Open Access Original Research Article

Antimicrobial Susceptibility Profiling of Staphylococcus aureus Isolates from Milk

W. Chaalal, H. Aggad, K. Zidane, N. Saidi, M. Kihal

Microbiology Research Journal International, Page 1-7
DOI: 10.9734/BMRJ/2016/24064

Aims: This study investigated the occurrence of Staphylococcus aureus in milk and its sensitivity to twenty antibiotics.

Study Design: The research was laboratory-based investigation.

Place and Duration of the Study: The study was carried out at the Laboratory of Hygiene and animal pathology, University of Tiaret, Algeria, between September 2012 and May 2013.

Methodology: Thirty eight milk specimens were collected from cattle and examined to estimate the prevalence of Staphylococcus aureus. The sensitivity of the isolates to twenty (20) antibiotics was evaluated and the presence of methicillin resistant Staphylococcus aureus (MRSA) was also determined.

S. aureus was characterized using standard microbiological methods and confirmation was done using the API Staph Identification System. Antibiotic sensitivity of isolates was evaluated by means of agar diffusion technique while the minimum inhibitory concentration (MIC) was established using broth dilution technique for oxacillin, E-test for tetracyclin, and chloramphenicol respectively.

Results: 55.26% of analyzed samples were contaminated with S. aureus. 100% of Methicillin Sensitive S. aureus were resistant to nalidixic acid, 70% to bacitracin, 65% to spiramycin, and 45% to penicillin and fosfomycin. There was no resistance to vancomycin, chloramphenicol, gentamicin and pristamycin among isolates.

A total of 76% of the isolated strains were found to be resistant to at least 4 antibiotics.

One Methicillin Resistant S. aureus strain (4.76%) was detected and showed multiple drug resistance. This resistance was crossed with all beta lactams and its resistance profile to macrolides was constitutive (MLSB const) while aminoglycosides phenotype was ANT (4’) (4’’).

Conclusion: A high prevalence of S. aureus with multiple drug resistance was established. Improved food safety measures are thus necessary to prevent transmission and spread of antimicrobial resistance by these pathogens.

Open Access Original Research Article

Susceptibility of Stored Cocoa Bean Seeds to Ochratoxigenic Fungi in Ondo State, Nigeria

Olukayode O. Orole

Microbiology Research Journal International, Page 1-11
DOI: 10.9734/BMRJ/2016/24137

Aims: To determine the susceptibility of stored cocoa beans to ochratoxigenic fungi in Ondo State, Nigeria.

Place and Duration of Study: The study was carried out at Adekunle Ajasin University, Akungba-Akoko, Ondo State, Nigeria; and cocoa beans samples were collected from stalls in Ondo, Idanre, Oda-Akure, Oba-Akoko, Ile-Oluji, and Ikpenmen towns in Ondo State for a period of twelve months.

Methodology: The serial dilution method was used in plating and isolating the fungi in cocoa bean samples while identification was done conventionally. TLC was used to determine ochratoxin A production by toxin producers.

Results: Cocoa beans collected from stalls in Oba-Akoko had the highest mean fungal incidence of 195 x 105 cfu/g of cocoa bean while those from Idanre stalls had the lowest mean with 4.964 x 105 cfu/g of cocoa bean. Cocoa samples had high fungal incidence between July and September, 2014 compared to other months the study took. Penicillium, Fusarium, Verticicladium, Neocosmospora, Mucor, Rhizopus, Beaveria, Alternaria, Sachharomyces, Phoma were the genera isolated using Czapek Yeast Extract, and TLC screening showed that A. niger, A. ochraceous, A. carbonarius, A. terreus and A. niger are OTA producers. HPLC quantification showed that cocoa bean seeds collected from stalls in Idanre were the least contaminated by OTA while high concentration level of 32.09±22.9 ng/g was detected in Oba-Akoko area of Ondo State.

Conclusion: The study showed that cocoa beans collected from different stalls in Ondo State were susceptible to ochratoxin A contamination, and generally high concentration of the toxin is produced in the wet season than in the dry season. The study further showed that ochratoxigenic moulds infesting stored farm produce such as cocoa bean seeds in Ondo State are majorly the Aspergillus species.

Open Access Original Research Article

Association between Helicobacter pylori Infection and Telomere Length: Effect of Eradication Therapy

Abeer A. Aboelazm, Reem R. Abd El-Glil, Maha Z. Omar

Microbiology Research Journal International, Page 1-9
DOI: 10.9734/BMRJ/2016/23995

Aims: This study aimed to assess the relative telomere lengths (TLs) in gastric mucosa of patients infected with Helicobacter pylori (H. pylori) compared to H. pylori-negative controls and determine the effect of H. pylori eradication therapy on TL.

Place and Duration of Study: This is a seven months case- control study conducted in Hepatology, Gastroenterology and Infectious Diseases and Medical Microbiology & Immunology Departments, Benha University, Egypt.                

Methodology: Relative TLs in gastric mucosa were analyzed by Real Time-Polymerase Chain Reaction (RT-PCR) in 15 H. pylori -positive patients (Group I: 10 patients with gastric ulcer, Group II: 5 patient without gastric ulcer) and 10 H. pylori -negative controls (Group III). Relative TLs were re-evaluated in H. pylori-positive patients 4 weeks after H. pylori eradication therapy.

Results: Highly significant shortening (P<0.001) was observed in TLs in gastric mucosa of H. pylori – positive patients compared to H. pylori – negative controls. Highly significant elongation (P<0.001) was observed after H. pylori eradication therapy. This elongation was significant in both group I and II (P <0.001, 0.01).

Conclusion: H. pylori -positive patients had significantly shorter TLs than H. pylori negative controls. TLs were increased after H. pylori eradication therapy in all patients either with or without gastric ulceration and could be considered as one of preventable methods for gastric cancer.

Open Access Original Research Article

Hospital Antibiogram: A Necessity in Monitoring Sensitivity of Isolates and Rationale Use of Antibiotics’

Ishtiaq Ahmend, Humaira Zafar, Noor Khan Lakhnana, Sundas Ishtiaq, Kiran Tauseef, Mudassira Zahid, Ahsan Kazmi

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/BMRJ/2016/23795

Background: The laid protocols by CDC (center for disease control) narrated the dire need of local anti-biograms. Therefore the current study had been planned to acquire the knowledge about sensitivity pattern of various isolates in different specimens.

Objectives: To identify the local antibiotic sensitivity data against various isolates from different specimens.

Materials and Methods: A descriptive cross sectional study was conducted at the Pathology department of Al Nafees Medical College & Hospital, Islamabad, Pakistan. The duration of study was 04 months i.e 01st June to 01st Oct. 2015. Frequencies and percentages were the numerical variables extracted by using the SPSS version 16.

Results: A total of 336 different specimens were received during study period. E. coli is commonest organism isolated from urine, high vaginal swabs (HVS) and Pus i.e. 60.60%, 100% and 25% respectively. Klebsiella species is the second most common organism (12.12%) followed by Pseudomonas (9.09%) isolated from urine. E. coli, Klebsiella, Pseudomonas and Staphylococcus aureus were isolated at same frequency (25%) from pus. Salmonella typhi was isolated from blood (100%).

The drugs of choice for E. coli are quinolones and aminoglycosides by showing the sensitivity of about 75% each. For Klebsiella pneumoniae, the ideal antibiotics are aminoglycosides (85.1%) and 2nd generation cephalosporins (85%). For Pseudomonas aeruginosa, extended spectrum penicillin, quinolones, 2nd and 3rd generation cephalosporins are the most suitable ones showing the sensitivity of 100% each.

Conclusion: Quinolones, aminoglycosides, 2nd and 3rd generation cephalosporins are the drugs of choices for the treatment of many gram positive and gram negative infections.

Open Access Original Research Article

Comparative Study between Polymerase Chain Reaction and Conventional Methods Used for Diagnosis of Clinically Suspected Onychomycosis

M. A. Abd El-Wahab, W. S. El Naghy, R. A. El Tatawy

Microbiology Research Journal International, Page 1-9
DOI: 10.9734/BMRJ/2016/24852

Background: Proper treatment of onychomycosis depends mainly on accurate diagnosis. Diagnosis using conventional methods (direct microscopy and culture) is still insensitive and time consuming so we aimed to evaluate the commercially available PCR as a rapid method used for detection of fungal elements especially Trichophyton rubrum in nail specimens with clinically suspected onychomycosis and compare the results with conventional diagnostic methods.

Methodology: This study included 50 patients with clinically suspected onychomycosis of nails. Nail specimens were obtained by scraping of the diseased nail(s) with a sterile scalpel blade and collected in a sterile container. Each specimen was divided into 3 portions and processed immediately or kept at room temperature until use. The first part was examined by direct microscopy with 20% KOH, the second part was cultured on Sabouraud dextrose agar (SDA) and the third part was subjected to pan-fungal PCR for detection of fungal infection followed by specific PCR for identification of T. rubrum.

Results: Regarding all fungal isolates KOH microscopy, culture and pan-fungal PCR respectively yielded positive rates of 27 (54%), 26 (52%) and 37 (74%). Pan-fungal PCR picked up 11 specimens missed by culture. Regarding T. rubrum, KOH microscopy, culture and T. rubrum specific PCR respectively yielded positive rates of 3 (6%), 11 (22%) and 20 (40%). T. rubrum specific PCR picked up 9 specimens missed by culture. Of the 26 specimens that were culture positive, 14 grew dermatophytes, 9 Candida spp and 3 non-dermatophytic molds. These results emphasize the superiority of PCR over conventional methods.

Conclusion: This study demonstrates that both pan-fungal and T. rubrum specific PCR have higher positive rates for detection of fungal infections of the nails compared with KOH microscopy or culture. We suggest that PCR should be used as a complementary method for confirmation of clinically suspected onychomycosis.

Open Access Original Research Article

Food-borne Bacteria: Occurrences, Multidrug Resistant Patterns and Susceptibility to Aqueous Leaf Extracts of Acalypha hispida (Linn)

O. J. Akinjogunla, K. O. Adewumi, M. U. Okon

Microbiology Research Journal International, Page 1-11
DOI: 10.9734/BMRJ/2016/24381

The occurrence and susceptibility of multidrug resistant (MDR) bacteria obtained from food to aqueous leaf extracts of Acalypha hispida (ALEAH) were determined using standard bacteriological and disc diffusion methods. In rice, Total Heterotrophic Counts (THC), Total Coliform Counts (TCC) and Total Feacal Counts (TFC) ranged from 3.3x103 to 1.9x104 (CFU/g), 1.9x103 to 3.6x103 (CFU/g) and 1.0x102 to 1.6x102 (CFU/g), respectively. In ‘garri’, highest THC (2.0x 104 CFU/g), highest TCC (1.2x 104 CFU/g) were obtained in sample GA-03 and lowest TCC (2.0x 103 CFU/g) were obtained in samples GA-04 and GA-11. In ‘fufu’, the THC and TCC ranged from 7.3x 103 to 9.5x 103 (CFU/g) and 2.6x 103 to 4.6x 103 (CFU/g), respectively, while ≥ 70% ‘fufu’ had no TFC. Eight genera (Escherichia, Staphylococcus, Enterobacter, Streptococcus, Klebsiella, Proteus, Pseudomonas and Salmonella were obtained in the food samples. Growths of single bacterial isolate were obtained in 41.7% of rice, while co-contamination with two and three isolates were observed in 33.3% and 25.0% samples, respectively. Only 50.0%, 16.7% and 33.3% of ‘garri’ had growth of single, two and three bacterial isolates, respectively, while between 1 (8.3%) to 5 (41.7%) of ‘fufu’ and rice had growth of two and three bacterial isolates. The ALEAH contained alkaloids, cardiac glycoside, phenolics, saponin, tannin, flavonoids, deoxy- sugar, anthraquinones and phlobatanins. The results showed that between 45.5% to 66.7% bacterial isolates were multidrug resistant. Of 56.3% MDR S. aureus obtained from the boiled rice, fufu’ and ‘garri’ only SAF4 and SAR3 had the same resistance patterns. The discs containing 20 mg/ml and 80 mg/ml ALEAH showed the lowest and highest activity of 6.7±1.8 mm and 13.6±1.0 mm, respectively. Streptococcus spp SPF1; E. coli ECG3, and P. aeruginosa PAF2 were resistant to 20 mg/ml and 40 mg/ml of the ALEAH. Conclusively, there is need to  always evaluate the bacteriological quality of the food sold in the fast food centres / restaurants from time to time and train the food handlers on food safety practices.