Patients suffering from acute calculous cholecystitis undergoing subacute laparoscopic cholecystectomy.

Samples of bile and gallbladder wall specimens will be taken during surgery and will be sent for microbiological and pathological investigation

Patients suffering from painattacks due to gallstones, who have not symptoms of cholecystitis. The patients will undergo elective laparoscopic cholecystectomy.

Samples of bile and gallbladder wall specimens will be taken during surgery and will be sent for microbiological and pathological investigation

Bile and gallbladder wall specimens will be taken from all patients included in the study. The material will be sent for microbiological and histopathological investigation.

Microbiological investigation of microorganisms present in the bile and in the gallbladder wall.

Bile samples will be collected by puncturing the gallbladder either perioperatively or immediate after extraction of the gallbladder from the abdomen. This is done in a sterile manner using a syringe.

The gallbladder wall specimens are collected immediately after extraction of the gallbladder from the abdomen. Three samples will be taken from macroscopic normal looking parts of the posterior wall in the fundus area using a sterile 4 mm punch biopsy.

Bile samples and punch biopsies sent to the Department of Clinical Microbiology, SLB, will be cultured on selected agar growth media and incubated under aerobic, microaerophilic, and anaerobic conditions.

Bacterial colonies will be identified and quantified using routine laboratory methods.

Microbiological investigation of antibiotic resistans of the bacteria, if any, present in bile and gallbladder wall specimens.

Antimicrobial susceptibility testing will be performed using the disk diffusion method.

To determine the possible presence of inhibitory concentrations of antibiotics in the bile samples, a droplet of bile from each sample will be added to an agar plate inoculated with the E. coli ATCC 25922-strain. After incubation, the appearance of a growth inhibition zone will be measured.

Histopathological investigation of the gallbladder to investigate if there is either inflammation or an actual infection and correlate these findings to the bacterial colonization.

The gallbladder from all included patients will be sent to the Department of Pathology, SLB, for evaluation of the grade of inflammation.

After formalin fixation sections from the cystic duct, the corpus and the fundus area will be sampled. The sections will be embedded in paraffin and will be cut into thin slices, which will be dyed with hematoxylin and eosin. The grade of acute inflammation will be assessed in a semi quantitative manner:

Grade 1: superficial ulceration of the mucosa concomitant with light infiltration by neutrophile granulocytes Grade 2: infiltration by granulocytes, spreading into the deeper layers of the gallbladder wall.

Grade 3: profound necrosis, extending through the entire gallbladder wall.