Open Access Original Research Article

Development of Antibiotic Resistance in Herbal Drug-sensitized Staphylococcus aureus Isolate

Monsi Tombari Pius, Wokem Gloria Ngozika, Aleruchi Promise Chizi

Journal of Advances in Microbiology, Page 1-7
DOI: 10.9734/JAMB/2017/37893

Background: Herbal remedies are locally produced drugs usually made from plant sources. Due to their inexpensive nature, low-income individuals prefer them over modern antibiotics in the treatment of infections.

Aims: The purpose of the current study was to determine the ability of a herbal drug called Goko Alcoholic Bitters (GAB) to induce resistance in opportunistic pathogens such as Staphylococcus aureus.

Methods: To evaluate the antimicrobial efficacy of GAB on S. aureus, Disc diffusion method was employed to observe the zones of clearance by GAB. Overnight cultures of S. aureus grown in Tryptone Soy Broth (TSB) at an optical density of 0.5 were serially diluted to 109. Five (5) different concentrations of GAB (0.018 g/ml, 0.006 g/ml, 0.002 g/ml, 0.0007 g/ml and 0.0002 g/ml) were added to the serially diluted culture. The growth responses of these different dilutions were checked against S. aureus isolate in vitro. The experiment was monitored for 24 and 48 hrs conditions using two different growth determination methods; turbidity and colony forming unit counting methods. The five (5) different conditions were further analyzed by plating 200 µl of bacterial broth on Tryptic Soy Agar (TSA) and antibiotic sensitivity discs were embedded onto the agar. 200 µl of control bacterial Tryptone Soy broth containing S. aureus not exposed to the herbal drug was also performed alongside for comparison. This was repeated for both 24 and 48 hrs exposure to GAB.

Results: The efficacy of GAB showed some levels of antimicrobial activity, however, were significantly lower compared to Ampicillin (10 µg). The growth response analysis showed gradual reduction but not complete elimination with the highest concentration of GAB. GAB-sensitized S. aureus showed resistance seen in the reduced zones of clearance to some Septrin 10 µg (SEP), Ampicillin 10 µg (AMP) and Nalidixic acid 10 µg (NA) which were effective against untreated S. aureus (control).    

Conclusion: GAB has some levels of antimicrobial efficacy but antibiotic resistance gets induced in the case of surviving GAB-pretreated S. aureus.

Open Access Original Research Article

Isolation and Molecular Characterization of a Wild Type B. cinerea from Infected Bottle Gourd (Lagenaria sicerari) in China

M. Kamaruzzaman, Ali Akbar Bhuiyan, Mohammad Omar Faruque

Journal of Advances in Microbiology, Page 1-10
DOI: 10.9734/JAMB/2017/37110

Botrytis cinerea is an important phytopathogenic fungus that causes diseases in wide range of hosts, including various fruits, vegetables and ornamental plants. In this study, infected bottle gourd was collected from the commercial farm of Wuhan, China to characterize the pathogen. Morphological identification was done based on colony characteristics, shape and size of conidia, conidiophores and formation of sclerotia. Results showed that conidiophores arise straight from the mycelia with length of 2-8 µm × 220-480 µm, an average conidial dimension of 5-8.5 μm × 9-13 μm, and a size of the sclerotia ranging from 0.7-4.5 mm × 1 – 8.5 mm. Species specific primers, Bc-f /Bc-r and Bc108+/Bc563- produced 354 and 450 bp fragments, respectively. Sequence of PCR amplification of the rDNA- ITS region produced a 547 bp fragment using the universal primer pair ITS1/ITS4. In the phylogenetic tree based on ITS nucleotide sequences, the representative isolate was located within a clade comprising reference isolates of B. cinerea. Strain XT5-2 germ tubes formed infection cushion on heat-killed onion epidermis and successfully entered into the epidermal cells. In this study, it was found that B. cinerea grew well at 20°C rather than below 15°C and above 25°C, respectively. Pathogenicity assays showed B. cinerea strain was virulent and displayed water-soaked lesions after 3 days. Based on morphological symptoms, mycological features, molecular analysis and pathogenicity bio assay, this fungus was identified as B. cinerea. To the best of our knowledge, this is the first study on bottle gourd (Lagenaria siceraria (Molina) Standl.) as a new host of B. cinerea in China.

Open Access Original Research Article

Effects of Ampicillin on the Gut Microbiome of an Adult Male as Determined by 16S rRNA V4 Metagenomics Sequencing and Greengenes Bioinformatics Suite

Kingsley Anukam

Journal of Advances in Microbiology, Page 1-18
DOI: 10.9734/JAMB/2017/38867

Aims: To determine the effects of ampicillin on the relative abundance of the gut microbiota using Next Generation Sequencing (NGS) metagenomics platform and to predict metabolic functions with bioinformatics suite.

Methods: The subject who has taken no antibiotics for the past six months provided fecal sample on day 0 (pre-antibiotics), day 10 (antibiotics) to treat sour throat infection and after 90 days (post-antibiotics). DNA was extracted, metagenomics library prepared and 16S rRNA V4 region was amplified using custom bar-coded primers before sequencing with IlluminaMiseq program. Sequence reads were analyzed with IlluminaBasespace algorithm and greengenes bioinformatics suites. Metabolic functional prediction was accomplished using the Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) algorithm.

Results: Pre-antibiotics sequence reads were classified into 20 phyla, 35 Class, 71 Order, 147 Family, 275 Genera, and 401 Species-level categories. Ampicillin significantly influenced the gut microbiota as the sequence reads were reduced to 8 phyla, 15 Class, 24 Order, 41 Family, 72 Genera, and 98 Species-level categories. Post-antibiotics sample (90 days after antibiotic treatment) generated sequence reads classified into 17 phyla, 28 Class, 60 Order, 120 Family, 221 Genera, and 313 Species-level categories.

Conclusion: Ampicillin reduced the diversity of the core bacterial phylogenetic taxa with a corresponding increase in Firmicutes-Bacteroidetes ratio from 2.4:1 (pre-antibiotics) to 6.5:1 (antibiotics). A high proportion of Veillonella species were observed during ampicillin intake. Some bacterial metabolic functions such as carbohydrate (Ascorbate and aldarate), amino acid (D-Arginine and D-ornithine) and vitamin (pantothenate and CoA biosynthesis) metabolisms were stimulated by ampicillin in the subject.

Open Access Original Research Article

Control of Cowpea (Vigna unguiculata L. Walp) Diseases with Intercropping of Maize (Zea mays L) and Spray of Plant Extracts

M. J. Falade, O. A. Enikuomehin, O. A. Borisade, M. Aluko

Journal of Advances in Microbiology, Page 1-10
DOI: 10.9734/JAMB/2017/38156

Field experiment was conducted to evaluate the effect foliar spray of aqueous extracts of Datura stramonium, Ricinus communis and Jatropha gossypifolia at three concentrations (65 50 and 30%) on cowpea anthracnose disease caused by Colletotrichum lindemuthianum in sole cowpea and cowpea / maize intercrop. The total area of the farm was 270m2 and cowpea population in each cropping system was 324 stands at a spacing of 60x30 cm while the population of maize in the cowpea/maize intercrop was 162 stands at a spacing of 90x60 cm. Spraying of the plant extracts started two weeks after planting (WAP) and continued until 8 WAP. The extract of D. stramonium reduced the incidence and severity of the disease most compared to other extracts. Similarly, the disease incidence in cowpea/ maize intercrop was lower than in sole cowpea. The disease incidence at 65, 50 and 30% concentrations on pooled mean for the sole crop were 11, 15 and 21% whereas that of intercrop plants were 9, 12 and 16%  respectively. The percentage of normal seeds obtained in the study was concentration dependent being highest at 65% in all the extracts and was higher in intercropped (95%) than the sole crop (93%).Similarly, fungal infection on seeds was lower at all concentrations compared to the control. The study therefore shows that extracts of the plants and intercropping of non-host reduced incidence and severity of anthracnose infection with corresponding increase in yield.

Open Access Review Article

The Role of Nanotechnology in Food Industries- A Review

I. E. Aigbogun, S. S. D. Mohammed, A. A. Orukotan, J. D. Tanko

Journal of Advances in Microbiology, Page 1-9
DOI: 10.9734/JAMB/2017/38175

This paper, points on the synoptic roles of nanotechnology in food industries which cover sectors such as food processing, encapsulation of nutraceuticals, food packaging, and food quality monitoring to enhance food safety and security. Nanotechnology refers to the engineering of material structures and sizes ranging from 1–100 nm to suit the need for different sectors. Nanotechnology possesses great potential to apply at all areas of the food sector, ensuring modification of color and flavor, nutritional values, increasing the shelf life of food, and monitoring the integrity of food via barcodes such as cold chain, i.e. whenever there is a slight change occur in food storage condition because of its submicroscopic nature. It can also modify permeation of materials by the incorporation of synthesized nanoparticles (zinc, silver, gold e.t.c) for better packaging system. From this review, it is evidently clear that the role of nanotechnology in food industries cannot be overemphasized. Currently, in nanotechnology, nanosensors serve as diagnostic devices to monitor food processes to meet the wholeness and safety of food.