scholarly journals Enterotoxin and Emetic Toxin Genes Profiles and Genetic Diversity of Bacillus cereus Isolated from Food, Environmental and Clinical Samples in Serbia

2020 ◽  
Vol 70 (2) ◽  
pp. 182-193
Author(s):  
Savić Dejana ◽  
Ristanović Elizabeta ◽  
Miljković Selimović Biljana ◽  
Radaković Sonja ◽  
Jošić Dragana ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Random Amplified Polymorphic Dna ◽  
Food Samples ◽  
Selective Medium ◽  
Clinical Samples ◽  
Environmental Isolates ◽  
Toxin Genes ◽  
Polymerase Chain ◽  
Systemic Infections ◽  
Diarrhea Syndrome

AbstractBacillus cereus, usually ingested by food, can cause two types of disease due to the presence of toxins: vomiting and diarrhea syndrome. Systemic infections can also occur. The aim was to detect genes for enterotoxins (hblA, entFM) and emetic toxin (cer) and to investigate the genetic heterogeneity of B. cereus isolates from food, environment and human stool. Identification of B. cereus was performed by means of selective medium, classical biochemical test and polymerase chain reaction (PCR). Toxin genes were detected by PCR. Typing was performed by random amplified polymorphic DNA (RAPD). EntFM gene was present in all stool and food samples and in 28/30 environmental isolates. HblA gene was present in 29/30 stool, 23/30 food and 24/30 environmental isolates. Cer gene was present in 30/30 stool, 28/30 food and 25/30 environmental isolates. The RAPD results show high heterogeneity among the isolates from each group. In the cumulative dendrogram, representative isolates from all three groups formed two clusters with a difference of 53%. The detection of toxin genes in all B. cereus isolates indicated these bacteria as potentially pathogenic and a serious threat for human health. The presence of isolates from all three groups in the same cluster suggests the existence of similar strains in the environment, food and patients, which is in line with the circulation of strains in nature through the food chain.

Toxin Genes Profiles and Toxin Production Ability of Bacillus cereus Isolated from Clinical and Food Samples

2011 ◽  
Vol 76 (1) ◽  
pp. T25-T29 ◽  
Author(s):  
Jung-Beom Kim ◽  
Jai-Moung Kim ◽  
Seung-Hak Cho ◽  
Hyuk-Soo Oh ◽  
Na Jung Choi ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Toxin Production ◽  
Food Samples ◽  
Toxin Genes

scholarly journals Antimicrobial susceptibility and β-lactamase production in Bacillus cereus isolates from stool of patients, food and environment samples

2016 ◽  
Vol 73 (10) ◽  
pp. 904-909 ◽  
Author(s):  
Dejana Savic ◽  
Biljana Miljkovic-Selimovic ◽  
Zorica Lepsanovic ◽  
Zoran Tambur ◽  
Sonja Konstantinovic ◽  
...  
Keyword(s):  
Bacillus Cereus ◽  
Antibiotic Susceptibility ◽  
Selective Medium ◽  
Biochemical Test ◽  
High Rate ◽  
Significant Difference ◽  
Disk Diffusion Assay ◽  
Systemic Manifestations ◽  
Polymerase Chain ◽  
Human Stool

Background/Aim. Bacillus cereus (B. cereus) usually ingested by food can cause two types of diseases: vomiting due to the presence of emetic toxin and diarrheal syndrome, due to the presence of diarrheal toxins. Systemic manifestations can also occur. The severe forms of disease demand antibiotic treatmant. The aim of this study was to determine the differences in antibiotic susceptibility and ?-lactamase activity of B. cereus isolates from stools of humans, food and environment. Methods. Identification of B. cereus was performed with selective medium, classical biochemical test and polymerase chain reaction (PCR) with primers specific for bal gene. Thirty isolates from each group were analysed for antibiotic susceptibility using the disk-diffusion assay. Production of ?-lactamase was determined by cefinase test, and double-disc method. Results. All strains identified as B. cereus using classical biochemical test, yielded 533 bp fragment with PCR. Isolates from all the three groups were susceptible to imipenem, vancomycin, and erythromycin. All isolates were susceptible to ciprofloxacin but one from the environment. A statistically significant difference between the groups was confirmed to tetracycline and trimethoprim-sulphamethoxazole sensitivity. A total of 28/30 (93.33%) samples from the foods and 25/30 (83.33%) samples from environment were approved sensitive to tetracycline, while 10/30 (33.33%) isolates from stools were sensitive. Opposite to this result, high susceptibility to trimethoprim-sulphamethoxazole was shown in samples from stools (100%), while isolates from foods (63.33%) and from environment (70%) had low susceptibility. All samples produced ?-lactamases. Conclusion. The strains of B. cereus from all the three groups showed high rate of sensitivity to most tested antibiotics, except to tetracycline in samples from human stool and to trimethoprim-sulphamethoxazole in samples from food and environment. The production of ?-lactamases was confirmed in all the strains.

Development and Use of Polymerase Chain Reaction for the Specific Detection of Salmonella Typhimurium in Stool and Food Samples

1999 ◽  
Vol 62 (10) ◽  
pp. 1103-1110 ◽  
Author(s):  
JER-SHENG LIN ◽  
HAU-YANG TSEN
Keyword(s):  
Polymerase Chain Reaction ◽  
Salmonella Typhimurium ◽  
Pcr Primers ◽  
Food Samples ◽  
Clinical Samples ◽  
Specific Detection ◽  
Chain Reaction ◽  
Pcr Product ◽  
Polymerase Chain ◽  
Salmonella Serovars

Salmonella Typhimurium is one of the most important Salmonella serovars that may cause foodborne disease and human salmonellosis infection. Detection of this organism in the clinical samples of persons with gastroenteritis and the food samples associated with such persons may allow us to trace the cause of disease. Because malic acid dehydrogenase, an enzyme of the citric acid cycle, is common to organisms, the gene (mdh) coding for this enzyme was selected for the design of Salmonella Typhimurium-specific polymerase chain reaction (PCR) primers. By comparison of the mdh gene sequences of Salmonella Typhimurium and other Salmonella serotypes and of some isolates of other genera, two oligonucleotides were designed and used as PCR primers for the specific detection of Salmonella Typhimurium. The molecular weight of the PCR product was 261 bp as expected. Salmonella serovars other than Salmonella Typhimurium and isolates of other genera in the Enterobacteriaceae that is closely related to Salmonella did not generate any false-positive results. When this primer pair was used for the detection of Salmonella Typhimurium cells artificially inoculated into human stool specimens and food samples, such as milk and raw chicken meat, levels as low as 100 CFU per 0.1 g of stool specimen or per ml of milk or food homogenate could be detected if an 8- to 12-h preculture step using combined lactose-tetrathionate broth was performed prior to the PCR.

Distribution of Iron Uptake Systems Encoding Genes Among the Clinical Isolates of Escherichia coli Compared to Foodstuffs Isolates

2020 ◽  
Vol 20 (4) ◽  
pp. 517-522
Author(s):  
Hassan Mahmoudi ◽  
Hadi Hossainpour ◽  
Mohammad Moradi ◽  
Mohammad Yousef Alikhani
Keyword(s):  
Escherichia Coli ◽  
Iron Uptake ◽  
Food Poisoning ◽  
Food Samples ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
E Coli ◽  
Microbiological Methods ◽  
Polymerase Chain ◽  
Encoding Genes

Background: Bacteria require iron ions to grow and infect the host, which, by using iron uptake systems, acquire free iron from their host cell. Escherichia coli is one of the most important pathogens to cause food poisoning and clinical infections. The aim of this study was to assess the distribution of iron uptake systems encoding genes in clinical isolates of E.coli compared to food samples isolates. Materials and Methods: This investigation was conducted to determine the prevalence of E. coli isolated from various sources of food and clinical specimens. The E. coli isolates confirmed by the standard microbiological methods. The isolates were examined for the presence of iut A and iuc A genes by specific primers using the polymerase chain reaction technique. Results: A total of 100 and 50 isolates of E. coli were collected from clinical samples and foodstuffs, respectively. The prevalence of E. coli in the food and clinical samples was 33.33% and 64.10%, respectively. The frequency of iut A and iuc A genes in the food and clinical isolates were 76%-84% and 86% - 83%, respectively. Conclusion: Our results showed that the prevalence of E. coli isolates with iut A and iuc A genes was relatively higher compared to many previous studies. The existence of these genes in E. coli strains is likely to be related to pathogenicity in those strains, which requires further studies in the future.

An improved selective and diagnostic medium for the isolation and enumeration of Bacillus cereus in foods

1980 ◽  
Vol 26 (7) ◽  
pp. 753-759 ◽  
Author(s):  
R. Holbrook ◽  
Judith M. Anderson
Keyword(s):  
Bacillus Cereus ◽  
Egg Yolk ◽  
Bromothymol Blue ◽  
Food Samples ◽  
Selective Medium ◽  
Confirmatory Test ◽  
Staining Procedure ◽  
Complete Agreement ◽  
Phenol Red ◽  
Biochemical Tests

The use and performance of an improved diagnostic and selective medium, PEMBA (polymyxin pyruvate egg yolk mannitol bromothymol blue agar), for the detection of Bacillus cereus in foods is described. The distinct colonial appearance of B. cereus on PEMBA permitted the recognition of both strains: those that do precipitate egg yolk and those that do not react with egg yolk. A staining procedure, used to demonstrate microscopically both the presence of lipid globules in vegetative cells and spore morphology of isolates, proved a rapid and reliable confirmatory test which gave complete agreement with a battery of biochemical tests used for this purpose. The quantitative recovery of B. cereus on PEMBA from 143 food samples was not significantly different from counts on KG (Kim and Goepfert), MYP (mannitol egg yolk phenol red), and McClung's media, and the selectivity of PEMBA was generally superior.

scholarly journals Identification of Fusarium oxysporum f. sp. basilici Isolated from Soil, Basil Seed, and Plants by RAPD Analysis

Plant Disease ◽  
1999 ◽  
Vol 83 (6) ◽  
pp. 576-581 ◽  
Author(s):  
Annalisa Chiocchetti ◽  
Stefano Ghignone ◽  
Andrea Minuto ◽  
M. Lodovica Gullino ◽  
Angelo Garibaldi ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Dna Polymerase ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Selective Medium ◽  
Chain Reaction ◽  
Rapd Pcr ◽  
Polymerase Chain ◽  
Clear Differentiation ◽  
Pathogenicity Assay

Fifty-two isolates of Fusarium oxysporum, obtained from infected basil plants, seed, flower residues, and soil from different growing areas in Italy and Israel, were analyzed by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), coupled to a DNA extraction protocol from colonies grown on Fusarium-selective medium. In a pathogenicity assay, 35 isolates caused 32 to 92% disease on seedlings of the highly susceptible basil cultivar Fine verde, while 17 isolates were nonpathogenic on basil. Thirty of the F. oxysporum f. sp. basilici isolates obtained from soil or wilted plants gave identical amplification patterns using 31 different random primers. All tested primers allowed clear differentiation of F. oxysporum f. sp. basilici from representatives of other formae speciales and from nonpathogenic strains of F. oxysporum. RAPD profiles obtained from DNA of isolates extracted directly from cultures grown on Fusarium selective medium were identical to those obtained from DNA extracted from lyophilized mycelia.

Investigation of duck plague virus in hoar areas of Bangladesh

2020 ◽  
Vol 26 (1-2) ◽  
pp. 73-78
Author(s):  
A Hossen ◽  
MH Rahman ◽  
MZ Ali ◽  
MA Yousuf ◽  
MZ Hassan ◽  
...  
Keyword(s):  
Cytopathic Effect ◽  
Clinical Sample ◽  
Chorioallantoic Membrane ◽  
Clinical Samples ◽  
Isolation And Identification ◽  
Duck Plague Virus ◽  
Pcr Method ◽  
Polymerase Chain ◽  
The Individual ◽  
Free Ranging

Duck plague (DP) is the most important infectious disease of geese, ducks and free-ranging water birds. The present study was conducted to determine the prevalence of duck plague virus followed by isolation and identification. For these purposes, a total of 155 cloacal swabs samples were collected randomly from duck of different haor areas of Bangladesh including 45 (41 surveillance and 4 clinical) samples from Netrokona; 42 (40 surveillance and 2 clinical) samples from Kishoregonj; 30 samples from Brahmanbaria and 38 samples from Sunamganj. The samples were processed and pooled (1:5 ratio) for initial screening of target polymerase gene of duck plague virus by polymerase chain reaction (PCR) method. All the samples of a positive pool were then tested individually for identifying the individual positive samples. The result showed that out of 155 samples, 41 (26.45%) were found positive in which 17 were from Netrokona, where 15 (36.58%) were from surveillance samples and 2 (50%) were from clinical sample; 16 were from Kishoregonj, where 14 (35%) were from surveillance samples and 2 (100%) were from clinical sample; 2 (6.6%) were from Brahmanbaria and 5 (13.15%) were from Sunamganj. These positive samples were inoculated into 9-10 days embryonated duck eggs (EDE) through chorioallantoic membrane (CAM) route for the isolation of virus. The EDE died earlier was also chilled, and in a similar way, the CAMs were collected and again performed PCR for id entification of virus. Out of 41 PCR positive samples, 26 samples were isolated and reconfirmed by PCR. Subsequently, DPV was isolated in primary duck embryo fibroblasts cell culture and confirmed by observing cytopathic effect (CPE). Bang. J. Livs. Res. Vol. 26 (1&2), 2019: P. 73-78

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