Open Access Original Research Article

Incidence of Enterobacteriaceae in Retail Meat Samples and Their Antibiotic Susceptibility

Shreyasi Dubey, Pinki Saini, Divyanshi Singh, Priyanka Singh

Microbiology Research Journal International, Page 1-9
DOI: 10.9734/MRJI/2018/40767

The study aims at accounting the prevalence of pathogenic bacteria within the family Enterobacteriaceae in retail meat samples. Eighty samples were collected from four different locations of Allahabad region. Isolation was done using selective plating according to ISO Standard; ISO 21528-1:2004. Differentiation and characterization of different isolates was based on their growth characteristics on specific culture media, their biochemical confirmatory tests and Gram-staining reactions. Total soluble proteins of the isolates were estimated by Biuret method. Antibiotic susceptibility of the isolates was tested against antibiotics including ampicillin, streptomycin and ciprofloxacin at different concentrations. A total of 62 isolates were obtained and identified as Klebsiella planticolaCitrobacter youngaeEnterobacter sp, E. cloacaeK. ornithinolytica and K. pneumonia. All the isolates were susceptible to ampicillin, streptomycin and ciprofloxacin at concentrations of 10, 50, 80 and 100 mg/ml. The virulent proteins were highest in Enterobacter (127.3 mg/ml) followed by C. youngae (119.4 mg/ml). Meat sold in the local markets of Allahabad showed presence of pathogenic bacteria belonging particularly to the family Enterobacteriaceae; indicating poor hygienic conditions as well as improper storage environment. The results revealed that K. pneumonia (29.9%) represented the major part of bacterial flora, in the samples followed by C. youngae(20%).

Open Access Original Research Article

Phenotypic Identification and Detection of BlaCTX and TetM in Klebsiella Species from Urine Samples of Patients in Ile-Ife, Nigeria

Simaren Yetunde Olawale, Oluduro Anthonia Olufunke

Microbiology Research Journal International, Page 1-11
DOI: 10.9734/MRJI/2018/23733

Klebsiella species from urine samples patients in Ile-Ife were investigated. Isolation, characterization and identification of the isolates were done using standard microbiological techniques. Antibiotic sensitivity of the Klebsiella isolates to twelve different antibiotics was determined by Kirby-Bauer’s disc diffusion method on Mueller-Hinton agar plates. Plasmid DNA in representative multiple antibiotic resistant isolates was detected by alkaline lysis (TENS buffer). Resistance (bla CTX, tet M) genes were detected by Polymerase Chain Reaction (PCR). Statistical analysis of data obtained was done using SPSS-17.0. A total of thirty-two Klebsiella sp. (7.02%) were isolated from 456 urine samples of patients; 50 urine samples collected from apparently healthy individuals served as control. Antibiotic resistance was high and also varied among the isolates with a range of 37.5% to 100%. Most of the isolates had multiple antibiotic resistance (MAR) to at least three different classes of antibiotics. There was diversity in the MAR patterns with 21 different antibiotypes which ranged from two to ten different antibiotics. Plasmid DNA of molecular weight of 900 bp to 9416 bp was detected in some representative multiple antibiotic resistant isolates. Three isolates harboured bla CTX (543 bp) gene which codes for resistance to beta-lactams. However, tet M (974 bp) gene which codes for resistance to tetracycline was not detected in any of the isolates. The study concluded that there were multiple antibiotic resistant Klebsiella species from urine samples of patients which harboured plasmid DNA and resistance (bla CTX) gene which is of great health and economic consequences.

Open Access Original Research Article

Molecular Characterization of Non-O157 Shiga Toxin-producing E. coli Detected in Raw Milk and Some Dairy Products

Mona A. El-Zamkan, Karima G. Abdel Hameed

Microbiology Research Journal International, Page 1-14
DOI: 10.9734/MRJI/2018/40434

Aims: The aim of this study was to detect non-O157 Shiga toxin producing E. coli in raw milk and some of its products in Qena, Egypt.

Study Design: An exploring, evaluating study. 

Place and Duration of Study: The study was conducted at Department of Food Hygiene and Control, Faculty of Veterinary Medicine, South Valley University, Qena, Egypt.

Methodology: A total of 90 samples of raw milk, white cheese and small scale ice cream sold in local markets in Qena city, Egypt were collected and investigated for the presence of non-O157 STEC. The tested products were screened for the presence of Shiga toxin by ELISA Kits. A loopful from samples that gave a positive reading in ELISA test was streaked onto SHIBAM plates for isolation of non-O157 STEC. Then the obtained isolates were molecularly characterized and serotyped.

Results: Shiga toxin was detected in 12.2% of the examined samples by using ELISA. From the ELISA-positive samples, 28 non-O157 E. coli strains were isolated and molecularly characterized by the presence of stx1, stx2, eaeA and the genes. All non-O157 STEC obtained from raw milk samples carried stx2 gene only and lacked stx1, eaeA and hly genes. While 94.1% and 5.8% of the non-O157 STEC obtained from white cheese samples harbored stx1 and stx2, respectively, eaeA and hly genes also could be detected in 82.4, and 11.7% of non-O157 STEC strains isolated from white cheese, respectively. One non-O157 STEC isolates could be obtained from the ice cream samples, and it harbored stx1, eaeA and hly genes. 

Conclusion: The presence of the non-O157 STEC in the examined samples reinforces the idea that these products exhibit a potential health hazard to the consumers since the hazard to the consumer is associated with the virulence of the detected strains in the investigated dairy products.

Open Access Original Research Article

Prevalence and Characterization of Staphylococcus aureus Isolated from Bulk Tank Milk Dairy Cow Farms in West Bank-Palestine

Ghaleb Adwan, Haya Isayed

Microbiology Research Journal International, Page 1-13
DOI: 10.9734/MRJI/2018/40074

Aims: The current study aimed to characterize and document the occurrence of Methicillin-sensitive S. aureus (MSSA) and Methicillin-resistant S. aureus (MRSA) in bulk tank milk (BTM) samples, farm workers and the environmental surfaces from bovine dairy farms, antibiotic resistance rate, and genetic characterization of clones for both MSSA and MRSA using Enterobacterial Repetitive Intergenic (ERIC-PCR) and Staphylococcal protein A (spA) gene typing.

Methodology: A total of 57 bovine BTM samples, 45 samples from farm environmental surfaces and 16 nasal swab samples from farm workers, were collected from 12 bovine dairy farms in the Jenin district in northern West Bank-Palestine, between September and October, 2017.

Results: Results of this research showed that 83.3% of farms had at least one BTM sample contaminated with Staphylococcus aureus. Also, 75% and 58.3% of farms had contaminated environmental surfaces and farm worker carriers with S. aureus, respectively. Also, it showed that 58.3% of farms had at least one BTM sample contaminated with MRSA, while 50% of farms had MRSA contaminated environmental surfaces and MRSA farm worker carriers. Results of the current study also showed that 68.4% and 45.6% of bovine BTM samples were contaminated with S. aureus and MRSA, respectively. S. aureus recovered from different sources showed a high level of resistance to many different antibiotics. Also, results of this study showed that 100% and 69.2% of MRSA and MSSA strains, respectively, isolated from BTM samples were multi-drug resistant (MDR). ERIC-PCR profile and spa typing showed that some strains of the same clone had been isolated from diverse sources from different farms. This evidence suggested that these strains of the same clone or spa type could be circulated between cattle, environment, and humans.

Conclusion: Results of this study showed that Palestine BTM samples are a common source of MRSA. The presence of MRSA isolates in BTM may present a potential public health risk. Therefore, careful monitoring of the resistance status of S. aureus in cattle, farm workers, and dairy environmental surfaces is required due to some clones that circulate among them and play a major threat to farm workers who are in close contact with cattle.

Open Access Original Research Article

Undiagnosed Chronic Hepatitis B Infection and HIV Type 1 Drug Resistance Profile in AIDS Patients Receiving Tenofovir-containing Antiretroviral Regimens: Considerations for Monitoring Resistant HIV Variants during Treatment

Charles Kouanfack, Mathurin Kowo, Emmanuel Sako Haddison, Veronica Matehbi Aletum, Desire Takou, Patrick Awoumou, Serge Christian Tchokonte, Samuel Sosso, Njoya Oudou, Judith Ndongo Torimiro

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/MRJI/2018/40801

Background: During antiretroviral therapy (ART), resistant HIV-1 variants may be selected resulting in clinical failure. Co-infection rate of Hepatitis B virus in AIDS patients in Cameroon is 11.8%, but few studies have described the profile of resistance associated mutations (RAMs) in HIV in these patients on first-line ART containing antiviral drugs against hepatitis B infection. Thus, we aimed to determine the rate of HBV infection and profile of HIV-1 RAMs in AIDS patients on ART.

Methods:  A cross-sectional study was carried out from November 2013 to April 2014 in two AIDS Treatment Centres in Yaoundé, Cameroon. Ninety-six adult AIDS patients on tenofovir-containing regimens were tested for HBsAg by serology. Direct sequencing of amplicons generated for the HIV-1 protease/reverse transcriptase region, was performed for 21 HIV/HBV co-infected patients. The Stanford HIV Drug Resistance Database tool was used to predict RAMs, and genotyping was determined by phylogeny. 

Results: Overall, 21 patients were co-infected with HBV (21.9%). Eighteen (85.7%) of these were infected with recombinant variants, and HIV-1 CRF02_AG was most frequently identified (52.9%), and 15 (71.4%) harboured at least one RAM. Prediction of resistance to NNRTIs was reported in 14 (66.7%), and among 13 (61.9%) to NRTIs, 2 (9.5%) to PIs, and 8 (38.1%) carried Thymidine Analog Mutations (TAMs) of M184VI (61.9%), V75I (23.8%), T215F (23.8%), M41L (19.0%) and K70R (19.0%).

Conclusion: Rate of HBV infection and frequency of HIV-1 RAMs among AIDS patients on ART is high. The observed NNRTI RAMs may affect the susceptibility of efavirenz. Thus, the need to monitor HIV-1 drug resistance profile during treatment with unsuppressed viral load.