The potency of Tigecycline in the Sensititre plates
Michel [email protected]
Content• The Netherlands Food and
Consumer Product Safety Authority (NVWA)
• Antimicrobial susceptibilitytesting at the NVWA
• Tigecycline resistantEnterococci 2014
• Stability of Tigecycline
• Conclusion
The Netherlands Food and Consumer Product Safety Authority (NVWA)
The task and activities of the NVWA:To protect human and animal health by monitoring food and consumer products.
The Authority controls the whole production chain, from raw materials and processing aids to end products and consumption.3
Antimicrobial susceptibility testing 2014Strains: Panels Sensititre: Samples 2014 (n):Salmonella EUVSEC 291E. coli EUVSEC 1382 Enterococci EUVENC 1015Campylobacter EUCAMP2 402ESBL EUVSEC2 686
Equipment:
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Antimicrobial susceptibility testing NVWAThe results are annually reported to the EFSA and published in the MARAN(Monitoring of Antimicrobial Resistance and Antibiotic Usage in Animals in the Netherlands)
MARAN published by the CVI (Central Veterinary Institute) based on:• Total sales data and animal specific usage of antimicrobial agents in animal
husbandry.• Development of antimicrobial resistance in bacteria of animal origin.• Relevance to public health.
In collaboration with:
Netherlands Veterinary Medicines Authority NVWA RIVM
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Tigecycline resistant Enterococci 2014Introduction panel EUVENC January 2014
ANTIMICROBICSVAN VancomycinTEI TeicoplaninSYN Quinupristin /
dalfopristinTET TetracyclineDAP DaptomycinCIP CiprofloxacinERY ErythromycinTGC TigecyclineLZD LinezolidGEN GentamicinAMP AmpicillinCHL ChloramphenicolPOS Positive Control
*Resistant=>0.25 Tigecycline
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VAN128
TEI64
SYN64
TET128
DAP32
CIP16
ERY128
TGC4
LZD64
GEN1024
AMP64
CHL128
VAN64
TEI32
SYN32
TET64
DAP16
CIP8
ERY64
TGC2
LZD32
GEN512
AMP32
CHL64
VAN32
TEI16
SYN16
TET32
DAP8
CIP4
ERY32
TGC1
LZD16
GEN256
AMP16
CHL32
VAN16
TEI8
SYN8
TET16
DAP4
CIP2
ERY16
TGC0.5
LZD8
GEN128
AMP8
CHL16
VAN8
TEI4
SYN4
TET8
DAP2
CIP1
ERY8
TGC*0.25
LZD4
GEN64
AMP4
CHL8
VAN4
TEI2
SYN2
TET4
DAP1
CIP0.5
ERY4
TGC0.12
LZD2
GEN32
AMP2
CHL4
VAN2
TEI1
SYN1
TET2
DAP0.5
CIP0.25
ERY2
TGC0.06
LZD1
GEN16
AMP1
POSCON
VAN1
TEI0.5
SYN0.5
TET1
DAP0.25
CIP0.12
ERY1
TGC0.03
LZD0.5
GEN8
AMP0.5
POSCON
Tigecycline resistant Enterococci 2014 (n)
Enterococci examined: 1015Resistant for tigecycline (MIC>0.25): 263 (26%)
E. feacalis ATCC 29212: 31Resistant for tigecycline (MIC>0.25): 7 (23%)
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Batch EUVENCpanels used
Enterococci examined(n)
TIG Resistant (MIC>0.25)(n)
B3513 445 33B4255 458 165B4423 112 65
Total 1015 263
Stability of TigecyclineAntimicrob Agents Chemother. 2005 Sep; 49(9): 3903–3909. PMCID:PMC1195415Tigecycline MIC Testing by Broth Dilution Requires Use of Fresh Medium or Addition of the BiocatalyticOxygen-Reducing Reagent Oxyrase To Standardize the Test Method“It was previously established that tigecycline is susceptible to oxidative degradation in a solution”
PLoS One. 2014; 9(5): e95281.Published online 2014 May 28. doi:10.1371 PMCID:PMC4037176A Novel Formulation of Tigecycline Has Enhanced Stability and Sustained Antibacterial and AntileukemicActivity“In this study, we identified that the degradation of tigecycline after reconstitution is accelerated by exposure to light. It is well known that tigecycline degradation is oxygen-dependant”
J. of Chromatographic Sci. July 24, 2012. 192-9.doi: 10.1093 PMID:22832547Validation of a Stability-Indicating RP-LC Method for the Determination of Tigecycline in Lyophilized Powder“confirming the high reactivity of the tigecycline with oxygen, made possible by the low stability of this drug” “Under the photolytic condition, a significant decrease of the area was also observed”
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Stability of TigecyclineWe examined whether practical preparations could be the reason for these unexpected results.
Frequently, all EUVENC plates were stacked with the protective cover already removed for some time (in some cases 4 to 5 hours prior to analysis).
We suspected that this could explain theunexpected Tigecycline resistant isolates.
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Stability of Tigecycline
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SAMPLE NO. ORGANISM EUVENC panel results TGS NVWA 2014 EUVENC panel results TGC CVIpanel cover
removed 4 hours prior to analysispanel cover
removed just prior to analysisMIC (µg/ml) S / R MIC (µg/ml) S / R MIC (µg/ml) S / R
EF0464 E.FAEM = 0.06 S = 0.25 S = 0.06 SEF0612 E.FAEM = 0.06 S = 0.12 S = 0.06 SEF0846 E.FAEM = 0.06 S = 0.12 S = 0.06 SEF0906 E.FAEM = 0.06 S = 0.06 S = 0.06 SEF0914 E.FAEM = 0.06 S = 0.06 S = 0.06 SEF0482 E.FAEM = 0.12 S = 0.25 S = 0.06 SEF0605 E.FAEL = 0.12 S = 0.25 S = 0.12 SEF0611 E.FAEM = 0.12 S = 0.12 S = 0.12 SEF0616 E.FAEM = 0.12 S = 0.12 S = 0.12 SEF0646 E.FAEM = 0.12 S = 0.06 S = 0.06 SEF0753 E.FAEM = 0.12 S = 0.12 S = 0.06 SEF0863 E.FAEM = 0.12 S = 0.12 S = 0.06 SEF0898 E.FAEL = 0.12 S = 0.25 S = 0.12 SEF0902 E.FAEM = 0.12 S = 0.12 S = 0.12 SEF0442 E.FAEM = 0.25 S = 0.25 S = 0.25 SEF0527 E.FAEL = 0.25 S = 0.25 S = 0.12 SEF0619 E.FAEM = 0.25 S = 0.12 S = 0.06 SEF0633 E.FAEM = 0.25 S = 0.12 S = 0.06 SEF0835 E.FAEM = 0.25 S = 0.12 S = 0.06 SEF1443 E.FAEM = 0.25 S = 0.06 S = 0.06 SEF1446 E.FAEM = 0.25 S = 0.12 S = 0.06 SEF0592 E.FAEL = 0.5 R = 0.12 S = 0.12 SEF0821 E.FAEL = 0.5 R = 0.12 S = 0.06 SEF1501 E.FAEM = 0.5 R = 0.12 S = 0.12 SEF1502 E.FAEM = 0.5 R = 0.12 S = 0.06 S
Stability of Tigecycline
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SAMPLE NO. ORGANISM EUVENC panel results TGS NVWA 2014 EUVENC panel results TGC CVIpanel cover
removed 4 hours prior to analysispanel cover
removed just prior to analysisMIC (µg/ml) S / R MIC (µg/ml) S / R MIC (µg/ml) S / R
EF0464 E.FAEM = 1 R = 0.06 S = 0.12 SEF0612 E.FAEM = 1 R = 0.06 S = 0.06 SEF0846 E.FAEM = 1 R = 0.25 S = 0.12 SEF0906 E.FAEM = 1 R = 0.06 S = 0.06 SEF0914 E.FAEM = 1 R = 0.25 S = 0.06 SEF0482 E.FAEM = 1 R = 0.12 S = 0.06 SEF0605 E.FAEL = 1 R = 0.12 S = 0.25 SEF0611 E.FAEM = 1 R = 0.12 S = 0.06 SEF0616 E.FAEM = 1 R = 0.12 S = 0.12 SEF0646 E.FAEM = 1 R = 0.12 S = 0.12 SEF0753 E.FAEM = 1 R = 0.25 S = 0.12 SEF0863 E.FAEM = 1 R = 0.06 S = 0.06 SEF0898 E.FAEL = 1 R = 0.12 S = 0.25 SEF0902 E.FAEM = 1 R = 0.12 S = 0.25 SEF0442 E.FAEM = 1 R = 0.12 S = 0.12 SEF0527 E.FAEL = 1 R = 0.12 S = 0.12 SEF0619 E.FAEM = 1 R = 0.12 S = 0.25 SEF0633 E.FAEM = 1 R = 0.12 S = 0.25 SEF0835 E.FAEM = 1 R = 0.25 S = 0.12 SEF1443 E.FAEM = 1 R = 0.25 S = 0.12 SEF1446 E.FAEM = 1 R = 0.12 S = 0.12 SEF0592 E.FAEL = 1 R = 0.12 S = 0.12 SEF0821 E.FAEL = 1 R = 0.12 S = 0.12 SEF1501 E.FAEM = 1 R = 0.12 S = 0.12 SEF1502 E.FAEM = 1 R = 0.12 S = 0.06 S
Stability of Tigecycline
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ConclusionThese results suggest (in our case) that:
• The prolonged exposure of Tigecycline to light and air caused photo-oxidation and thus Tigecycline degradation.
• After removing the panel cover prior to analysis, and keeping the panel prior to incubation in the dark, no more Tigecycline resistant Enterococci were identified.
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Advice
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Stay on the darkside with your panels
ACKNOWLEDGEMENT NVWA:Heymans, dr. R. (Raymond)Wit, ing. B. (Ben)Nap, I.D. (Irma)Nabuurs, ing. F.C.P. (Fiona)Sijben, F.H.G. (Frank)Nendos, N. (Nico)Engelen, R.J.K. van (Ruud)Smit, C. (Corry)
CVI:Veldman, dr. T. (Kees)15
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