Sensitive quantification of Clostridium perfringens

Clostridium perfringens is one of the most widely dispersed opportunistic pathogens and is well known to produce a number of toxins to cause several forms of histotoxic and enteric diseases in humans and animals [2]. Based on the production of four major toxins i.e., alpha, beta, epsilon and iota, it is categorized into five toxin-types viz. A, B, C, D and E. While it is ambiguous why C. perfringens produces so many diverse toxins, it is well known that it uses chromosomally-encoded α-toxin (which has phospholipase C (plc) and sphingomyelinase activities with hemolytic, necrotic and lethal abilities) as a chief virulent factor and key mediator for most of C. perfringens-associated diseases.”

Standard curves representing the quantitative detection of reference strains of C. perfringens by Amp-qPCR assay. C. perfringens ATCC 13124T, ATCC 9856, ATCC 3624, ATCC 3626, ATCC 12917, ATCC 14809, ATCC 27324, and CS 052–1 were cultivated separately in Glu-mGAM. DNA fractions were extracted from the culture samples in the early stationary phase (24 h), and bacterial counts were determined microscopically with DAPI staining. 10-fold serial dilutions of DNA corresponding to the bacterial counts ranging from 100 to 105 bacterial cells were assessed by 16S rRNA gene-specific aplc-specific b, and cpe-specific c Amp-qPCR assays. The Cq values obtained were plotted against the log10number of bacterial cells subjected to each reaction. Data are expressed as means and standard deviations of the results from 7 strains (ATCC 13124T, ATCC 9856, ATCC 3624, ATCC 3626, ATCC 12917, ATCC 14809, and ATCC 27324) in the 16S rRNA gene-specific and plc-specific primer sets, and 3 strains (ATCC 12917, ATCC 14809, and CS 052–1) in the cpe-specific primer set

Continue reading

 Bacterial translocation as a cause of surgical sepsis

‘The term ‘bacterial translocation’ is used to describe the passage of viable resident bacteria from the gastrointestinal tract to normally sterile tissues such as the mesenteric lymph nodes and other internal organs.7 The term also applies to the passage of inert particles and other macromolecules, such as lipopolysaccharide endotoxin, across the intestinal mucosal barrier.’

‘Bacterial translocation has been shown to occur in various patient populations.6 As already stated, it occurs in patients undergoing elective abdominal surgery, organ donors and those with intestinal obstruction, colorectal cancer, ischaemia–reperfusion injury shock and pancreatitis. Many authors suggest an increased prevalence in patients with obstructive jaundice, those receiving parenteral nutrition and the malnourished, but the evidence for this is limited. Interestingly, translocation, assessed by endotoxin or bacterial culture of portal or systemic blood, has only rarely been demonstrated after trauma’

Continue reading

Role of bacterial translocation in post op infection after GI surgery.

‘The incidence of postoperative sepsis has increased in the past decades, with the proportion of severe sepsis cases rising to unprecedented levels. Cases of sepsis are noted both after elective and emergency surgeries, but in the cases of elective surgeries, mortality is not respectively affected. Gastrointestinal perforation is the most common surgical condition requiring immediate surgical intervention. More specifically, colonic perforation may cause peritonitis through the spread of bacteria from the intestines, and, therefore, there is a high risk for further bacterial spread via blood flow.”

Figure 1 Mechanism of bacterial translocation after major gastrointestinal surgery.
Continue reading

Bacterial Translocation in Abdominal Trauma and Postoperative Infections

There is higher risk of BT in trauma patients, and it is associated with a significant increase of postoperative infections. An abdominal trauma index ≥10 was found to be associated with the development of BT. This is the first study describing BT among patients with abdominal trauma, where causality is confirmed at molecular level.

Continue reading

Bacterial translocation

‘Bacterial translocation (BT) describes the passage of bacteria from the gastrointestinal tract to normally sterile tissues such as the mesenteric lymph nodes (MLNs) and other internal organs. The clinical and pathophysiological significance of BT remains controversial. This report describes results obtained over a 13-year period of study.”

Table 1. Patients characteristics and indication for laparotomy

Type of surgery
 Emergency185
 Elective742
Diagnosis
 Malignancy543 (58·6)
  Upper gastrointestinal26
  Pancreaticohepatobiliary16
  Colorectal402
  Other99
 Inflammatory bowel disease94 (10·1)
  Crohn’s disease44
  Ulcerative colitis50
 Benign upper gastrointestinal54 (5·8)
 Benign pancreaticohepatobiliary44 (4·7)
 Benign lower gastrointestinal105 (11·3)
 Abdominal aortic aneurysm87 (9·4)
Continue reading