AHEAD IN ANIMAL HEALTH
AST results of S. Dublin and S. Typhimurium from veal calves in the Netherlands, 2016 - 2020 Annet Heuvelink1*, Jobke van Hout1, Thijs Derkman1, Marieke Augustijn1, Els Broens2, Maaike Gonggrijp1 1 Royal GD, Deventer, the Netherlands, 2Utrecht University, Utrecht, the Netherlands, * Presenting author
Aim of the study
Conclusions
Assessing the representativeness and reliability of antimicrobial sensitivity testing (AST) results available for Salmonella Dublin (SDU) and Salmonella Typhimurium (STY) from clinical samples from Dutch veal calves.
The low number of isolates does lead to less representative and reliable AST results. Therefore, effort should be made to increase the number of isolates from veal calves.
Methods AST results (Minimal Inhibitory Concentrations (MICs)) and additional data (e.g., location of the farm and the associated veterinary practice) were extracted from the Laboratory Information Management System of Royal GD, from 2016 - 2020. After validation of the data, multilevel mixed-effects logistic regressions were carried out (STATA 15.1) to study associations between AST results and different parameters.
Results Low numbers of isolates • After validation of the data, 202 unique SDU and 316 STY isolates remained available for analyses (Table 1). • Because of the relatively low numbers of SDU and STY, the determined resistance percentages were less precise, especially when results of only one year were presented, with 95% confidence intervals ranging up to 30%. • The number of STY isolates even decreased during the study period.
Table 1: Number of isolates per year per serovar. Serovar Salmonella Dublin Salmonella Typhimurium
2016 2017 2018 2019 2020 42 86
45 75
39 78
42 44
34 33
Positive trend antimicrobial susceptibility • The MIC distributions for SDU and STY are shown in Table 2. • Results showed a positive trend for antimicrobial susceptibility of SDU and STY from 2016 to 2020, with decreasing odds for resistance for the majority of included antimicrobials. Table 2: MIC distribution for Salmonella Dublin and Salmonella Typhimurium isolates from veal calves, years 2016 - 2020. Salmonella Dublin 2016 − 2020 (n=202) MIC values (µg/mL)
Antimicrobial Ampicillin Apramycin Cefepime Cefotaxime Colistin Enrofloxacin Florfenicol Flumequine Gentamicin Neomycin Streptomycin Tetracycline Trim/Sulfaa
0.25
0.5
1
2
4
8
16
32
64
3.5
51.0
21.8
4.0
0.5 0.0 0.0 73.3 0.0 54.0 0.5 1.0 93.6 28.2 0.0 0.0
0.0 0.0 0.0
1.0 0.5
0.5 0.0 10.9 0.0 23.3 95.5 83.7
0.0 0.0 0.0
19.3
99.5 100.0 13.9 0.0
0.0 100.0 0.0 0.0 0.5
0.0
0.5
99.5
0.5 79.2
8.4 1.0
62.9 0.0
15.8 6.4 0.0
3.5 2.0 1.5 0.0 31.2 0.0 19.8
19.3 2.0 13.9 0.5 4.5 0.0
128
0.0
0.0 5.9 1.5 21.8
0.0
18.8
Salmonella Typhimurium 2016 − 2020 (n=316) MIC values (µg/mL)
MIC50
MIC90
S
R
R CI
(μg/ mL)
(μg/ mL)
(%)
(%)
(%)
0.25
0.5
1
2
4
8
16
32
64
0.5 ≤8 ≤1 ≤1 4 ≤0.25 4 ≤2 ≤2 ≤4 8 1 ≤0.25
>32 ≤8 ≤1 ≤1 4 ≤0.25 >8 ≤2 >8 ≤4 >64 >16 >4
80.7 99.5 100.0 99.5 95.5 84.7 93.6 79.7 78.2 80.2
19.3 0.5 0.0 0.5 4.5 15.3 6.4 20.3 21.8 19.8
14.1-25.4 0.0-2.7 0.0-1.8 0.0-2.7 2.1-8.3 10.7-21.1 3.5-10.8 15.0-26.5 16.3-28.1 14.5-26.0
0.0
5.1
41.1
7.0
0.6 0.0 0.0 0.3 0.0 54.1 0.3 0.3 94.9 2.8 0.6 0.0
0.3 0.0 0.0
76.6 0.6
0.0 0.3 2.5 0.9 1.9 96.8 67.4
0.0 0.3 0.0
45.9
100.0 99.7 20.3 0.6
0.0 99.7 0.0 0.0 0.0
0.0
0.3
2.5 0.9 3.2 0.3 3.2 0.0 13.6
41.5 1.6 29.1 0.3 2.2 1.9
97.8
0.0 80.7
0.0 5.7
3.5 0.0
1.6 1.9 0.0
128
0.0
0.3 4.4 5.7 92.1
27.5
57.0
MIC50
MIC90
S
R
R CI
(μg/mL)
(μg/mL)
(%)
(%)
(%)
2 ≤8 ≤1 ≤1 ≤0.5 ≤0.25 4 ≤2 ≤2 ≤4 >64 >16 ≤0.25
>32 ≤8 ≤1 ≤1 1 ≤0.25 >8 ≤2 >8 ≤4 >64 >16 >4
53.8 100.0 99.7 96.8 67.7 94.9 9.8 6.0 86.4
46.2 0.0 0.3 3.2 32.3 5.1 90.2 94.0 13.6
40.6-51.9 0.0-1.2 0.0-1.8 1.5-5.7 27.2-37.7 2.9-8.1 86.4-93.2 90.8-96.3 10.0-17.9
Dilution series applied for each individual antimicrobial are marked white, green vertical lines indicate the breakpoint/cut off used for interpretation. To the right of the dilution ranges, percentages of isolates with a MIC value higher than the highest concentration of the dilution range are mentioned in red. The percentage of isolates mentioned at the lowest concentration of a dilution range, refers to isolates with a MIC value equal to or lower than the lowest concentration evaluated in the specific dilution range. a Only the concentration of trimethoprim, tested in a 1:19 ratio (trimethoprim:sulfamethoxazole) is mentioned. n = number of observations; MIC50 = Minimum Inhibitory Concentration required to inhibit the growth of 50% of the isolates; MIC90 = Minimum Inhibitory Concentration required to inhibit the growth of 90% of the isolates; S = susceptible; R = resistant; CI = 95% Confidence Interval.
Representativeness • The distribution of SDU and STY isolates across the Netherlands was similar to the population density of veal calves (Figure 1). • The individual provinces did not have significant different odds for isolates being resistant. • Isolates originating from farms associated with the same veterinary practice were more alike than isolates originating from farms associated with different veterinary practices.
Figure 1. Distribution of the farms of origin of SDU and STU isolates in the dataset of 2016 - 2020.
Funding by The Netherlands Organisation for Health Research and Development (ZonMw); Dossier number: 541003007.
a.heuvelink@gdanimalhealth.com www.gdanimalhealth.com
GD2825/03-23