scholarly journals MOLECULAR DETECTION AND CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS ISOLATED FROM RAW MILK SOLD IN DIFFERENT MARKETS OF BANGLADESH

2017 ◽  
Vol 14 (2) ◽  
pp. 277-282 ◽  
Author(s):  
M. A. Islam ◽  
S. M. L. Kabir ◽  
M. T. Rahman
Keyword(s):  
Molecular Detection ◽  
Culture Media ◽  
Raw Milk ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Selective Media ◽  
Milk Samples ◽  
Antimicrobial Sensitivity ◽  
Antimicrobial Sensitivity Test

The study was intended for molecular detection of S. aureus isolated from raw cow’s milk. A total of 20 milk samples were collected from different upazila markets of Jamalpur, Tangail, Kishoreganj and Netrokona districts of Bangladesh. Milk samples were cultured onto various culture media for the isolation of bacteria. The isolated bacteria were identified by studying cultural properties on different selective media, biochemical tests, and finally by PCR. Out of 20 samples, 15 (75%) milk samples were found to be positive for S. aureus. S. aureus specific 16S rRNA gene was amplified from all isolates and identified as S. aureus. Antimicrobial sensitivity test was carried out to ascertain the susceptibility of the organism to various antibiotics. Its results showed that the S. aureus isolates were resistant to amoxicillin (100%), erythromycin (73.33%) and tetracycline (73.33%) but sensitive to azithromycin (93.33%), ciprofloxacin (93.33%), gentamicin (100%), norfloxacin (86.67%) and streptomycin (86.67%).

scholarly journals MOLECULAR DETECTION AND CHARACTERIZATION OF ESCHERICHIA COLI ISOLATED FROM RAW MILK SOLD IN DIFFERENT MARKETS OF BANGLADESH

2017 ◽  
Vol 14 (2) ◽  
pp. 271-275 ◽  
Author(s):  
M. A. Islam ◽  
S. M. L. Kabir ◽  
S. K. Seel
Keyword(s):  
Escherichia Coli ◽  
Molecular Detection ◽  
Culture Media ◽  
Raw Milk ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
E Coli ◽  
Milk Samples ◽  
Antimicrobial Sensitivity

     The study was intended for molecular detection of E. coli isolated from raw cow’s milk. A total of 20 milk samples were collected from different upazila markets of Jamalpur, Tangail, Kishoreganj and Netrokona districts of Bangladesh. Milk samples were cultured onto various culture media for the isolation of bacteria. The isolated bacteria were identified by studying staining characteristics, cultural properties on different selective media, biochemical tests, catalase and coagulase test, and finally by PCR. Out of 20 samples, 15 (75%) milk samples were found positive for E. coli. 15 Escherichia coli isolates were amplified by 16S rRNA gene based PCR. Antimicrobial sensitivity test was carried out to ascertain the susceptibility of the organism to various antibiotics. Its results showed that the E. coli isolates were resistant to amoxycillin (86.67%) and erythromycin (73.33%) but sensitive to azithromycin (53.33%), ciprofloxacin (86.67%), gentamicin (86.67%), norfloxacin (80%) and streptomycin (66.67%).

scholarly journals Molecular Detection and Antimicrobial Susceptibility of Salmonella Species in Chickens

2021 ◽  
Vol 10 (9) ◽  
pp. 560-566
Author(s):  
Gloria Sugun Ehizibolo David ◽  
Ebenezer Bukola Ogundejii Kaikabo Ahmed ◽  
Agada O. Godwin Benshak Audu ◽  
Sugun Manasa
Keyword(s):  
Antimicrobial Susceptibility ◽  
Biochemical Analysis ◽  
Molecular Detection ◽  
Biochemical Tests ◽  
Salmonella Spp ◽  
Selective Media ◽  
Antimicrobial Sensitivity ◽  
Cultural Properties ◽  
Plateau State ◽  
Antimicrobial Sensitivity Test

The study was intended for molecular detection of Salmonella Spp isolated from chicken. A total of 160 samples comprising of 40 liver, 40 spleen, 40 lungs and 40 intestines were collected directly into buffered peptone in universal bottles at the poultry slaughter houses of the four districts in Jos South Local Government area of Plateau State. The samples were enriched in 10 ml of Rappaport-Vassiliades broth and cultured onto Xylems Lysine Deoxycholate (XLD) agar for the isolation of bacteria. The isolated bacteria were identified by studying staining characteristics, cultural properties on different selective media, biochemical tests, catalase and coagulase test, and finally by PCR. Out of 160 samples, 65 (41%) samples were found positive for Salmonella Spp on XLD and 24 (37%) positive by biochemical analysis. Two(2) Salmonella isolates were amplified by 942 bp gene based PCR. Antimicrobial sensitivity test was carried out to ascertain the susceptibility of the organism to various antibiotics. Its results showed that the Salmonella isolates were resistant to amoxycillin (100%) and erythromycin (100%), gentamicin (100%), Clindamycin (100%) streptomycin (100%), tetracycline (100%), sulphamethoxazole/trimethoprim (100%) but sensitive to Ceftiofur (100%).

scholarly journals Microbial assessment of milk collected from different markets of Mymensingh, Gazipur and Sherpur districts of Bangladesh and determination of antimicrobial resistance patterns of the isolated bacteria

2017 ◽  
Vol 1 (1) ◽  
pp. 7-16
Author(s):  
Mohammad Farhad Hossain ◽  
Md Tanvir Rahman ◽  
SM Lutful Kabir
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Bacterial Load ◽  
Microbiological Quality ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Selective Media ◽  
E Coli ◽  
Milk Samples ◽  
Unpasteurized Milk

The study was conducted to determine the total viable count (TVC) and total coliform count (TCC) of unpasteurized, pasteurized and UHT milk samples to assess their microbiological quality. A total of 60 milk samples were collected from different markets of Mymensingh, Gazipur and Sherpur districts. The TVC of milk was performed to determine the bacterial load in supplied milk samples and TCC to determine the coliform bacterial load in collected milk samples. Milk samples were cultured onto various selective media for the isolation of bacteria. The isolated bacteria were identified by cultural properties on different selective media, biochemical tests and finally by PCR. Out of 60 samples 20 unpasteurized milk samples from different markets were found positive for Staphylococcus aureus and 19 unpasteurized milk samples were found positive for Escherichia coli. All the S. aureus were found positive for S. aureus specific 16S rRNA gene by PCR. Out of 19 isolates of E. coli, 15 were found positive for E. coli 16S rRNA gene by PCR. Results of antimicrobial susceptibility test showed that most of the isolates of S. aureus and E .coli were susceptible to azithromycin, streptomycin, gentamicin, norfloxacin, tetracycline and ciprofloxacin but resistant to amoxicillin and erythromycin. The findings of this study revealed the presence of multidrug resistant S. aureus and E. coli in unpasteurized milk samples that posseses a serious threat to public health. Asian Australas. J. Food Saf. Secur. 2017, 1 (1), 7-16

scholarly journals Isolation and Molecular Characterization of Shigatoxigenic O157 and Non-O157 Escherichia coli in Raw Milk Marketed in Chittagong, Bangladesh

2021 ◽  
Author(s):  
Md. Kauser-Ul Alam ◽  
Nazmul Sarwar ◽  
Shireen Akther ◽  
Monsur Ahmad ◽  
Paritosh Kumar Biswas
Keyword(s):  
Escherichia Coli ◽  
Molecular Characterization ◽  
Raw Milk ◽  
Selective Medium ◽  
Enterohemorrhagic Escherichia Coli ◽  
Macconkey Agar ◽  
Biochemical Tests ◽  
E Coli ◽  
Milk Samples

Background: Quality and microbial safety of milk is demanding day by day as it is considered as a host for pathogenic and spoilage microorganisms. In this study, isolation and molecular characterization of shigatoxigenic O157 and non-O157 Escherichia coli in raw milk marketed in Chittagong, Bangladesh were done on 186 raw milk samples in Bangladesh. Methods: MacConkey agar was initially used to screen for the presence of E. coli and the suspected growth as evidenced by large pink colonies on MacConkey agar. Finally the organism was verified by plating through Eosin Methylene Blue (EMB) agar (a selective medium for E. coli where it produces metallic sheen) and applying standard biochemical tests for E. coli. The presence of virulent genes, Shiga-like toxin (stx1 and stx2), Intimin (eaeA), O157 antigen rfbE and Enterohemorrhagic Escherichia coli (EHEC) Hemolysin (EHEC) hlyA in the contaminating E. coli population was determined by polymerase chain reaction (PCR) run on a thermocycler (Applied Biosystem, 2720 thermal cycler, Singapore). Result: Among the raw milk samples, 33 samples were identified as E.coli positive and among the isolates, 6 (18.18%) were identified as possible EHEC O157 and rest of the isolates (81.82%) were considered as probable non EHEC O157. About, 3.23% (186 samples) EHEC O157 was isolated from raw milk samples. Then all the 33 isolates were taken under PCR assay for the identification of five virulent genes Stx1, Stx2, eaeA, rfbE and hlyA. No virulent genes were found in non- EHEC O157 isolates, but 4 stx2 (66.67%) and 1 hlyA (16.67%) gene were observed in another 4 EHEC O157 isolates out of 6, but one isolates contained the both genes and hence the prevalence of STEC was 2.15% in raw milk. Result indicated poor hygienic standard of raw milk from uncontrolled environments and the increased public health risk of those consuming raw milk from such uncontrolled sources.

Phylogeny Characterization of Seb Gene Encoding Enterotoxins in Staphylococcus aureus Isolated from Raw Milk and Cheese

2019 ◽  
Vol 9 (o3) ◽  
Author(s):  
Fatima N. Aziz ◽  
Laith Abdul Hassan Mohammed-Jawad
Keyword(s):  
Staphylococcus Aureus ◽  
Food Poisoning ◽  
Raw Milk ◽  
Staphylococcal Enterotoxin B ◽  
Human Pathogen ◽  
Conventional Pcr ◽  
Biochemical Tests ◽  
Specific Primers ◽  
Gene Encoding

Food poisoning due to the bacteria is a big global problem in economically and human's health. This problem refers to an illness which is due to infection or the toxin exists in nature and the food that use. Milk is considered a nutritious food because it contains proteins and vitamins. The aim of this study is to detect and phylogeny characterization of staphylococcal enterotoxin B gene (Seb). A total of 200 milk and cheese samples were screened. One hundred ten isolates of Staphylococcus aureus pre-confirmed using selective and differential media with biochemical tests. Genomic DNA was extracted from the isolates and the SEB gene detects using conventional PCR with specific primers. Three staphylococcus aureus isolates were found to be positive for Seb gene using PCR and confirmed by sequencing. Sequence homology showed variety range of identity starting from (100% to 38%). Phylogenetic tree analyses show that samples (6 and 5) are correlated with S. epidermidis. This study discovered that isolates (A6-RLQ and A5-RLQ) are significantly clustered in a group with non- human pathogen Staphylococcus agnetis.

scholarly journals Taxonomic characterization of nine strains isolated from clinical and environmental specimens, and proposal of Corynebacterium tuberculostearicum sp. nov.

2004 ◽  
Vol 54 (4) ◽  
pp. 1055-1061 ◽  
Author(s):  
Carole Feurer ◽  
Dominique Clermont ◽  
François Bimet ◽  
Adina Candréa ◽  
Mary Jackson ◽  
...  
Keyword(s):  
Gene Sequence ◽  
Fatty Acid Content ◽  
Food Samples ◽  
Cell Protein ◽  
Rrna Gene ◽  
Biochemical Tests ◽  
Taxonomic Analysis ◽  
Gene Sequence Analysis ◽  
Taxonomic Characterization

Nine unidentified Gram-positive, lipophilic corynebacteria were isolated from clinical and food samples and subjected to a polyphasic taxonomic analysis. The bacteria were distinguished from Corynebacterium species with validly published names by biochemical tests, fatty acid content and whole-cell protein analysis. Comparative 16S rRNA gene sequence analysis demonstrated unambiguously that the nine strains were related phylogenetically to the species ‘Corynebacterium tuberculostearicum’ and represented a distinct subline within the genus Corynebacterium. On the basis of both phenotypic and phylogenetic evidence, the formal description of Corynebacterium tuberculostearicum sp. nov. is proposed. The type strain of C. tuberculostearicum is Medalle XT (=LDC-20T=CIP 107291T=CCUG 45418T=ATCC 35529T).

scholarly journals Isolation and molecular characterization of Mycobacterium tuberculosis complex isolated from raw milk in some dairy farms in Egypt

2016 ◽  
Vol 5 (2) ◽  
pp. 105
Author(s):  
Heba Hussien ◽  
Eman Mahrous
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Culture Method ◽  
Raw Milk ◽  
Accurate Method ◽  
Tuberculosis Complex ◽  
Milk Samples ◽  
Source Of Infection

<p>This study was conducted to detect <em>Mycobacterium tuberculosis</em> complex in milk in three Egyptian Governorates; El-Sharkia, El-Menoufia and El-Behera Governorates. 300 milk samples were collected from tuberculin positive cases, 18 (6.0%) were shedding <em>Mycobacterium tuberculosis</em> complex in their milk which detected by real time PCR. On another hand, 170 milk samples were collected from tuberculin negative cases, 5 (2.9%) were shedding <em>Mycobacterium tuberculosis</em> complex in their milk which detected by real time PCR. All milk samples were examined by three techniques including Microscopic examination, culture and real time PCR. Real time PCR is more rapid and accurate method than microscopic and culture method. The isolated colonies from culture were examined by Multiplex PCR to demonstrate the source of infection either human or animal source.</p>

scholarly journals Molecular Detection and Antibiotic Sensitivity of Salmonella Species Isolated from Goat Feces in Sylhet District of Bangladesh

2021 ◽  
Vol 11 (3) ◽  
pp. 395-401
Author(s):  
Md. Abdus Sabur ◽  
Mouri Rani Das ◽  
Md Bashir Uddin ◽  
Md. Mahfujur Rahman ◽  
Md. Rafiqu Islam ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Antibiotic Sensitivity ◽  
Culture Methods ◽  
Specific Primers ◽  
Antimicrobial Sensitivity ◽  
Conventional Culture ◽  
Antibiotic Sensitivity Test ◽  
One Year ◽  
Pcr Techniques

The present study aimed at the molecular detection of Salmonella species from feces of goats and the characterization of the isolated Salmonella by biochemical and antimicrobial sensitivity techniques. A total of 220 goat feces samples were collected, of which 27 (12.27%) were positive for Salmonella by conventional culture methods and 20 (9.09%) by biochemical and PCR techniques. The prevalence was higher in goats under one year of age (20%), compared to older animals aged one to two years (7.8%) and more than two years of age (4.7%), respectively. Moreover, the prevalence of diarrheic goats was significantly higher (38.46%) than healthy animals (2.76%). DNA was extracted from Salmonella strains and amplified by PCR using the specific primers of Salmonella invasion gene (invA gene). The antibiotic sensitivity test indicated that Ciprofloxacin (100 percent sensitivity), Gentamycin (100 percent sensitivity), and Neomycin (100 percent sensitivity) were the most effective antibiotics for the majority of Salmonella isolates. On the other hand, Salmonella isolates were found to have substantially high resistance to Erythromycin (100%), Amoxicillin (100%), Trimethoprim-Sulfamethoxazole (81.48%), Streptomycin (62.96%), and Tetracycline (55.56 percent). Since the rate of Salmonella carriers was relatively high, eating goat meat could increase the risk of foodborne salmonellosis.

Occurrence of Listeria monocytogenes in Raw Milk in Nebraska

1988 ◽  
Vol 51 (11) ◽  
pp. 840-841 ◽  
Author(s):  
MICHAEL B. LIEWEN ◽  
MARK W. PLAUTZ
Keyword(s):  
Listeria Monocytogenes ◽  
Dairy Farms ◽  
Raw Milk ◽  
Storage Tanks ◽  
Biochemical Tests ◽  
Bulk Storage ◽  
Milk Samples ◽  
Listeria Species ◽  
Enrichment Procedure

Raw milk samples were obtained from bulk storage tanks of individual dairy farms in eastern Nebraska during February and July of 1986. One hundred different farms were tested during each period. One-tenth ml of each sample was plated directly onto McBride's Listeria Agar (MLA) and 30 ml was subjected to a four-week cold enrichment procedure. Suspect colonies from MLA were subjected to biochemical tests to confirm identity. Nine percent of all raw milk samples examined were determined to be positive for Listeria species after the cold enrichment procedure. Four percent contained L. monocytogenes and five percent contained L. innocua. Six percent and two percent of samples were found to contain L. monocytogenes in February and July respectively.

scholarly journals Screening and enumeration of lactic acid bacteria in milk using three different culture media in Petrifilm™ Aerobic Count plates and conventional pour plate methodology

2007 ◽  
Vol 74 (4) ◽  
pp. 387-391 ◽  
Author(s):  
Maria BT Ortolani ◽  
Gabriela N Viçosa ◽  
Vanerli Beloti ◽  
Luís A Nero
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Culture Media ◽  
Skim Milk ◽  
Raw Milk ◽  
Excellent Correlation ◽  
Anaerobic Conditions ◽  
Milk Samples ◽  
Experimental Inoculation

This study aimed to compare Petrifilm™ Aerobic Count (AC) plates and the conventional pour plate methodology using de Mann-Rogosa-Sharpe (MRS), Kang-Fung (KF) and Kang-Fung-Sol (KFS) culture media for screening and enumeration of lactic acid bacteria (LAB) in milk. Suspensions of 10 LAB species in reconstituted powder skim milk and 30 raw milk samples, without experimental inoculation, were tested. For selective enumeration, all samples were previously diluted in MRS, KF and KFS broths and then plated in Petrifilm™ AC and conventional pour plate methodology, using the same culture media with added agar. All plates were incubated at 37°C for 48 h in anaerobic conditions. Differences in the counts were observed only for raw milk samples using KFS in conventional methodology, when compared with the counts obtained from MRS and KF (P⩽0·05). The results showed excellent correlation indexes between both methodologies using the three culture media for LAB suspensions (r=0·97 for MRS, KF and KFS). For raw milk samples, the correlation indexes were excellent (r=0·97, for MRS) and good (r=0·84 for KF, and r=0·82 for KFS), showing some interference in Petrifilm™ AC when supplements were added, especially lactic acid. These results indicate the possibility of using Petrifilm™ AC plates for enumeration of LAB in milk, even with the use of selective supplements.

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