Occurrence and Characterization of Extended Spectrum β- lactamase Producing E. coli and Salmonella spp. from Raw Milk Samples in  Wayanad District, Kerala, India

The study was intended for molecular detection of E. coli isolated from raw cows 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%).

Download Full-text

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

Asian Journal of Dairy and Food Research ◽

10.18805/ajdfr.dr-178 ◽

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.

Download Full-text

Prevalence and Characterization of Pathogenic Bacteria in Bulk Tank Raw Milk, Thailand

Walailak Journal of Science and Technology (WJST) ◽

10.48048/wjst.2020.4177 ◽

2020 ◽

Vol 17 (6) ◽

pp. 588-599

Author(s):

Chanida KUPRADIT ◽

Sasidhorn INNOK ◽

Jirayus WORARATPHOKA ◽

Mariena KETUDAT-CAIRNS

Keyword(s):

Foodborne Pathogens ◽

Pathogenic Bacteria ◽

Raw Milk ◽

Salmonella Spp ◽

Milk Samples ◽

Isolation And Characterization ◽

Lack Of Information ◽

Enterotoxin Genes ◽

Bulk Tank

Although there are a number of raw milk collection centers in Nakhon Ratchasima, there is a lack of information with regard to the process of isolation and characterization of foodborne pathogens in raw milk. Therefore, the purpose of this research was to investigate the prevalence and characterization of foodborne pathogens, including Bacillus cereus, Escherichia coli, Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus from 33 raw milk samples from 9 different raw milk collection centers located in 8 districts in Nakhon Ratchasima, Thailand. This study was conducted from January to March 2016. Results revealed that the contaminations of L. monocytogenes and Salmonella spp. were not detected in any of the raw milk samples tested. The prevalence of B. cereus, E. coli, and S. aureus in raw milk samples was found to be 9 % (10 - 2.0 ×104 CFU/ml), 42.4 and 54 % (85 - 2.7 ×104 CFU/ml), respectively. The distribution of virulence genes was tested in B. cereus and S. aureus using gene specific primers by polymerase chain reaction. Out of the 29 analyzed coagulase-positive S. aureus isolates, 27 isolates (93 %) were positive for eap gene amplification and 14 isolates (48 %) showed amplicon of eap gene and all 5 enterotoxin genes, including seG, seGV, seI, seIV, and seM genes. All 8 B. cereus isolates tested showed positive PCR result with enterotoxin FM (entFM) gene but they showed negative with hemolysin gene (hblA and hblD genes) amplifications. It was inferred from these findings that bulk tank milk is a potential source of S. aureus and B. cereus in milk.

Download Full-text

Prevalence of Mastitis and Antibiotic Resistance of Bacterial Isolates from CMT Positive Milk Samples Obtained from Dairy Cows, Camels, and Goats in Two Pastoral Districts in Southern Ethiopia

Animals ◽

10.3390/ani11061530 ◽

2021 ◽

Vol 11 (6) ◽

pp. 1530

Author(s):

Amanuel Balemi ◽

Balako Gumi ◽

Kebede Amenu ◽

Sisay Girma ◽

Muuz Gebru ◽

...

Keyword(s):

Antibiotic Resistance ◽

Dairy Cows ◽

Raw Milk ◽

Multidrug Resistant ◽

Southern Ethiopia ◽

Resistant Bacteria ◽

Coagulase Negative Staphylococcus ◽

Bacterial Isolates ◽

E Coli ◽

Milk Samples

A study was carried out from August 2017 to February 2018 on lactating dairy cows, one-humped dromedary camels, and goats to determine mastitis in the Bule Hora and Dugda Dawa districts of in Southern Ethiopia. Milk samples from 564 udder quarters and udder halves from 171 animals consisting of 60 dairy cows, 51 camels, and 60 goats were tested for mastitis. Sixty-four positive udder milk samples were cultured, and bacterial mastitis pathogens were isolated and identified. The antibiotic resistance of bacterial isolates from milk with mastitis was tested against nine antimicrobials commonly used in the study area. Cow- and quarter-level prevalence of mastitis in dairy cows, camels, and goats was 33.3%, 26.3%, and 25% and 17.6%, 14.5%, and 20%, respectively. In cattle, the prevalence was significantly higher in Dugda Dawa than in Bule Hora. Major bacterial isolates were coagulase-negative Staphylococcus species (39.1%), S. aureus (17.2%), S. hyicus (14.1%), and S. intermedius and Escherichia coli (9.4% each). In camels, udder abnormality and mastitis were significantly higher in late lactation than in early lactation. Mastitis tends to increase with parity in camels. E. coli isolates were highly resistant to spectinomycin, vancomycin, and doxycycline, whereas most S. aureus isolates were multidrug-resistant. Most of the rural and periurban communities in this area consume raw milk, which indicates a high risk of infection with multidrug-resistant bacteria. We recommend a community-focused training program to improve community awareness of the need to boil milk and the risk of raw milk consumption.

Download Full-text

Isolation and Characterization of Bacteriophage ZCSE6 against Salmonella spp.: Phage Application in Milk

Biologics ◽

10.3390/biologics1020010 ◽

2021 ◽

Vol 1 (2) ◽

pp. 164-176

Author(s):

Abdallah S. Abdelsattar ◽

Anan Safwat ◽

Rana Nofal ◽

Amera Elsayed ◽

Salsabil Makky ◽

...

Keyword(s):

Pathogenic Bacteria ◽

Raw Milk ◽

Salmonella Spp ◽

Isolation And Characterization ◽

Food Borne ◽

Wide Range ◽

Almost All ◽

And Control ◽

Milk And Dairy Products

Food safety is very important in the food industry as most pathogenic bacteria can cause food-borne diseases and negatively affect public health. In the milk industry, contamination with Salmonella has always been a challenge, but the risks have dramatically increased as almost all bacteria now show resistance to a wide range of commercial antibiotics. This study aimed to isolate a bacteriophage to be used as a bactericidal agent against Salmonella in milk and dairy products. Here, phage ZCSE6 has been isolated from raw milk sample sand molecularly and chemically characterized. At different multiplicities of infection (MOIs) of 0.1, 0.01, and 0.001, the phage–Salmonella interaction was studied for 6 h at 37 °C and 24 h at 8 °C. In addition, ZCSE6 was tested against Salmonella contamination in milk to examine its lytic activity for 3 h at 37 °C. The results showed that ZCSE6 has a small genome size (<48.5 kbp) and belongs to the Siphovirus family. Phage ZCSE6 revealed a high thermal and pH stability at various conditions that mimic milk manufacturing and supply chain conditions. It also demonstrated a significant reduction in Salmonella concentration in media at various MOIs, with higher bacterial eradication at higher MOI. Moreover, it significantly reduced Salmonella growth (MOI 1) in milk, manifesting a 1000-fold decrease in bacteria concentration following 3 h incubation at 37 °C. The results highlighted the strong ability of ZCSE6 to kill Salmonella and control its growth in milk. Thus, ZCSE6 is recommended as a biocontrol agent in milk to limit bacterial growth and increase the milk shelf-life.

Download Full-text

1667. Change in characteristics of community-onset ciprofloxacin-resistant E. coli isolates causing community-acquired acute pyelonephritis in South Korea

Open Forum Infectious Diseases ◽

10.1093/ofid/ofaa439.1845 ◽

2020 ◽

Vol 7 (Supplement_1) ◽

pp. S820-S820

Author(s):

Bongyoung Kim ◽

Ki Tae Kwon ◽

Seong-yeol Ryu ◽

Seong-Heon Wie ◽

Hyun-uk Jo ◽

...

Keyword(s):

Antimicrobial Resistance ◽

South Korea ◽

Molecular Characterization ◽

Acute Pyelonephritis ◽

Multilocus Sequence Typing ◽

Quinolone Resistance ◽

E Coli ◽

Extended Spectrum ◽

Community Onset

Abstract Background The aim of this study was to examine the change in characteristics of community-onset ciprofloxacin-resistant (CIP-R) E. coli isolates causing community-acquired acute pyelonephritis (CA-APN) in South Korea between 2010-2011 and 2017-2018. Methods E. coli samples isolated from the blood or urine were collected from patients with CA-APN aged 19 years and more who were admitted to 8 Korean hospitals from September 2017 to August 2018, prospectively. One isolate was collected from each patient. Phylogenetic typing, multilocus sequence typing (MLST), and molecular characterization of β-lactamase resistance and plasmid-mediated quinolone resistance (PMQR) determinants were performed. The data were compared with those from the previous study with same design in 2010-2011. Results A total of 346 and 300 isolates were collected during 2017-2018 and 2010-2011, respectively. Among them, 76 (22.0%) and 77 (25.7%) were CIP-R isolates. Significantly higher antimicrobial resistance against ampicillin (75.7% vs. 100%, P < 0.001) and cefotaxime (23.9% vs. 77.9%, P < 0.001) were observed for isolates in 2017-2018 compared to those in 2010-2011. The proportion of phylogenic group B2 had increased significantly (44.7% vs. 79.2%, P < 0.001). As for MLST, the proportion of ST131 (27.6% vs. 66.2%, P < 0.001) had increased while that of ST393 (18.4% vs. 3.9%, P =0.004) had decreased significantly. Higher proportion of CIP-R E. coli isolates in 2017-2018 had extended-spectrum β-lactamase (ESBL)/plasmid-mediated AmpC β-lactamase (PABL) (23.7% vs. 79.2%, P < 0.001) and PMQR determinant (11.8% vs. 40.8%, P < 0.001) compared to those in 2010-2011. Phlogenetic tree Analyzed by SplitsTree Conclusion Among uropathogenic CIP-R E. coli isolates in South Korea, ST131 predominance had become more prominent and the proportion of containing ESBL/PABL and/or PMQR determinants had increased. Disclosures All Authors: No reported disclosures

Download Full-text

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

Bangladesh Journal of Veterinary Medicine ◽

10.3329/bjvm.v14i2.31409 ◽

2017 ◽

Vol 14 (2) ◽

pp. 277-282 ◽

Cited By ~ 1

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 cows 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%).

Download Full-text

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

International Journal of Basic and Applied Sciences ◽

10.14419/ijbas.v5i2.5299 ◽

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

This study was conducted to detect Mycobacterium tuberculosis 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 Mycobacterium tuberculosis 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 Mycobacterium tuberculosis 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.

Download Full-text

Characterization of Extended-Spectrum β-Lactamase–Producing Escherichia coli Strains Isolated from Retail Foods in Shaanxi Province, China

Journal of Food Protection ◽

10.4315/0362-028x.jfp-14-490 ◽

2015 ◽

Vol 78 (5) ◽

pp. 1018-1023 ◽

Cited By ~ 14

Author(s):

MEILI XI ◽

QIAN WU ◽

XIN WANG ◽

BAOWEI YANG ◽

XIAODONG XIA ◽

...

Keyword(s):

Escherichia Coli ◽

Shaanxi Province ◽

People’S Republic Of China ◽

People's Republic Of China ◽

M Gene ◽

Disk Diffusion Test ◽

Republic Of China ◽

E Coli ◽

Extended Spectrum

Extended-spectrum β-lactamase (ESBL)–producing Escherichia coli strains have been reported worldwide; however, the incidence and characterization of foodborne ESBL-producing E. coli strains have been rarely reported in the People's Republic of China. Among a collection of 659 E. coli isolates recovered from retail foods in Shaanxi Province, People's Republic of China, 223 cefoxitin-resistant and/or cefoperazone-resistant isolates were screened for ESBL production with the double disk diffusion test. The ESBL-producing isolates were characterized for antimicrobial resistance and the presence of blaTEM, blaSHV, and blaCTX-M genes. Isolates with blaCTX-M were further classified by PCR as having blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, or blaCTX-M-25. One hundred forty-seven isolates were identified as ESBL positive. PCR detection revealed that 146 isolates (99.3%) contained the blaCTX-M gene. Among these isolates, 42 (28.8%) were positive for the enzyme CTX-M-1, 5 (3.4%) for CTX-M-2, and 99 (67.8%) for CTX-M-9. No CTX-M-8 and CTX-M-25 were found in this study. One hundred fifteen isolates (78.2%) were positive for the blaTEM gene, but blaSHV was not detected. Among the 147 ESBL-producing E. coli isolates, 75 (51.0%), 35 (23.8%), and 4 (2.7%) isolates were positive for blaTEM and blaCTX-M-9, blaTEM and blaCTX-M-1, and blaTEM and blaCTX-M-2, respectively. All of the 147 ESBL-producing isolates were resistant to three or more non–β-lactam antibiotics. This study provides evidence that foodborne E. coli can harbor ESBL-encoding genes. Thus, food could be a vehicle for the dissemination of ESBL-producing E. coli strains, a situation that requires surveillance and appropriate management strategies.

Download Full-text