ABSTRACT
Clostridium perfringens is an anaerobic bacterium ubiquitous in various environments, especially in soil and the gastrointestinal tract of healthy humans and animals. In this study, multilocus sequence typing protocol was used to investigate genotypic relationships among 40 C. perfringens strains isolated from humans and broiler chicken with necrotic enteritis [NE]. The results indicated a few clonal populations, mainly observed in human strains, with 32.5% of all strains associated with one of three clonal complexes and 30 sequences types. The CC-1 cluster showed an interesting and unexpected result because it contained seven strains [six from animals and one of human origin]. Detection assays for toxin genes tpeL and netB were also performed. The netB gene was only observed in 7.5% of the strains from healthy human. The toxin gene tpeL was detected in 22.5% of the C. perfringens strains isolated from three individuals and in six broilers with NE. Our study describes the role of some C. perfringens strains of human origin acting as reservoirs of virulence genes and sources of infection. In addition, the strains of human and animal origin were found to be genetically distinct but phylogenetically close, and the human strains showed more diversity than the animal strains.
Subject(s)
Bacterial Proteins/genetics , Bacterial Toxins/genetics , Clostridium perfringens/classification , Clostridium perfringens/genetics , Enteritis/veterinary , Enterotoxins/genetics , Genotype , Multilocus Sequence Typing , Poultry Diseases/microbiology , Animals , Chickens , Child , Child, Preschool , Clostridium perfringens/isolation & purification , Cluster Analysis , Enteritis/microbiology , Genetic Variation , Healthy Volunteers , HumansABSTRACT
Childhood obesity is an increasing problem at the global level and considered as a risk factor for obesity development and the associated co-morbidities in adult life. In this study, the occurrence of Bacteroides fragilis group, Clostridium spp., Bifidobacterium spp. and Escherichia coli in 84 faecal samples from 30 obese, 24 overweight and 30 lean children was verified by culture technique and quantitative determination by quantitative PCR. In addition, Lactobacillus spp. and Methanobrevibacter smithii were also analysed. A correlation between the body mass index (BMI) and these bacteria was sought. Bacteroides vulgatus, Clostridium perfringens and Bifidobacterium adolescentis were most prevalent in all samples evaluated by culture-method. The B. fragilis group were found at high concentrations in obese and overweight children when compared with the lean ones (p 0.015). The obese and overweight children harboured higher numbers of Lactobacillus spp. than lean children (p 0.022). The faecal concentrations of the B. fragilis group (r = 0.24; p 0.026) and Lactobacillus spp. (r = 0.44; p 0.002) were positively correlated with BMI. Bifidobacterium spp. were found in higher numbers in the lean group than the overweight and obese ones (p 0.042). Furthermore, a negative correlation between BMI and Bifidobacterium spp. copy number (r = -0.22; p 0.039) was observed. Our findings show some difference in the intestinal microbial ecosystem of obese children compared with the lean ones and a significant association between number of Lactobacillus spp. and B. fragilis group and BMI.
Subject(s)
Body Mass Index , Feces/microbiology , Gastrointestinal Microbiome , Brazil/epidemiology , Child , Child, Preschool , Female , Humans , Male , Metagenome , Metagenomics/methods , Microbiota , Obesity/epidemiology , Obesity/etiology , RNA, Ribosomal, 16S/genetics , Risk FactorsABSTRACT
A rapid real-time PCR (RT-PCR) approach was developed to detect the bft gene subtypes in Bacteroides fragilis isolated from fecal samples. DNA obtained from diarrhea (110) and nondiarrhea (150) samples was evaluated. Subtype 1 was observed in 9 (8.2%) diarrhea and 7 (4.7%) nondiarrhea samples. Subtype 2 was not detected in any DNA samples, and subtype 3 was observed in only 1 diarrhea sample. The presence of the bft-1 gene did not show any statistically significant differences between the groups of children. This technique could be used to evaluate a possible correlation between disease and the presence of B. fragilis enterotoxin.
Subject(s)
Bacteroides Infections/microbiology , Bacteroides fragilis/classification , Bacteroides fragilis/isolation & purification , Diarrhea/microbiology , Bacterial Toxins/genetics , Bacteroides fragilis/genetics , Child , Child, Preschool , Female , Humans , Infant , Male , Metalloendopeptidases/genetics , Molecular Typing/methods , Polymerase Chain Reaction/methodsABSTRACT
Members of the genera Bacteroides and Parabacteroides are important constituents of both human and animal intestinal microbiota, and are significant facultative pathogens. In this study, the ability of Bacteroides spp. and Parabacteroides distasonis isolated from both diarrhoeal and normal stools (n = 114) to adhere to and invade HEp-2 cells was evaluated. The presence of putative virulence factors such as capsule and fimbriae was also investigated. Adherence to HEp-2 cells was observed in 75.4% of the strains, which displayed non-localized clusters. Invasion was observed in 37.5% and 26% of the strains isolated from diarrhoeal and non-diarrhoeal stools, respectively. All strains displayed a capsule, whereas none of them showed fimbriae-like structures. This is the first report of the ability of Bacteroides spp. and P. distasonis to adhere to and invade cultured HEp-2 epithelial cells.
Subject(s)
Bacterial Adhesion , Bacteroidetes/physiology , Bacteroidetes/pathogenicity , Diarrhea/microbiology , Gastrointestinal Tract/microbiology , Animals , Bacterial Capsules/analysis , Bacteroidetes/cytology , Bacteroidetes/isolation & purification , Cell Line , Child , Child, Preschool , Colony Count, Microbial , Cytosol/microbiology , Epithelial Cells/microbiology , Feces/microbiology , Fimbriae, Bacterial , Humans , Infant , Microscopy, Electron , Microscopy, Immunoelectron , Virulence Factors/analysisABSTRACT
The goals of this study were to evaluate if hemoglobin played a role as adjuvant in experimental peritonitis in horses and could cause clinical and haematological alterations that could be used for diagnosis and prognosis of cases of peritonitis. Fifteen adult horses were randomly divided into 5 equal groups, which were injected intraperitoneally with the following suspension: GI: 1x109 colony-forming units (CFU) of E. coli diluted in 500 mL of 0.9% saline solution plus 5 g of hemoglobin; GII: 1x109 CFU of B. fragilis diluted in 500 mL of 0.9% saline plus 5 g of hemoglobin; GIII: 1x109 CFU of E. coli in combination with 1x109 CFU of B. fragilis diluted in 500 ml of 0.9% of saline plus 5 g of hemoglobin; GIV: 500 mL of 0.9% saline plus 5 g of hemoglobin and GV: 500 mL of 0.9% saline. Leukopenia with neutropenia was observed in GI and GIII and a significant increase in plasma fibrinogen concentration occurred in horses of GI. There was a significant increase in total nucleated cell count in peritoneal fluid in horses of GI, GII, GIII and GIV. Fever, tachycardia, tachypnea, abdominal wall sensibility and tension, diarrhoea, colic, and decreased borborygmi sounds were the most frequent clinical signs observed in horses of GI, GII, GIII and GIV. In conclusion, hemoglobin was able to cause chemical peritonitis in horses, it had an adjuvant effect when associated t
ABSTRACT
The goals of this study were to evaluate if hemoglobin played a role as adjuvant in experimental peritonitis in horses and could cause clinical and haematological alterations that could be used for diagnosis and prognosis of cases of peritonitis. Fifteen adult horses were randomly divided into 5 equal groups, which were injected intraperitoneally with the following suspension: GI: 1x109 colony-forming units (CFU) of E. coli diluted in 500 mL of 0.9% saline solution plus 5 g of hemoglobin; GII: 1x109 CFU of B. fragilis diluted in 500 mL of 0.9% saline plus 5 g of hemoglobin; GIII: 1x109 CFU of E. coli in combination with 1x109 CFU of B. fragilis diluted in 500 ml of 0.9% of saline plus 5 g of hemoglobin; GIV: 500 mL of 0.9% saline plus 5 g of hemoglobin and GV: 500 mL of 0.9% saline. Leukopenia with neutropenia was observed in GI and GIII and a significant increase in plasma fibrinogen concentration occurred in horses of GI. There was a significant increase in total nucleated cell count in peritoneal fluid in horses of GI, GII, GIII and GIV. Fever, tachycardia, tachypnea, abdominal wall sensibility and tension, diarrhoea, colic, and decreased borborygmi sounds were the most frequent clinical signs observed in horses of GI, GII, GIII and GIV. In conclusion, hemoglobin was able to cause chemical peritonitis in horses, it had an adjuvant effect when associated t
ABSTRACT
The goals of this study were to evaluate if hemoglobin played a role as adjuvant in experimental peritonitis in horses and could cause clinical and haematological alterations that could be used for diagnosis and prognosis of cases of peritonitis. Fifteen adult horses were randomly divided into 5 equal groups, which were injected intraperitoneally with the following suspension: GI: 1x109 colony-forming units (CFU) of E. coli diluted in 500 mL of 0.9% saline solution plus 5 g of hemoglobin; GII: 1x109 CFU of B. fragilis diluted in 500 mL of 0.9% saline plus 5 g of hemoglobin; GIII: 1x109 CFU of E. coli in combination with 1x109 CFU of B. fragilis diluted in 500 ml of 0.9% of saline plus 5 g of hemoglobin; GIV: 500 mL of 0.9% saline plus 5 g of hemoglobin and GV: 500 mL of 0.9% saline. Leukopenia with neutropenia was observed in GI and GIII and a significant increase in plasma fibrinogen concentration occurred in horses of GI. There was a significant increase in total nucleated cell count in peritoneal fluid in horses of GI, GII, GIII and GIV. Fever, tachycardia, tachypnea, abdominal wall sensibility and tension, diarrhoea, colic, and decreased borborygmi sounds were the most frequent clinical signs observed in horses of GI, GII, GIII and GIV. In conclusion, hemoglobin was able to cause chemical peritonitis in horses, it had an adjuvant effect when associated t