Canine pyoderma: mecA persists autogenous bacterin formulation from meticillin-resistant Staphylococcus pseudintermedius (MRSP) and S. aureus (MRSA)

Abstract

Objective Autogenous Staphylococcus pseudintermedius bacterins can reduce prescribing of antimicrobials in the management of canine recurrent pyoderma. However, increasing prevalence of meticillin-resistant, mecA-positive S. pseudintermedius (MRSP) raises concern over dispersal of mecA through bacterin therapy. We investigated the presence and integrity of mecA in bacterin formulations after manufacturing.

Material and methods Twenty clinical isolates (12 MRSP, 7 MR-S. aureus, 1 meticillin-susceptible SP) were investigated. Pellets from overnight growth were washed 3 times with 0.5 % phenol saline, followed by addition of 0.1 ml 10 % formal-saline to 10 ml phenol-saline. Sterility was confirmed, and DNA extracted using both a standard genomic extraction kit and one recommended for formalin-fixed tissue samples (FFPE). The presence of mecA was determined after PCR and its integrity examined in 5 randomly selected samples after sequencing.

Results In all bacterins from meticillin-resistant isolates, mecA was detected following FFPE extraction; products aligned fully to a reported mecA sequence. After standard DNA extraction, mecA was seen in 16/19 samples.

Conclusion Persistence of mecA in MRSP bacterins suggests that dispersal of this important resistance mediator through therapy may be possible. While the ability of skin bacteria to uptake naked DNA remains unclear, it seems prudent to only formulate autogenous bacterins from mecA-negative S. pseudintermedius to avoid unnecessary spread of mecA.

Zusammenfassung

Ziel der Studie Autogene Staphylococcus pseudintermedius-Bakterine (Autovakzinen) können helfen, die Verschreibung von Antibiotika in der Therapie rezidivierender Pyodermien des Hundes zu reduzieren. Die Zunahme von Infektionen mit Meticillin-resistenten, mecA-positiven Staphylococcus pseudintermedius (MRSP) ist besorgniserregend, da eine Bakterintherapie möglicherweise zur Verbreitung von mecA beitragen kann. Wir untersuchten daher das Auftreten und die Integrität von mecA in Autovakzinen nach der Herstellung.

Material und Methoden Zwanzig klinische Isolate (12 MRSP, 7 MR-S. aureus, 1 Meticillin-empfindlicher SP) wurden untersucht. Zellpellets wurden 3-mal mit 0,5 %iger Phenol-Kochsalzlösung gewaschen, danach folgte eine Zugabe von 0,1 ml 10 %iger Formalin-Kochsalzlösung zu 10 ml Phenol-Kochsalzlösung. Sterilität wurde bestätigt und DNA mit einem standardisierten genomischen Extraktionskit sowie einem weiteren für formalinfixiertes Gewebe (FFPE) extrahiert. Die Präsenz von mecA wurde mittels PCR bestimmt und seine Integrität in 5 zufällig ausgewählten Proben nach Sequenzierung untersucht.

Ergebnisse In allen Bakterinen aus Meticillin-resistenten Isolaten ließ sich mecA nach FFPE-Extraktion nachweisen. Die Produkte wurden alle mit einem beschriebenen mecA abgeglichen. Nach einer Standard-DNA-Extraktion fand sich mecA in 16/19 Proben.

Klinische Relevanz Das Verbleiben von mecA in MRSP-Bakterinen weist auf die Möglichkeit einer Verbreitung dieses wichtigen Resistenzmediators durch eine Bakterintherapie hin. Solange die Fähigkeit von Hautbakterien, nackte DNA aufzunehmen, unklar ist, sollten autogene Bakterine nur aus mecA-negativen S. pseudintermedius hergestellt werden, um das Risiko einer Verbreitung von mecA zu vermeiden.

Key words

Antimicrobial resistance - superficial pyoderma - recurrent - phenol - formalin - DNA

Schlüsselwörter

Antibiotikaresistenz - superfizielle Pyodermie - rezidivierend - Phenol - Formalin - DNA

Publikationsverlauf

Eingereicht: 05. November 2020

Angenommen: 18. Mai 2021

Artikel online veröffentlicht:
03. Dezember 2021

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