Molecular characterization of the <i>HMTp210</i> gene of <i>Avibacterium paragallinarum</i> and the proposition of a new genotyping method as alternative for classical serotyping

Buter, Rianne; Feberwee, Anneke; Wit, Sjaak de; Heuvelink, Annet; Silva, Ana da; Gallardo, Rodrigo A.; Soriano‐Vargas, Edgardo; Swanepoel, Stefan; Jung, Arne; Todte, Matthias; Dijkman, Remco

doi:10.1080/03079457.2023.2239178

Cited by 2 publications

(2 citation statements)

References 29 publications

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“…The PCR method described above was used to test all isolates, and an optimized PCR program was used. The results were contrasted with that of the hmtp210 sequence analysis method ( Buter et al, 2023 ).…”

Section: Methodsmentioning

confidence: 99%

“…The mPCR exhibited limited efficacy in identifying isolates of A. paragallinarum that are classified under page serovar C. Recent studies have demonstrated that genetic evolutionary analyses utilizing partial sequences of the hmtp210 gene can be used to differentiate between serotypes A, B, and C ( Tan et al, 2020 ; Tan et al, 2021 ). Analysis of the hmtp210 hypervariable region alone did not provide enough discriminatory power to distinguish different Kume serovar strains ( Buter et al, 2023 ). Thus, the hmtp210 gene is closely associated with the serotyping of A. paragallinarum .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Prevalence and genomic-based antimicrobial resistance analysis of Avibacterium paragallinarum isolates in Guangdong Province, China

Cao,

Huang,

Lin

et al. 2024

Poultry Science

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Prevalence and genomic-based antimicrobial resistance analysis of Avibacterium paragallinarum isolates in Guangdong Province, China

Cao,

Huang,

Lin

et al. 2024

Poultry Science

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Recommendation of a standardized broth microdilution method for antimicrobial susceptibility testing of Avibacterium paragallinarum and resistance monitoring

Gütgemann,

Heuvelink,

Müller

et al. 2024

J Clin Microbiol

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This study aimed to develop a method for standardized broth microdilution antimicrobial susceptibility testing (AST) of Avibacterium ( Av .) paragallinarum , the causative agent of infectious coryza in chickens. For this, a total of 83 Av. paragallinarum isolates and strains were collected from 15 countries. To select unrelated isolates for method validation steps, macrorestriction analyses were performed with 15 Av. paragallinarum . The visible growth of Av. paragallinarum was examined in six broth media and growth curves were compiled. In Veterinary Fastidious Medium and cation-adjusted Mueller-Hinton broth (CAMHB) + 1% chicken serum + 0.0025% NADH (CAMHB + CS + NADH), visible growth of all isolates was detected and both media allowed adequate bacterial growth. Due to the better readability of Av. paragallinarum growth in microtiter plates, CAMHB + CS + NADH was chosen for AST. Repetitions of MIC testing with five epidemiologically unrelated isolates using a panel of 24 antimicrobial agents resulted in high essential MIC agreements of 96%–100% after 48-h incubation at 35 ± 2°C. Hence, the remaining 78 Av. paragallinarum were tested and demonstrated easily readable MICs with the proposed method. Differences in MICs were detected between isolates from different continents, with isolates from Africa showing lower MICs compared to isolates from America and Europe, which more often showed elevated MICs of aminoglycosides, quinolones, tetracyclines, and/or trimethoprim/sulfamethoxazole. PCR analyses of isolates used for method development revealed that isolates with elevated MICs of tetracyclines harbored the tetracycline resistance gene tet (B) but none of the other tested resistance genes were detected. Therefore, whole-genome sequencing data from 62 Av. paragallinarum were analyzed and revealed the presence of sequences showing nucleotide sequence identity to the genes aph(6)-Id , aph(3″)-Ib , bla TEM-1B , catA2, sul2 , tet (B), tet (H), and mcr- like. Overall, the proposed method using CAMHB + CS + NADH for susceptibility testing with 48-h incubation time at 35 ± 2°C in ambient air was shown to be suitable for Av. paragallinarum . Due to a variety of resistance genes detected, the development of clinical breakpoints is highly recommended. IMPORTANCE Avibacterium paragallinarum is an important pathogen in veterinary medicine that causes infectious coryza in chickens. Since antibiotics are often used for treatment and resistance of the pathogen is known, targeted therapy should be given after resistance testing of the pathogen. Unfortunately, there is currently no accepted method in standards that allows susceptibility testing of this fastidious pathogen. Therefore, we have worked out a method that allows harmonized susceptibility testing of the pathogen. The method meets the requirements of the CLSI and could be used by diagnostic laboratories.

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Molecular characterization of the HMTp210 gene of Avibacterium paragallinarum and the proposition of a new genotyping method as alternative for classical serotyping

Cited by 2 publications

References 29 publications

Prevalence and genomic-based antimicrobial resistance analysis of Avibacterium paragallinarum isolates in Guangdong Province, China

Prevalence and genomic-based antimicrobial resistance analysis of Avibacterium paragallinarum isolates in Guangdong Province, China

Recommendation of a standardized broth microdilution method for antimicrobial susceptibility testing of Avibacterium paragallinarum and resistance monitoring

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