Espinosa-Gongora, Carmen

Hansen, Julie Elvekjær

Larsen, A. R.

Larsen, J.

Pedersen, Karl

Skov, R.

Sørensen, Anna Irene Vedel

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Downloaded from orbit.dtu.dk on: Dec 21, 2017Assessment of Methods to Quantify Livestock Associated MRSA in Pig HerdsHansen, Julie Elvekjær; Sørensen, Anna Irene Vedel; Espinosa-Gongora, Carmen; Larsen, A. R.; Larsen,J.; Skov, R.; Pedersen, KarlPublication date:2015Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Hansen, J. E., Sørensen, A. I. V., Espinosa-Gongora, C., Larsen, A. R., Larsen, J., Skov, R., & Pedersen, K.(2015). Assessment of Methods to Quantify Livestock Associated MRSA in Pig Herds. Poster session presentedat 4th ASM-ESCMID Conference on Methicillin-resistant Staphylococci in Animals: Veterinary and Public HealthImplications, Chicago, Illinois, United States.Methods to quantify livestock associated MRSA in pig herds Julie Elvekjær Hansen1, Anna Irene Vedel Sørensen1, Carmen Espinosa-Gongora2, Anders Rhod Larsen3, Jesper Larsen3, Robert Skov3 and Karl Pedersen1 1Technical University of Denmark, National Veterinary Institute, Frederiksberg, Denmark, 2University of Copenhagen, Department of Veterinary Disease Biology, Frederiksberg, Denmark,3Statens Serum Institut, Copenhagen, Denmark.  DTU Vet National Veterinary Institute Section of Bacteriology and Pathology  Bülowsvej 27, Frederiksberg Denmark Fig. 1. E.g. of plates from dilution serie of an animal sample 1A) 100 1B) 10-1 1C) 10-2 and from a 250L sampling with Sampl’air in duplicates on selective plates, 1D & 1E, and on blood agar 1F. 1A 1B 1C 1D 1E 1F Background A high prevalence of LA-MRSA, especially in the pig production, has been reported from many countries. However, there is a lack of knowledge about the actual LA-MRSA loads on infected farms. The overall MRSA level is low in Denmark and if the reservoir of LA-MRSA starts to spread into the community, the MRSA level in general could increase. We do not believe it is possible to eliminate LA-MRSA in the pig production, but we have an ambition of being able to lower the level. In order to try and control LA-MRSA, it is highly relevant to be able to quantify LA-MRSA in the farm environment and on livestock which would enable us to rank risk factors in the production chain and to assess the effect of possible intervention strategies.  Conclusion Fast quantification of the animal MRSA load in pig herds is possible by direct plating of either nasal or skin swab samples. The Sartorius MD8 air sampler provides fast environmental quantification and it was possible to detect MRSA in flies with animal contact demonstrating their potential as environmental MRSA carriers.  Purpose The present study was undertaken to assess methods to quantify LA-MRSA on animals and in the environment in pig herds. In addition, the role of flies as potential environmental carriers of LA-MRSA was investigated. An example of plates from direct plating and air sampling is seen in figure 1. juhan@vet.dtu.dk vet.dtu.dk Air samples on MRSA 2 Direct plating of swabs on MRSA 2 Colony count Cfu determination Method Range of MRSA load Nose 6.6-×101-3.9×104 cfu/swab Skin 1.1×101-2.6×105 cfu/swab MD8 4.1×102-6.3×102 cfu/m3 Table 2. Range of MRSA loads. Source Nasal swabs Skin swabs   Variance % Variance % Bio. replicate 0.37 24 0.08 31 Pig 1.11 72 0.13 53 Residual 0.06 4 0.04 15 Total 1.54 100 0.25 100 CFU based on counts from 10-0, where the 10-1 counts were zero Table 1. Proportions of variance Nasal swabs seemed to cause greater variation in the results than use of skin swabs and a larger proportion of the variance could be explained by the difference between pigs.  The Sartorius air sampler yielded the most stable detection level of LA- MRSA and the increasing raw counts detected corresponded better with increasing air volumes. Loads in the 15 weaning pigs supported the results seen in the method verification. Air load and skin load seemed similar while nasal load was higher. MRSA was found on flies, in pools of 5 and individually, after enrichment.   Flies Individual Pool of 5 Farrowing sectiona - 1/1 Weaning sectiona - 1/1 Farrowing sectionb 0/5 0/2 Weaning sectionb 4/5 2/2 a) Animal activity in both sections b) Animal activity only in weaning section Method verification Results Airborne LA-MRSA  Load in weaning pigs (n=15) 1 2 pig 3 Sample volume in liters Nasal and skin swabs per weaner Log(CFU/swab) Log(CFU/swab) CFU/m3 Air sampler • Sample’air • Sartorius (MD8)  Type • Nasal • Skin Type • Nasal • Skin Avg. raw count Sample volume in liters Air sampler • Sample’air • Sartorius (MD8)  

Assessment of Methods to Quantify Livestock Associated MRSA in Pig Herds

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Assessment of Methods to Quantify Livestock Associated MRSA in Pig Herds

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