract

Common salt (sodium chloride) and chemical preservatives are used for preservation of skins and leather to protect from microbial attacks, which leads to severe pollution problem. Bacteriocin is an eco-friendly option for skin and leather preservation in leather industry, which contains antimicrobial and antifungal components their fluid. Bacteriocin was extracted from Lactobacillus plantarum isolated from chicken meat by submerged fermentation method. The bacteriocin was tested against the skin putrefying organism Pseudomonas aeruginosa and Bacillus putrefaciens by plate assay method, which showed the activity of 200 AU/ml for Pseudomonas aeruginosa, and 340AU/ml for Bacillus putrefaciens. These two cultures were further analysed by SEM, which showed cleavage of bacterial cell walls due to action of bacteriocin. This study revealed the presence of antimicrobial activity in bacteriocin. Then the bacteriocin was tested for preservation of goat skin infected by skin putrefying organisms of Pseudomonas aeruginosa and Bacillus putrefaciens, and its efficacy was studied. The microbial infected goat skin was preserved with 15% bacteriocin solution and stored for seven days at room temperature, which showed the complete inhibition of microbial growth even after seven days of storage. Then the goat skin was further processed into crust leather and which was analysed by SEM and physical testing data. The SEM analysis and physical testing data revealed no significant changes in the fiber structure of the skin due to preservation by bacteriocin. The pollution load generation (Total Dissolved solvents) has reduced at the level of 94.4% and 95.6% (of chlorides) during soaking process. The overall experiments revealed that the bacteriocin can be used as potential bio preservatives for preservation of skin and leather.

Keywords

References

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Article

Bio-signature of Ultraviolet-Radiation-Resistant Extremophiles from Elevated Land

1Division of Biological and Health Sciences, University of Pittsburgh, Bradford, PA, USA

2Department of Environmental Hydrology and Microbiology, Ben-Gurion University of the Negev, Sede-Boquer Campus, Israel


American Journal of Microbiological Research. 2014, 2(3), 94-104
DOI: 10.12691/ajmr-2-3-3
Copyright © 2014 Science and Education Publishing

Cite this paper:
Prashant Gabani, Dhan Prakash, Om V. Singh. Bio-signature of Ultraviolet-Radiation-Resistant Extremophiles from Elevated Land. American Journal of Microbiological Research. 2014; 2(3):94-104. doi: 10.12691/ajmr-2-3-3.

Correspondence to: Om  V. Singh, Division of Biological and Health Sciences, University of Pittsburgh, Bradford, PA, USA. Email: ovs11@pitt.edu, ovs11@yahoo.com

Abstract

Microorganisms with the ability to survive high doses of radiation are known as radiation-resistant extremophiles. This study attempts to demonstrate the diversity of microorganisms resistant to ultraviolet radiation (UVR) in the natural environment in order to investigate the molecular and physiological mechanisms by which these microorganisms survive under extreme radiation. We hypothesized that topsoil from elevated land (hills) would reveal a diverse variety of UVR-resistant extremophiles with modulated proteins/enzymes. A total of 10 different UV-C (UV subtype-C)-resistant extremophiles—UVP1, UVP3, UVP4, UVR1, UVR3, UVR4, UVR5a, UV20hr, YLP1, and BR2—were isolated and identified using 16S rRNA sequences for nearest homologues. All the isolates showed prolonged resistance against UV-C: 3.44 x 105 - 2.74 x 106 J/m2. Phylogenetic analysis between and within the UVR isolates revealed their relationship with other soil microorganisms using different outgroups. A unique pattern of protein expression at 25-50kDa was observed on SDS-PAGE under UVR and non-UVR from six prominent UVR isolates. Current studies are finding extreme UV-C-resistant in naturally occurring microorganisms found in stress-free environments.

Keywords

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Article

Phosphate Solubilizing Rhizobia Isolated from Vigna trilobata

1Department of Botany & Microbiology, Acharaya Nagarjuna University, Guntur, India


American Journal of Microbiological Research. 2014, 2(3), 105-109
DOI: 10.12691/ajmr-2-3-4
Copyright © 2014 Science and Education Publishing

Cite this paper:
G. Kranthi Kumar, M. Raghu Ram. Phosphate Solubilizing Rhizobia Isolated from Vigna trilobata. American Journal of Microbiological Research. 2014; 2(3):105-109. doi: 10.12691/ajmr-2-3-4.

Correspondence to: M.  Raghu Ram, Department of Botany & Microbiology, Acharaya Nagarjuna University, Guntur, India. Email: mraghuram2002@gmail.com

Abstract

Rhizobial strains were isolated from root nodules of Vigna trilobata plants raised in soils of different districts of Andhra Pradesh, India. Among the 23 strains of rhizobia isolated, 6 strains were proved to be positive for phosphate solubilization. The strains were identified as Sinorhizobium sp. strain MRR101-KC428651, Agrobacterium tumefaciens strain MRR102- KC428652, Rhizobium sp. strain 103 – JX576499 ; Sinorhizobium kostiense strain MRR104- KC428653; Agrobacterium tumefaciens strain MRR105- KC428654 and Rhizobium sp. strain MRR106- KC428655 after 16S rDNA sequencing. S. kostiense strain MRR104 was found to be better than strains of Agrobacterium tumefaciens and Rhizobium sp. in phosphate solubilization with maximum zone of solubilization (18mm) on standard Pikovskaya’s medium and with maximum P2O5 liberation of 510 µg/ml in liquid medium. The optimization for maximum phosphate solubilization was done by using different carbon and nitrogen sources. Glucose was preferred as carbon source by all the strains studied, with 10 fold increase in solubilization of phosphorous than other carbon sources. Phosphate solubilization increased with increase in concentration of glucose up to 3% in all the strains studied. Some strains preferred ammonium sulphate and others preferred nitrates as nitrogen source for phosphate solubilzation, indicating that strains of V. trilobata are adopting two different mechanisms for solubilization. Reduction in pH with increased phosphate solubilization efficiency was also observed among the strains, irrespective of the carbon sources tested. Strain S. kositense MRR104 was proved to the better strain with maximum libration of phosphorous along with maximum reduction in pH as 5.13. Sinorhizobium strains performed better than that of Rhizobium and Agrobacterium in solubilization of phosphorous.

Keywords

References

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Article

Nasal Carriage of Staphylococcus in Health Care Workers in Benghazi Hospitals

1Department of Laboratory, Eye Hospital, Benghazi, Libya

2Department of Botany, Faculty of Science, Benghazi University, Libya


American Journal of Microbiological Research. 2014, 2(4), 110-112
DOI: 10.12691/ajmr-2-4-1
Copyright © 2014 Science and Education Publishing

Cite this paper:
Nadia. E. Al-Abdli, Saleh.H. Baiu. Nasal Carriage of Staphylococcus in Health Care Workers in Benghazi Hospitals. American Journal of Microbiological Research. 2014; 2(4):110-112. doi: 10.12691/ajmr-2-4-1.

Correspondence to: Nadia.  E. Al-Abdli, Department of Laboratory, Eye Hospital, Benghazi, Libya. Email: batul.gr155@gmail.com

Abstract

This study was aimed to determine the frequency of staphylococcal nasal carriage of health care workers (HCWs) and antimicrobial susceptibility profile of the isolates in hospitals of Benghazi, Libya. The study was conducted in the period between April 2013 and August 2013 on HCWs from ten hospitals in Benghazi-Libya. Nasal swabs from anterior nares of HCWs were cultured and identified as Staphylococcus aureus (S. aureus), coagulase - negative staphylococci (CoNS) and methicillin-resistant S. aureus (MRSA) by using standard methods. Antimicrobial susceptibility testing was performed on Muller Hinton Agar using disc diffusion method. Of the 472 HCWs, (47.5%) were nasal carriers of S. aureus, (21.4%) for MRSA and (26.1%) for MSSA, (36.4%) of CoNS. The highest carriage rate for S. aureus was in Physician (30.6%) followed by nurses (20.4%), helpers (22.9%) and technicians (10.9%). Almost all of the isolates showed high resistance against penicillin (97.5%) and ampicillin (98.2%). The lowest resistance recorded was for augmentin (14.2%), gentamycin (7.1%), clindamycin (7.5%) ciprofloxacin (3.2%) and vancomycin (2.1%). Most of isolated strains were susceptible to rifampicin (98.9%). The emergence of resistant strains of S.aureus should be prevented by controlling the amount of antibiotics used in and out of hospitals. Also, better control measures of nosocomial infection must be established.

Keywords

References

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Article

Synergistic Bio-preservative Effects of Vernonia Amygdalina Leaves and Sacoglottis Gabonensis Stem Bark on Palm Wine from Elaeis Guineensis and Raphia Hookeri from Uturu, Nigeria

1Department of Microbiology, Michael Okpara University of Agriculture, Umudike, Abia State, Nigeria


American Journal of Microbiological Research. 2014, 2(4), 113-117
DOI: 10.12691/ajmr-2-4-2
Copyright © 2014 Science and Education Publishing

Cite this paper:
Onwuakor Chijioke .E, Ukaegbu-Obi K.M. Synergistic Bio-preservative Effects of Vernonia Amygdalina Leaves and Sacoglottis Gabonensis Stem Bark on Palm Wine from Elaeis Guineensis and Raphia Hookeri from Uturu, Nigeria. American Journal of Microbiological Research. 2014; 2(4):113-117. doi: 10.12691/ajmr-2-4-2.

Correspondence to: Onwuakor  Chijioke .E, Department of Microbiology, Michael Okpara University of Agriculture, Umudike, Abia State, Nigeria. Email: chijiokeonwuakor@gmail.com

Abstract

The synergistic bio-preservative effects of leaves and stem bark of Vernonia amygdalina and Sacoglottis gabonensis respectively on two (2) palm wine types namely, Elaeis guineensis and Raphia hookeri from Uturu-Nigeria was evaluated. The microbiological and biochemical changes of the palm wine brands were determined. R. hookeri brands were found to support more heterotrophic and coliform populations than the E. guineensis, while the later contained more yeast species. Identification of isolated species revealed the presence of Bacillus sp., Micrococcus sp., Lactobacillus sp., Brevibacterium sp. and Saccharomyces sp. from E. guineensis and R. hookeri. Moreso, heterotrophic counts and pH were observed to decrease as the fermentation time progressed. The combination of both V. amygdalina and S. gabonensis as preservatives lowered the bacterial and fungal load compared to the control and individual plant preservatives and reduced the rate of CO2 emission as well as keeping the pH fairly constant.

Keywords

References

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Article

Biotic Relationships: Distribution of Antibiotic Resistance Genes in Nosocomial Pathogens

1Department of Biological & Environmental Sciences, Troy University, Troy, AL 36082


American Journal of Microbiological Research. 2014, 2(4), 118-121
DOI: 10.12691/ajmr-2-4-3
Copyright © 2014 Science and Education Publishing

Cite this paper:
Tejpreet Chadha. Biotic Relationships: Distribution of Antibiotic Resistance Genes in Nosocomial Pathogens. American Journal of Microbiological Research. 2014; 2(4):118-121. doi: 10.12691/ajmr-2-4-3.

Correspondence to: Tejpreet  Chadha, Department of Biological & Environmental Sciences, Troy University, Troy, AL 36082. Email: tchadha@troy.edu

Abstract

The strong selective pressure exerted by excessive use of antibiotics in the last decades has increased the acquisition of resistance genes by horizontal gene transfer. Horizontal gene transfer contributed to the diversification of microorganisms by influencing traits such as metabolic potential of a bacterial cell, antibiotic resistance, symbiosis, fitness, and adaptation. The study of biotic relationships helped to examine how they may contribute to virulence such as transfer of toxin genes, antibiotic resistance genes. The current study examines the pattern of distribution of Ambler (molecular) classes of β-lactamases (A, B, C, D) that provides a basic understanding and an initial assessment of resistance genes that may have a different role in the natural environment in free living and symbiotic nosocomial pathogens. The distribution of the four different Ambler (molecular) classes of β-lactamases (A, B, C, and D) differs among different bacterial species. The results from absolute abundance data showed prevalence of class B β-lactamases was highest in free living and lowest in symbiotic bacterial species. Interestingly, class D β-lactamases was absent in symbiotic bacterial species. However, relative abundance class D β-lactamases was lowest for free-living bacterial species. The prevalence of class B β-lactamases based upon the total number of sequences checked predominated in pathogens that are free living when compared to other Ambler classes of β-lactamases. The study of biotic relationships helps to understand what selective or stimulatory pressures are driving the spread of antibiotic resistance genes. In the future, this may help to design effective strategies for preventing further increases in the incidence of antibiotic-resistant bacteria.

Keywords

References

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Article

Using Water with Oil Immersion Lens to Detect Malaria Parasite in Blood Film and Making a Comparison between Oil and Water Method

1Department of microbiology, Faculty of Medical Laboratory Science, The National Ribat University


American Journal of Microbiological Research. 2014, 2(4), 122-124
DOI: 10.12691/ajmr-2-4-4
Copyright © 2014 Science and Education Publishing

Cite this paper:
Hiba Siddig Ibrahim, Shamsoun Khamis Kafi. Using Water with Oil Immersion Lens to Detect Malaria Parasite in Blood Film and Making a Comparison between Oil and Water Method. American Journal of Microbiological Research. 2014; 2(4):122-124. doi: 10.12691/ajmr-2-4-4.

Correspondence to: Hiba  Siddig Ibrahim, Department of microbiology, Faculty of Medical Laboratory Science, The National Ribat University. Email: hibasiddig55@gmail.com

Abstract

Background: Oil immersion lens it is one of the microscope lenses that used in medical laboratory filed. It is used to magnify the smallest things and for detection of some important infectious diseases such as bacteria, parasite, anemia’s and bone marrow films, but uses lens alone without oil makes the image imperfect and unclear, so to make a better image scientists were used oil with these lenses (immersion oils lens) to avoid bending of light . Objectives: The main aim of this study is to be used distill water with oil immersion lens instead of synthetic oil to detect malaria parasite to low cost of using synthetic oil especially in faraway places due to difficulties of paying synthetic oil and also to be arrived to rural area, and so on to be used with other tests, beside avoiding using impurity oil due to the comment of D/ Fahad Awad (National Coordinator for Malaria Program), against using sesame oil especially in some laboratories at Al Gedaraf State, see Aray Alaam newspaper 5th December, 2013. Methodology: In this study a total of 200 subjects were included. A 3 drops from capillary blood samples were collected in clean dry dust free slides after disinfectant the 3ed or 4th finger for adult and the big toe hand or foot for child with 70% alcohol to make a thick blood films, or Aliquots of 2.5 ml of venous blood were collected by venous puncture after disinfectant the site of collection and the collected blood was drawn into EDITA containers to make thick blood films as above. The films were lets to dry by air, staining with Giemsa stain for 10 minute and then examined for the first times with DW with objective lens and for the second times with synthetic oil. Results: There were no differences between using oil or distill water (DW) for detection of malaria parasite by both techniques (100%), unless the high quality of oil image (100%), when comparing with distill water (DW) image (90%), due to variation in numerical aperture (NA) between both techniques. Conclusion: Based on the results of this study we can used distill water (DW) but with caution to detect malaria parasite by 90% when compared with oil 100% in case of poverty areas to low cost of using synthetic oil, and so on the image quality can arrives (99.5%) if do it in proper way.

Keywords

References

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Article

Effect of Varied Culture Conditions on Crude Supernatant (Bacteriocin) Production from Four Lactobacillus Species Isolated from Locally Fermented Maize (Ogi)

1Department of Microbiology, College of Natural Sciences, Michael Okpara University of Agriculture Umudike, Abia State, Nigeria

2Department of Microbiology, Faculty of Biological & Physical Sciences, Abia State University Uturu, Abia State, Nigeria

3Department of Product Development Programme, National Root Crop Research Institute Umudike, Abia State, Nigeria


American Journal of Microbiological Research. 2014, 2(5), 125-130
DOI: 10.12691/ajmr-2-5-1
Copyright © 2014 Science and Education Publishing

Cite this paper:
C.E Onwuakor, V.O Nwaugo, C.J Nnadi, J.M Emetole. Effect of Varied Culture Conditions on Crude Supernatant (Bacteriocin) Production from Four Lactobacillus Species Isolated from Locally Fermented Maize (Ogi). American Journal of Microbiological Research. 2014; 2(5):125-130. doi: 10.12691/ajmr-2-5-1.

Correspondence to: C.E  Onwuakor, Department of Microbiology, College of Natural Sciences, Michael Okpara University of Agriculture Umudike, Abia State, Nigeria. Email: chijiokeonwuakor@gmail.com

Abstract

This Lactic acid bacteria (LAB) predominates the micro flora of fermented products. They produce metabolites that inhibit the growth of food borne pathogens and spoilage microorganisms. The isolation and identification of LAB from fermented maize (Ogi) and the effect of varied culture conditions on crude supernatant production and activity was evaluated. Four (4) isolates of bacteriocin producing lactobacillus species (L. lactis, L. fermentum, L. casei and L. plantarum) with antibacterial activity against Salmonella typhimurium (ATCC 14028) and Shigella dysenteriae (ATCC 23351) were subjected to varying growth medium conditions. The crude supernatant production was tested at different physical and cultural conditions such as temperature (25, 30, 35 and 40°C), pH (5, 6, 7 and 8), sodium chloride (NaCl) concentration (2, 4, 6 and 8%) and incubation duration (12, 24, 48 and 72 hours). The optimum bacteriocin production judged by their different zones of inhibition of crude supernatant was recorded at temperature, 30°C and then 35°C. There were significant differences between all the incubation temperatures at P<0.05. Duration of incubation showed highest crude supernatant activity after 72 hours. Furthermore, optimal conditions for crude supernatant production were observed to be highest at pH 6.0 followed by 5.0 and then 2% NaCl concentration. There were significant differences between the zones of inhibition of crude supernatants produced against the indicator organisms at various media pH and salt concentrations at P<0.05. These crude supernatants may have a potential use in reducing contaminations during industrial processes, as food preservatives and may help in improving the gastro-intestinal tract by fighting off pathogenic bacteria.

Keywords

References

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Article

Enrichment of Activated Sludge Process in the Treatment of Industrial Waste Water

1Industrial Waste Water Research Laboratory, Division of Applied & Environmental Microbiology, Enviro Technology Limited, Ankleshwar, Gujarat, India


American Journal of Microbiological Research. 2014, 2(5), 131-137
DOI: 10.12691/ajmr-2-5-2
Copyright © 2014 Science and Education Publishing

Cite this paper:
Maulin P Shah. Enrichment of Activated Sludge Process in the Treatment of Industrial Waste Water. American Journal of Microbiological Research. 2014; 2(5):131-137. doi: 10.12691/ajmr-2-5-2.

Correspondence to: Maulin  P Shah, Industrial Waste Water Research Laboratory, Division of Applied & Environmental Microbiology, Enviro Technology Limited, Ankleshwar, Gujarat, India. Email: shahmp@uniphos.com

Abstract

In this paper, bioaugmentation was applied to upgrade a full-scale activated sludge system (A) into a contact oxidation system (B). Results showed that when chemical oxygen demand (COD) and ammonia nitrogen (NHþ4-N) concentration of the industrial wastewater were 320–530 mg/L and 8–25 mg/L, respectively, the bioaugmented process (A) took only 20 days when they were below 80 mg/L and 10 mg/L, respectively. However, the unbioaugmented conventional activated sludge process (B) spent 30 days to reach the similar effluent quality. As the organic loading rate (OLR) increased from 0.6 to 0.9 and finally up to 1.10 kg COD/m3 d, A showed strong resistance to shock loadings and restored after three days compared to the seven days required by B. Based on the results of this paper, it shows that bioaugementation application is feasible and efficient for the process upgrade due to the availability of the bioaugmented specialized consortia.

Keywords

References

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