Isolation and molecular identification of Avibacterium paragallinarum in suspected cases of infectious coryza

Badouei, Mahdi Askari; SADRZADEH, Avesta; AZAD, Nader; Blackall, P. J.; MADADGAR, Omid; CHARKHKAR, Saeid

doi:10.3906/vet-1301-62

Cited by 9 publications

(24 citation statements)

References 8 publications

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“…All isolates were V‐factor dependent, showing typical ‘satellite’ growth on solid plate cultures. Consistent with other studies (Ahmadi & Nasri, 2018; Badouei et al., 2014; Banani et al., 2007; Blackall & Soriano‐Vargas, 2020; Blackall et al., 1990; Falconi‐Agapito et al., 2015; Gallardo et al., 2020; Han et al., 2016; Luna‐Galaz et al., 2016; Nouri et al., 2020; Vargas & Terzolo, 2004), the result of the biochemical identification method was the isolation of Gram‐negative, catalase‐negative, oxidase‐negative coccobacilli, which were d‐xylose and trehalose‐negative in fermentation, but were able to ferment d ‐mannitol and maltose. These are some of the biochemical features of bacteria that cause IC (Blackall & Soriano‐Vargas, 2020).…”

Section: Discussionsupporting

confidence: 93%

Retracted: Novel NAD‐independent Avibacterium paragallinarum: Isolation, characterization and molecular identification in Iran

Beiranvand

Piri-Gharaghie

Dehganzad

et al. 2022

Veterinary Medicine & Sci

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Background: Infectious coryza (IC) is an invasive upper respiratory disease caused by Avibacterium paragallinarum that affects birds, particularly chickens. The objective of this study is to isolate, characterize and molecularly identify the bacterium A. paragallinarum in poultry birds, as well as to determine its antibiotic sensitivity and resistance.Methods: A total of 10 chickens from four different Iranian farms with typical IC symptoms were used in this study. The nasal swabs were streaked onto chocolate agar plates and blood agar plates and incubated at 37 • C in 5% CO 2 for 24 to 48 h. As part of the identification of bacteria, bacteriological observations and polymerase chain reaction (PCR) testing are conducted. The antibiotic sensitivity tests were also performed using the disk diffusion method against A. paragallinarum and the prevalence in different farms was determined.Results: By using biochemical assays and PCR analyses, seven strains of A. paragallinarum were isolated from samples of four chicken farms with typical IC clinical signs.Most isolates (4/7) showed the typical requirement for nicotinamide adenine dinucleotide (NAD) and an enriched CO 2 atmosphere for growth. Three of the seven strains of A. paragallinarum were found to be novel NAD-independent under anaerobic conditions. There was one biochemical biovar identified in terms of carbohydrate fermentation patterns, although changes in maltose carbohydrate fermentation patterns were detected in the No. 5 strain. All isolates were sensitive to gentamicin and spectinomycin. Three novel NAD-independent strains (Nos.1, 5 and 7) were found to be multidrug-resistant (MDR) and resistant to at least three classes of antibiotics. There was greater antibiotic resistance in the three NAD-independent isolates than in normal NAD-dependent bacteria. Conclusion:By discovering NAD-independent forms of A. paragallinarum, these species have a greater range than previously believed. A clear, cautious approach should be taken in diagnosing and possibly controlling IC.

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Section: Discussionsupporting

confidence: 93%

Retracted: Novel NAD‐independent Avibacterium paragallinarum: Isolation, characterization and molecular identification in Iran

Beiranvand

Piri-Gharaghie

Dehganzad

et al. 2022

Veterinary Medicine & Sci

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“…2,6,10 Subclinical carrier birds can be a reservoir for transmission via direct contact, airborne droplets, or fomites. 1,11 Concurrent respiratory agents, including Mycoplasma synoviae (MS), M. gallisepticum (MG), Pasteurella sp., and infectious bronchitis virus (IBV; species Avian coronavirus), as well as stress factors, can exacerbate disease. 1,9 A. paragallinarum is a fastidious gram-negative bacterium in the Pasteurellaceae family.…”

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confidence: 99%

“…1,11 Concurrent respiratory agents, including Mycoplasma synoviae (MS), M. gallisepticum (MG), Pasteurella sp., and infectious bronchitis virus (IBV; species Avian coronavirus), as well as stress factors, can exacerbate disease. 1,9 A. paragallinarum is a fastidious gram-negative bacterium in the Pasteurellaceae family. 3,5 Detection of A. paragallinarum by culture is often hindered both by its fastidious characteristics (slow growth rate, need for specialized media and growth conditions, and minimal reactivity to standard biochemical tests) and the presence of other bacterial organisms present in sample sites, including Gallibacterium anatis, Pasteurella multocida, and Escherichia coli, among others.…”

mentioning

confidence: 99%

“…3,5 Detection of A. paragallinarum by culture is often hindered both by its fastidious characteristics (slow growth rate, need for specialized media and growth conditions, and minimal reactivity to standard biochemical tests) and the presence of other bacterial organisms present in sample sites, including Gallibacterium anatis, Pasteurella multocida, and Escherichia coli, among others. 1,5 Delays in sample processing and previous antimicrobial treatment have been shown to prevent recovery. 2 Vaccination, which is common in affected commercial operations, conveys strong protection against homologous strains of A. paragallinarum.…”

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confidence: 99%

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Surveillance for Avibacterium paragallinarum in autopsy cases of birds from small chicken flocks using a real-time PCR assay

2019

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Infectious coryza is a severe respiratory disease of chickens associated with large economic losses in affected commercial flocks. The fastidious causative pathogen, Avibacterium paragallinarum, is difficult to recover and identify, resulting in delayed diagnosis and enhanced spread of the agent. Small poultry flocks are increasingly common in rural and suburban environments. We assessed the frequency of A. paragallinarum using real-time PCR and clinical conditions present in samples from such flocks submitted to the California Animal Health and Food Safety Laboratory System (Davis, CA) in 2018. From the 294 samples collected for our study, 86 (30%) were PCR-positive for A. paragallinarum. Juvenile birds (≤1 y) were significantly more likely to be PCR-positive (p = 0.017), and birds diagnosed with respiratory disease had lower Ct values (p = 0.001) than those without. Concurrent infections were also identified, including with Mycoplasma gallisepticum (18.6%), M. synoviae (18.6%), infectious bronchitis virus (12.8%), and infectious laryngotracheitis virus (7.0%). Only 46.5% of PCR-positive chickens had antemortem respiratory signs, making endemic infections in these flocks highly likely. Our study demonstrates that A. paragallinarum is present in small-flock operations including those without respiratory disease and may present a risk for airborne pathogen transmission to commercial poultry operations.

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Validation of a real-time PCR assay for high-throughput detection of Avibacterium paragallinarum in chicken respiratory sites

et al. 2019

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Avibacterium paragallinarum is the causative agent of infectious coryza, a highly contagious respiratory disease in chickens. Given its fastidious nature, this bacterium is difficult to recover and identify, particularly from locations colonized by normal bacterial flora. Standard PCR methods have been utilized for detection but are labor-intensive and not feasible for high-throughput testing. We evaluated a real-time PCR (rtPCR) method targeting the HPG-2 region of A. paragallinarum, and validated a high-throughput extraction for this assay. Using single-tube extraction, the rtPCR detected 4 A. paragallinarum (ATCC 29545 T and 3 clinical) isolates with a limit of detection (LOD) of 10 cfu/mL and a PCR efficiency of 89-111%. Cross-reaction was not detected with 33 non-A. paragallinarum, all close relatives from the family Pasteurellaceae. Realtime PCR testing on extracts of 66 clinical samples (choana, sinus, or trachea) yielded 98.2% (35 of 36 on positives, 30 of 30 on negatives) agreement with conventional PCR. Duplicate samples tested in a 96-well format extraction in parallel with the single-tube method produced equivalent LOD on all A. paragallinarum isolates, and 96.8% agreement on 93 additional clinical samples extracted with both procedures. This A. paragallinarum rtPCR can be utilized for outbreak investigations and routine monitoring of susceptible flocks.

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Isolation and molecular identification of Avibacterium paragallinarum in suspected cases of infectious coryza

Cited by 9 publications

References 8 publications

Retracted: Novel NAD‐independent Avibacterium paragallinarum: Isolation, characterization and molecular identification in Iran

Retracted: Novel NAD‐independent Avibacterium paragallinarum: Isolation, characterization and molecular identification in Iran

Surveillance for Avibacterium paragallinarum in autopsy cases of birds from small chicken flocks using a real-time PCR assay

Validation of a real-time PCR assay for high-throughput detection of Avibacterium paragallinarum in chicken respiratory sites

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