ESCMID anaerob course, Groningen, September 29th to October 1st, 2014

- Anaerobes did and do not get a lot of attention
- Perhaps because there were not many resistance and the isolation was difficult
- But the resistance is increasing
- This course teach us on the importance of anaerobes in the clinic, the types of anaerobes
- The identification of anaerobes using biochemical techniques was part of the practical session
- The use of MALDI TOF in the identification was also discussed
- Possible use of new kids on the block such as Next Gen Sequencing was described
- Microbiomes and their roles in health were presented --> it is good to broaden our knowledge

Also....
- I like to go back to The Netherlands (I missed 'karnemelk' but not the 'broodjeslunch')....
- During the course, at September 30, there was an EARTHQUAKE in Groningen, 2.8 on the Richter scale. Oeps...

The potential use of microcalorimetry in rapid differentiation between septic arthritis and other causes of arthritis.

- Septic arthritis need to be recgnized early so timely treatment can be put into place
- We tested the use of microcalorimetry, a test that can measure the flow of heat sensitively on the synovial fluid
- We compared the results with conventional microbiological techniques such as Gram stain
- We showed that microcalorimetry was accurate and fast in differentiating septic arthritis from other conditions of arthritis
- The abstract of this paper can be found here 

Prolonged vs intermittent infusion of piperacillin/tazobactam in critically ill patients

- In critically ill patients, physiologic changes occur

- Piperacillin/ tazobactam is antibiotic where time above MIC feature is important for bacterial killing

- We described the rational of continous infusion of piperacillin/ tazobactam

- We also perfromed a systematic review comparing continous with other mode of infusion of this antibiotic specifically in critically care patients

- We showed that piperacillin/ tazobactam showed somehow benefit above other mode of infusion, but the level of evidence is only moderate

- The abstract of the paper can be found here.

ESCMID Antimicrobial Susceptibility Test course, Linz, September 16 to 19th, 2014

What is about
- We all ever heard about EUCAST and CLSI in determining whether bacteria are susceptible, intermediate or resistance to specific antibiotics
- This course explained how EUCAST come to the break points. Good to refresh the PK/ PD again and good to know what is an ECOFF!
- All types of antibiotic susceptibility tests (ASTs) were revisited. Not many of us have done microdilution for example.
- The role of AST in surveillance was explained

What I like about the course
- We do it all the day but no one really explained the rational behind it
- Fantastic organization
- Warm lunch meal!
- Changing experiences on how to use AST in detecting multidrug resistance microorganisms. In some lab, on market tablets are often used. These tablets are based mainly on synergy test

Dermatophyte

Dermatophyte
- Isolated from hair, nails
- Tricophyton, Microsporidia
- Macroscopic; flat, hairy, white to yellow
- Grow mainly in dermatohoyte specific medium (Sabouraud with added antibiotics and cyclohexmide to inhibit saprotrophic growth)
- Yeast more in nails of the finger, Aspegillus more in the nails of the toes

Yeast vs. mould
- Macroscopic: yeasts have shiny surface (look like m&ms candies)
- Yeast can grow within 1-3 days, moulds longer

Microscopic recognition
- Micro or macroconidia (conidia means spores)
- Mycellum is where the conidia hangs (spores can be free)
- Septae is the division of mycellum

Organizational stuff
- The specimens are placed in Sabouraud and dermatophyte specific medium
- When shiny surface colonies are seen: probably yeast
- When mould is suspected in dermatophyte specific medium --> reisolation in diluted Sabouraud, so spores are formed by the fungi (spores are formed in barren condition)
- When mould is suspected in usual Sabouraud --> reisolation in diluted Sabouraud and in dermatophyte specific medium

Tubes

Lithium heparine
- in The Netherlands: green cap, in Belgium: orange
- mainly for clinical chemistry: cholesterol, hormones, etc.

EDTA
- in The Netherlands: purple, in Belgium: rood
- EDTA maintains the cellular integrity
- so, it is used for the morphology of red blood cells, leukocytes

Serum
- in the Netherlands: red, in Belgium: white
- without anticoagulans, serum is retained
- so for serology

Citraat
- in the Netherlands: blue, in Belgium: green
- for coagulation

Thrombophilic tests

Pre-analysis
- patients should fullfilled screening criteria
- the tests should be performed before patients receive anticoagulant

Antithrombin deficiency
- blocks factor Xa and factor IIa (also factor IXa)
- the tests are made separately for factor Xa and IIa
- antithrombin activity tests:
  - antithrombin in the plasma of the patients binds with heparine in the reagens. Together they bind to FXa. The unbound FXa will be measured (it will bind with the substrates and measured).
 - the results are compared relative to the measurement of pooled plasma from normal patients.
 - in our lab <80% is considered as deficient.
 - remember that heparine cofactor can also bind to FIIa (not to FXa) and will lead to overesttimation of results.

- other tests are for example rocket immunoelectrophoresis that measure the antigen itself.

Protein C deficiency
- type I (low quantity), type 2 (low functionality)
- immunogenic, chromogenic techniques, and chromogenic with APTT

Protein S deficiency
- blocks factor VIIIa and Va
- assay is performed by measuring the activated protein C (the product), the activation is by the snake venom.

APC resisistency
- to measure factor V Leiden (the most common cause of non-challenged venous thrombosis)
- assay is by adding patient plasma with factor V deficient plasma. This mix is added with APTT reagens. The latter mix is added with APC and the other without APC. The ratio of the two is measured.