scholarly journals Isolation of enterotoxigenic Staphylococcus aureus harboring seb gene and enteropathogenic Escherichia coli (serogroups O18, O114, and O125) from soft and hard artisanal cheeses in Egypt

2020 ◽  
Vol 10 (3) ◽  
Author(s):  
Ola Wagih Hegab ◽  
Eman F. Abdel-Latif ◽  
Ashraf Ahmed Moawad

Background: Soft and hard artisanal cheeses are regularly consumed in Egypt. These products are usually processed from raw milk which may  harbor many pathogenic and spoilage microorganisms.Aim: To evaluate the safety of some artisanal cheeses in Egypt, such as Ras, Domiati, and Mish, through chemical and microbiological examination.Methods: One hundred and fifty random samples of traditional Ras, Domiati, and Mish cheeses (50 each) were microbiologically and chemically analyzed. Counts of total bacteria, presumptive coliform, staphylococci, yeast, and mold were estimated. Furthermore, isolation of Escherichia coli and Staphylococcus aureus was performed, followed by PCR confirmation; isolates of E. coli were examined for the presence of virulence genes; on the other hand, the detection of the five classical enterotoxin genes of S. aureus was performed using multiplex PCR. Regarding chemical analysis, moisture, salt, and acidity content were measured. Correlations between chemical and microbial findings were investigated.Results: Mean counts of total bacteria, presumptive coliform, staphylococci, yeast, and mold were (2 × 108, 3 × 106 and 1 × 107 ), (3 × 105, 5 × 10 and 5 × 102), (1 × 106, 4 × 105 and 1 × 105), (3 × 105, 1 × 105 and 5 × 105), and (7 × 103, 4 × 103 and 3 × 104) for Ras, Domiati and Mish cheeses,  respectively. Serological identification of suspected E. coli revealed that E. coli O125 was isolated from Ras and Domiati samples, E. coli O18 was recovered from Ras samples, while E. coli O114 was isolated from Mish samples. PCR results revealed that all detected isolates of E. coli were positive for both iss (increased serum survival) and fimH (type 1 fimbriae) genes. Concerning isolated S. aureus, all examined products were harboring S. aureus enterotoxigenic strains, with seb and sed genes being the most common. The mean values of moisture, salt, and acidity were (30.03, 56.44, and 58.70), (3.30, 6.63, and 7.56) and (0.65, 0.68, and 0.50) for Ras, Domiati, and Mish cheeses, respectively.Conclusion: Enterotoxigenic S. aureus harboring seb gene and enteropathogenic E. coli (serogroups O18, O114, and O125) were frequently isolated from soft and hard artisanal cheeses in Egypt. Therefore, strict hygienic measures should be applied during their manufacture, handing, and distribution. Keywords: Domiati, E. coli virulence genes, Mish, Ras, S. aureus enterotoxins.

2019 ◽  
Vol 40 (1) ◽  
pp. 163 ◽  
Author(s):  
Leandro Parussolo ◽  
Ricardo Antônio Pilegi Sfaciotte ◽  
Karine Andrezza Dalmina ◽  
Fernanda Danielle Melo ◽  
Ubirajara Maciel Costa ◽  
...  

The serrano artisanal cheese is a typical product from South region of Brazil, which is produced by skilled cheesemakers using raw milk. The contamination of this food by Escherichia coli has a great impact on public health, since it could threat the consumers’ health. The study evaluated the presence of virulence genes, antimicrobial susceptibility profiles and bofilm-production ability of Escherichia coli isolates obtained from raw milk and artisanal cheese produced in Southern Brazil. A total of 117 isolates of E. coli were characterized by multiplex PCR to detect the following virulence genes: eae for enteropatogenic E. coli (EPEC), lt and st for enterotoxigenic E. coli (ETEC), stx for shiga toxin-producing E. coli (STEC), stx and eae for enterohemorrhagic E. coli (EHEC), ipaH for enteroinvasive E. coli (EIEC) and aggR for enteroaggregative E. coli (EAEC). In addition, antimicrobial susceptibility profile to 22 antimicrobial agents was also performed by disk diffusion method, and we searched for extended-spectrum beta-lactamases (ESBL) and/or carbapenemase- producing isolates. Isolates that were positive for ESBL and carbapenemase were further investigated for the presence of the genes: blaTEM, blaSHV, blaOXA, blaCTX-M, for ESBL and blaOXA-48 for carbapenemase. Further, isolates had their ability to form biofilms investigated by the red Congo agar method. Virulence genes of E. coli were identified in 21.37% of the tested isolates, which were classified as EPEC (the most prevalent pathotype) and ETEC or EAEC. Ten (8.55%) of the total studied E. coli isolates revealed a multidrug-resistant profile, since they were resistant to three or more antimicrobial classes; whereas four isolates (3.42%) were classified as ESBL-producers and showed the presence of blaTEM gene. None of the isolates exhibited carbapenemase activity nor did they carry carbapenemase genes. From the total of E. coli isolates, 79 (67.52%) were considered potential biofilm producers. These results address a serious public health issue, since artisanal cheeses pose a risk to consumers’ health, since may be sources of dissemination of diarrheogenic E. coli, that can cause from subclinical to severe and fatal infections in children and adults, and also emphasize the need to improve adaptations/adjustments in the manufacturing processes of these products.


2020 ◽  
Vol 49 (1) ◽  
pp. 32-39 ◽  
Author(s):  
A. Vásquez García ◽  
S.H. Gomes de Sá ◽  
G. de Sousa Silva ◽  
J.E. Mejia Ballesteros ◽  
E. Barbieri ◽  
...  

The objective of this study was to evaluate the microbiological quality of oysters and mussels grown in Cananéia, Brazil, by analysing mesophiles, psychrothophic bacteria, moulds and yeasts, Staphylococcus aureus, and Salmonella spp., and to compare the efficiency of Compact Dry EC method and the conventional method for counting of total coliforms and Escherichia coli. The microbial analysis showed that the mean values of mesophilic counts were 3.14±0.81 log CFU g−1 for oysters and 3.92±0.90 for mussels; the mean values of psychrophilic counts were 2.78±0.75 log CFU g−1 for oysters and 3.22±0.75 log CFU g−1 for mussels; the mean values of mould and yeast counts were 3.70±0.58 log CFU g−1 for oysters and 3.33±0.81 log CFU g−1 for mussels. Salmonella spp. did not present positive results, and the maximal count of Staphylococcus aureus was 1.7 log CFU g−1, therefore, within the limits established in the legislation. The correlation coefficients between the Compact Dry EC method and conventional method were >0.87 for total coliform and E. coli counts for both types of shellfish. The data in this study show that the Compact Dry EC method is an acceptable alternative to conventional methods for enumeration of total coliforms and E. coli in shellfish.


2018 ◽  
Vol 48 (2) ◽  
Author(s):  
Fernanda Morcatti Coura ◽  
Amanda Nadia Diniz ◽  
Carlos Augusto Oliveira Junior ◽  
Andrey Pereira Lage ◽  
Francisco Carlos Faria Lobato ◽  
...  

ABSTRACT: This study identified the virulence genes, pathovars, and phylogenetic groups of Escherichia coli strains obtained from the feces of dogs with and without diarrhea. Virulence genes and phylogenetic group identification were studied using polymerase chain reaction. Thirty-seven E. coli isolates were positive for at least one virulence factor gene. Twenty-one (57.8%) of the positive isolates were isolated from diarrheal feces and sixteen (43.2%) were from the feces of non-diarrheic dogs. Enteropathogenic E. coli (EPEC) were the most frequently (62.2%) detected pathovar in dog feces and were mainly from phylogroup B1 and E. Necrotoxigenic E. coli were detected in 16.2% of the virulence-positive isolates and these contained the cytotoxic necrotizing factor 1 (cnf1) gene and were classified into phylogroups B2 and D. All E. coli strains were negative for the presence of enterotoxigenic E. coli (ETEC) enterotoxin genes, but four strains were positive for ETEC-related fimbriae 987P and F18. Two isolates were Shiga toxin-producing E. coli strains and contained the toxin genesStx2 or Stx2e, both from phylogroup B1. Our data showed that EPEC was the most frequent pathovar and B1 and E were the most common phylogroups detected in E. coli isolated from the feces of diarrheic and non-diarrheic dogs.


2018 ◽  
Vol 12 (07) ◽  
pp. 533-541 ◽  
Author(s):  
Walid Elmonir ◽  
Etab Abo-Remela ◽  
Azza Sobeih

Introduction: Milk is an important food in Egypt and most of it is sold as raw milk in informal markets. Methodology: This study was conducted to investigate the public health risks of Escherichia coli and Staphylococcus aureus in milk sold in informal markets in Egypt. A total of 121 milk samples were analyzed for occurrence, virulence genes and antibiotic resistance of E. coli and S. aureus. Results: A total of 35/121 (28.9%) of milk samples were contaminated with 16/121 (13.2%) E. coli, 22/121 (18.2%) S. aureus, and 3/121 (2.5%) both isolates. Shiga-toxin producing E. coli (STEC), Enterotoxigenic E. coli (ETEC) and Enterotoxigenic S. aureus were detected in 5/121 (4.1%), 2/121 (1.7%) and 8/121 (6.6%) of the examined milk samples, respectively. Multiple drug resistances (MDRs) were showed by 14/16 (87.5%) and 21/22 (95.5%) of E. coli and S. aureus isolates, respectively. E. coli isolates showed high resistance for cephalothin (87.5%), ampicillin (68.8%) and tetracycline (68.8%), but were sensitive for gentamicin and chloramphenicol. Resistance phenotypes of E. coli were diverse; however, STEC isolates were significantly associated with co-resistance to cephalothin, ampicillin and tetracycline (P< 0.05). Two (9.1%) of S. aureus isolates were methicillin-resistant (MRSA) but sensitive to gentamicin (GS-MRSA). Five (22.7%) of S. aureus isolates were gentamicin-resistant methicillin-sensitive S. aureus (GR-MSSA). S. aureus isolates also showed high resistance for ampicillin (100%), tetracycline (90.1%) and sulfamethoxazole-trimethoprim (90.1%). Conclusion: These findings highlighted the potential public health hazards of E. coli and S. aureus pathogens in raw milk sold in informal markets in Egypt.


2021 ◽  
Vol 12 ◽  
Author(s):  
Huimin Liu ◽  
Lu Meng ◽  
Lei Dong ◽  
Yangdong Zhang ◽  
Jiaqi Wang ◽  
...  

Escherichia coli is a common bacterium in the intestines of animals, and it is also the major important cause of toxic mastitis, which is an acute or peracute disease that causes a higher incidence of death and culling of cattle. The purpose of this study was to investigate E. coli strains isolated from the raw milk of dairy cattle in Northern China, and the antibacterial susceptibility of these strains and essential virulence genes. From May to September 2015, 195 raw milk samples were collected from 195 dairy farms located in Northern China. Among the samples, 67 (34.4%) samples were positive for E. coli. About 67 E. coli strains were isolated from these 67 samples. The prevalence of Shiga toxin-producing E. coli (STEC), enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), and enteroinvasive E. coli (EIEC) were 9, 6, 4.5, and 1.5%, respectively. Among the virulence genes detected, stx1 was the most prevalent (6/67, 9%) gene, followed by eae (3/67, 4.5%), and estB (2/67, 3%). Moreover, the strains exhibited different resistance levels to ampicillin (46.3%), amoxicillin-clavulanic acid (16.4%), trimethoprim-sulfamethoxazole (13.4%), tetracycline (13.4%), cefoxitin (11.9%), chloramphenicol (7.5%), kanamycin (7.5%), streptomycin (6.0%), tobramycin (4.5%), azithromycin (4.5%), and ciprofloxacin (1.5%). All of the E. coli isolates were susceptible to gentamicin. The prevalence of β-lactamase-encoding genes was 34.3% in 67 E. coli isolates and 45% in 40 β-lactam-resistance E. coli isolates. The overall prevalence of blaSHV, blaTEM, blaCMY, and blaCTX-M genes were 1.5, 20.9, 10.4, and 1.5%, respectively. Nine non-pathogenic E. coli isolates also carried β-lactamase resistance genes, which may transfer to other pathogenic E. coli and pose a threat to the farm’s mastitis management projects. Our results showed that most of E. coli were multidrug resistant and possessed multiple virulence genes, which may have a huge potential hazard with public health, and antibiotic resistance of E. coli was prevalent in dairy herds in Northern China, and ampicillin should be used cautiously for mastitis caused by E. coli in Northern China.


2020 ◽  
Vol 8 (3) ◽  
pp. 82-86
Author(s):  
Shyamala Rai ◽  
Barsha Karki ◽  
Sujita Humagain ◽  
Sandesh Rimal ◽  
Sandhya Adhikari ◽  
...  

The emergence of antibiotic resistance in microorganisms and the presence of such isolates in milk pose a great risk to public health. Therefore, this study aims to determine the antibiotic susceptibility pattern of Escherichia coli and Staphylococcus aureus isolated from milk and assess the microbial quality of milk. For this, a total of 70 milk samples were collected and the total bacterial count (TBC) was determined. E. coli and S. aureus were isolated using their respective selective media while antibiotic susceptibility testing was carried out by Kirby Bauer Disc Diffusion method. The TBC showed that the raw milk samples contained two-fold higher microbial load while the pasteurized milk samples contained four-fold higher microbial loads than the standard guidelines. A total of 62 isolates were identified from culture-positive milk samples of which 32 were E. coli and 30 were S. aureus. A significant correlation was observed between microbial load and the organism isolated (r = 0.339, p<0.01). All S. aureus isolates were susceptible to Chloramphenicol while 40% were resistant to Cefoxitin, indicating the presence of Methicillin resistant S. aureus (MRSA). Also, 12 multidrug resistant (MDR) S. aureus were identified. While for E. coli, all were susceptible to Chloramphenicol but resistant to Ampicillin. Also, 9 MDR E. coli were detected. Higher resistance was observed among isolates from the raw milk samples than the pasteurized milk. It can be concluded that the milk produced by small-scale farms and dairy industries of Kathmandu district are of poor quality. Hence, routine microbial quality assessment and antimicrobial resistance monitoring should be followed to safeguard public health.


Author(s):  
Mehtap Çiftçi ◽  
Nilgün Öncül

Since coliform bacteria are common both in the intestine and in nature (soil, plant, etc.), they are considered as a sanitation indicator in the food industry. It is known that the majority of bacteria defined as fecal coliform in the coliform group are Escherichia coli. The presence of E. coli or fecal coliform bacteria in any sample is an indication that the necessary hygienic measures are not taken during production, storage, and sale. That means the sample is directly or indirectly contaminated with faeces, and/or other intestinal pathogens may also exist. In this study, raw milk, freshly squeezed fruit juices, unpackaged ice cream, shaved ice, and ice-cold samples were purchased from the famous touristic destination Fethiye and analyzed for coliform bacteria. For this purpose, the samples were purchased from local marketplaces, buffets, cafes, patisseries, restaurants, and roadsides at Fethiye. In total 60 samples were analyzed using Violet Red Bile (VRB) Agar. The results of coliform bacteria ranged


2018 ◽  
Vol 41 (4) ◽  
pp. 353-363
Author(s):  
Alberto J. Valencia-Botin ◽  
Melesio Gutiérrez-Lomelí ◽  
Juan A. Morales-Del-Río ◽  
Pedro J. Guerrero-Medina ◽  
Miguel A. Robles-García ◽  
...  

Actualmente existe la necesidad de hacer frente al problema de la resistencia a los antibióticos y al uso indiscriminado de fungicidas químicos en la agricultura. El objetivo de este trabajo fue evaluar el efecto inhibitorio de extractos acuosos, metanólicos, acetónicos y hexánicos de hoja y tallo de Vitex mollis Kunth (Lamiaceae) contra diferentes bacterias (Escherichia coli, Micrococcus luteus, Salmonella enterica y Staphylococcus aureus) y especies del hongo Fusarium (F. verticillioides, F. oxysporum, F. tapsinum y F. oxysporum f.sp. lycopersici) de importancia en la salud y en la agricultura, así como determinar su composición química general. Se determinaron las concentraciones inhibitorias mínimas (CIM) de todos los extractos por la técnica de microdilución, excepto del hexánico, que no presentó inhibición en las bacterias estudiadas. S. enterica fue la bacteria que mostró mayor sensibilidad al extracto metanólico de tallo (CIM = 28 μg mL-1), le siguieron M. luteus (CIM = 32 μg mL-1), S. aureus (CIM = 75 μg mL-1) y E. coli (CIM = 80 μg mL- 1). Los extractos metanólicos y acuosos de tallo presentaron mayor porcentaje de inhibición contra los diferentes tipos de Fusarium evaluados por el método de dilución en agar. Los extractos de V. mollis inhibieron a F. verticillioides entre 62 y 91 % con 120 μg mL-1 de extracto. El orden de las especies de hongos inhibidas por los extractos fue: F. verticillioides > F. oxysporum > F. tapsinum > F. oxysporum f.sp. lycopersici. La composición química de las especies se determinó mediante pruebas para fenoles, taninos, flavonoides, triterpenos, alcaloides, cumarinas y saponinas. Ninguno de los extractos presentó alcaloides y saponinas. Los fenoles (37.1 mg EAG/g muestra seca) y flavonoides (26.8 mg EQ/g muestra seca) fueron los compuestos mayoritarios en los extractos metanólicos y acuosos. En conclusión, se requieren cantidades muy pequeñas de extracto para la inhibición de bacterias y de Fusarium; por lo tanto, V. mollis puede ser considerada una fuente de metabolitos para este fin y en la agricultura como control alternativo dentro de un manejo integrado de enfermedades.


2020 ◽  
Vol 24 (19) ◽  
pp. 2272-2282
Author(s):  
Vu Ngoc Toan ◽  
Nguyen Minh Tri ◽  
Nguyen Dinh Thanh

Several 6- and 7-alkoxy-2-oxo-2H-chromene-4-carbaldehydes were prepared from corresponding alkyl ethers of 6- and 7-hydroxy-4-methyl-2-oxo-2H-chromen-2-ones by oxidation using selenium dioxide. 6- and 7-Alkoxy-4-methyl-2H-chromenes were obtained with yields of 57-85%. Corresponding 4-carbaldehyde derivatives were prepared with yields of 41-67%. Thiosemicarbazones of these aldehydes with D-galactose moiety were synthesized by reaction of these aldehydes with N-(2,3,4,6-tetra-O-acetyl-β-Dgalactopyranosyl) thiosemicarbazide with yields of 62-74%. These thiosemicarbazones were screened for their antibacterial and antifungal activities in vitro against bacteria, such as Staphylococcus aureus, Escherichia coli, and fungi, such as Aspergillus niger, Candida albicans. Several compounds exhibited strong inhibitory activity with MIC values of 0.78- 1.56 μM, including 8a (against S. aureus, E. coli, and C. albicans), 8d (against E. coli and A. niger), 9a (against S. aureus), and 9c (against S. aureus and C. albicans).


2020 ◽  
Vol 15 (6) ◽  
pp. 665-679
Author(s):  
Alok K. Srivastava ◽  
Lokesh K. Pandey

Background: [1, 3, 4]oxadiazolenone core containing chalcones and nucleosides were synthesized by Claisen-Schmidt condensation of a variety of benzaldehyde derivatives, obtained from oxidation of substituted 5-(3/6 substituted-4-Methylphenyl)-1, 3, 4-oxadiazole-2(3H)-one and various substituted acetophenone. The resultant chalcones were coupled with penta-O-acetylglucopyranose followed by deacetylation to get [1, 3, 4] oxadiazolenone core containing chalcones and nucleosides. Various analytical techniques viz IR, NMR, LC-MS and elemental analysis were used to confirm the structure of the synthesised compounds.The compounds were targeted against Bacillus subtilis, Staphylococcus aureus and Escherichia coli for antibacterial activity and Aspergillus flavus, Aspergillus niger and Fusarium oxysporum for antifungal activity. Methods: A mixture of Acid hydrazides (3.0 mmol) and N, Nʹ- carbonyl diimidazole (3.3 mmol) in 15 mL of dioxane was refluxed to afford substituted [1, 3, 4]-oxadiazole-2(3H)-one. The resulted [1, 3, 4]- oxadiazole-2(3H)-one (1.42 mmol) was oxidized with Chromyl chloride (1.5 mL) in 20 mL of carbon tetra chloride and condensed with acetophenones (1.42 mmol) to get chalcones 4. The equimolar ratio of obtained chalcones 4 and β -D-1,2,3,4,6- penta-O-acetylglucopyranose in presence of iodine was refluxed to get nucleosides 5. The [1, 3, 4] oxadiazolenone core containing chalcones 4 and nucleosides 5 were tested to determined minimum inhibitory concentration (MIC) value with the experimental procedure of Benson using disc-diffusion method. All compounds were tested at concentration of 5 mg/mL, 2.5 mg/mL, 1.25 mg/mL, 0.62 mg/mL, 0.31 mg/mL and 0.15 mg/mL for antifungal activity against three strains of pathogenic fungi Aspergillus flavus (A. flavus), Aspergillus niger (A. niger) and Fusarium oxysporum (F. oxysporum) and for antibacterial activity against Gram-negative bacterium: Escherichia coli (E. coli), and two Gram-positive bacteria: Staphylococcus aureus (S. aureus) and Bacillus subtilis(B. subtilis). Result: The chalcones 4 and nucleosides 5 were screened for antibacterial activity against E. coli, S. aureus and B. subtilis whereas antifungal activity against A. flavus, A. niger and F. oxysporum. Compounds 4a-t showed good antibacterial activity whereas compounds 5a-t containing glucose moiety showed better activity against fungi. The glucose moiety of compounds 5 helps to enter into the cell wall of fungi and control the cell growth. Conclusion: Chalcones 4 and nucleosides 5 incorporating [1, 3, 4] oxadiazolenone core were synthesized and characterized by various spectral techniques and elemental analysis. These compounds were evaluated for their antifungal activity against three fungi; viz. A. flavus, A. niger and F. oxysporum. In addition to this, synthesized compounds were evaluated for their antibacterial activity against gram negative bacteria E. Coli and gram positive bacteria S. aureus, B. subtilis. Compounds 4a-t showed good antibacterial activity whereas 5a-t showed better activity against fungi.


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