The prevention of avian colibacillosis has historically been investigated through vaccination, with variable outcomes. Commercial live (attenuated) and inactivated vaccines are reported to have limited efficacy in the context of heterologous challenge. Autogenous vaccination, using field isolates, is widely used, but scarcely documented. Different vaccination programs, including a live commercial vaccine and/or an inactivated autogenous vaccine, were compared for three different avian pathogenic Escherichia coli (APEC) strain (serotypes O78, O18 and O111) challenges. On the pullet farm, four groups of conventional pullets received different vaccination protocols. Group A was kept unvaccinated (control group). Group B was vaccinated three times with a live commercial O78 E. coli vaccine (at one day old, 59 and 110 days of age). Group C was immunized twice (at 79 and 110 days) with a three-valence autogenous vaccine (O78, O18 and O111). Group D was vaccinated first with the commercial vaccine (at one day old and 59 days), then with the autogenous vaccine (110 days). Birds were transferred to the experimental facility at 121 days of age and were challenged 10 days later. In each group, 20 birds were challenged with one of the three APEC strains (O78, O18, O111); in total, 80 birds were challenged by the same strains (20 per group). The recorded outcomes were: mortality rate, macroscopic lesion score in target organs and the bacterial recovery of the challenge strain from bone marrow and pooled organs. When challenged with O78 or O111 strains, birds from groups C and D proved to be significantly better protected, in terms of lesion scoring and bacteriological isolation, than those of groups A and B. With the O18 challenge, only birds of group D presented a statistically significant reduction of their lesion score. To the authors’ knowledge, this is the first report on the efficacy of an immunization program in poultry that combines commercial and autogenous vaccines.

Aeromonas salmonicida subsp. salmonicida is a pathogenic bacterium responsible for furunculosis in salmonids. Following an outbreak of furunculosis, the infection can be treated with antibiotics, but it is common to observe ineffective treatment due to antibiotic resistance. This bacterium has a wide variety of plasmids responsible for this resistance. Among them, pRAS3 carries a tetracycline resistance gene. Several variants of this plasmid have been discovered over the years (pRAS3-3432 and pRAS3.1 to 3.4). During the present study, two new variants of the plasmid pRAS3 were identified (pRAS3.5 and pRAS3-3759) in strains of A. salmonicida subsp. salmonicida. Plasmid pRAS3-3759, which has been found in many strains from the same region over the past three years, has an additional genetic element identical to one found in pRAS3-3432. This genetic element was also found in Chlamydia suis, a swine pathogen. In this study, we analyzed the bacteria’s resistance to tetracycline, the number of copies of the plasmids, and the growth of the strains that carry five of the pRAS3 variants (pRAS3.3 to 3.5, pRAS3-3432, and pRAS3-3759). The results show no particular trend despite the differences between the plasmids, except for the resistance to tetracycline when analyzed in an isogenic background. Blast analysis also revealed the presence of pRAS3 plasmids in other bacterial species, which suggests that this plasmid family has widely spread. This study once again highlights the ability of A. salmonicida subsp. salmonicida to adapt to furunculosis antibiotic treatments, and the still-growing family of pRAS3 plasmids.

Actinobacillus pleuropneumoniae is a major swine pathogen, classified into 19 serotypes based on capsular polysaccharide (CPS) loci. This study aimed to improve the diagnostic method to differentiate between serotypes 9 and 11, which are challenging to distinguish using conventional serological and molecular methods. A novel qPCR assay based on locked nucleic acid (LNA) probes was developed and validated using a collection of reference strains representing all known 19 serotypes. The assay demonstrated specificity in detecting the nucleotide variation characteristic of the serotype 9 reference strain. However, the analysis of a clinical isolate collection identified discrepancies between LNA-qPCR and serological results, prompting further investigation of the cps and O-Ag loci. Subsequent nanopore sequencing and whole-genome sequencing of a collection of 31 European clinical isolates, previously identified as serotype 9, 11, or undifferentiated 9/11, revealed significant genetic variations in the cps and O-Ag loci. Ten isolates had a cpsF sequence identical to that of the serotype 11 reference strain, while six isolates had single-nucleotide polymorphisms that were unlikely to cause significant coding changes. In contrast, 15 isolates had interruptions in the cpsF gene, distinct from that found in the serotype 9 reference strain, potentially leading to a serotype 9 CPS structure. In the O-Ag loci, differences between serotypes 9 and 11 were minimal, although some isolates had mutations potentially affecting O-Ag expression. Overall, these findings suggest that multiple genetic events can lead to the formation of a serotype 9 CPS structure, hindering the development of a single qPCR assay capable of detecting all cpsF gene mutations. Our results suggest that, currently, a comprehensive analysis of the cpsF gene is necessary to accurately determine whether the capsule of an isolate corresponds to serotype 9 or 11. Although such analyses are feasible with the advent of third-generation sequencing technologies, their accessibility, cost, and time to result limit their use in routine diagnostic applications. Under these circumstances, the designation of the hybrid serovar 9/11 remains a valid approach.

Background/Objectives: Pasteurella multocida is responsible for a wide variety of animal diseases worldwide, causing major economic losses. These infections are usually treated with antibiotics; however, the emergence of multidrug-resistant (MDR) strains is increasingly hindering. Understanding antibiotic resistance in P. multocida is important for effective treatment strategies and public health, as it impacts both animal and human welfare. Methods: In this study, the antibiotic susceptibility of 80 P. multocida isolates was evaluated by phenotypic (disk diffusion and broth microdilution) and genotypic analysis via whole-genome sequencing, with particular attention to the occurrence of MDR strains. The strains were tested against antibiotics from nine antimicrobial classes (penicillins, cephalosporins, aminoglycosides, tetracyclines, macrolides, fluoroquinolones, lincosamides, phenicols, and sulfonamides). Antimicrobial resistance gene (ARG) sequences and single-nucleotide polymorphisms (SNPs) were evaluated in paired reads using the Bacterial and Viral Bioinformatics Resource Center (BV-BRC) and the Comprehensive Antibiotic Resistance Database (CARD) via Resistance Gene Identifier (RGI), respectively. Results: Phenotypic results indicated that cephalosporins and phenicols were the most effective drugs against P. multocida; however, the majority of strains were also susceptible to fluoroquinolones and tetracyclines. In contrast, high resistance rates were observed to sulfamethoxazole and clindamycin. The most prevalent resistance genes were strA, sul2, and tetH, while none of the strains harbored the bla-_TEM or erm (42) genes. Conclusions: Of the two phenotypic methods, MIC values showed a stronger positive correlation with genotypic results, making it a more suitable method for determining antibiotic susceptibility. The phenotypic results for phenicols, tetracyclines, and fluoroquinolones showed a strong correlation with the detected resistance genes. In contrast, resistance to sulfamethoxazole, β-lactams, and macrolides remained genetically unexplained, suggesting the existence of additional resistance mechanisms to be explored.

Excessive use of antibiotics to control colibacillosis on poultry farms in Morocco contributes to the development of multiresistant avian pathogenic E. coli. Autogenous vaccines are an alternative to antibiotic usage; the main goal of this study was to assess the efficacy of this approach to control recurrent outbreaks of colibacillosis in a large broiler production flow. A tetravalent autogenous vaccine (serotypes O78, O88, O91, and O185) was produced from E. coli strains isolated from clinical lesions in breeding hens and used on two farms (65,000 breeding hens in total). On each farm, contemporaneous groups were included: groups vaccinated twice (12 and 18 weeks of age) and unvaccinated control groups. The performances of these flocks, as well as the performances of broilers hatched from these breeders (12 farms, over 600,000 broilers in total), were recorded. A significant reduction in lesion scores, mortality rate and a significant increase in the number of saleable day-old chicks were observed among vaccinated hens. In broilers, body weight at slaughter, lesion scores, food conversion ratio, mortality and condemnation rates were significantly improved. This is the first field observation of a lasting effect of breeder hens immunization with an autogenous vaccine against E. coli on their offspring.

The classification systems currently in use for Streptococcus suis are primarily based on capsular polysaccharide (CPS) serotyping or multi-locus sequence typing (MLST). A genetic typing system based on the conserved Mac-1 domain of the Ide_Ssuis gene, which encodes an IgM-cleaving protease, could complement these approaches. Specifically, in this study, a genotyping system based on Mac-1 sequence homology was established, qualitative SYBR Green real-time PCR (qPCR) assays for standardized genotype classification were developed, and the global and European distribution of genotypes were analyzed. The Mac-1 classification system identified “two genotypes”, one of which has “two subgenotypes”, based on 427 unique sequences obtained from 4161 global sequences. GT1.1 was the most prevalent, having an amino acid similarity of 97.1%, followed by GT1.2 98.1%, and GT2 91.4%. The qPCR approach enabled reproducible, cross-laboratory-accessible typing. Among 5479 Streptococcus suis European isolates, 64% were GT1.1, 4.5% were GT1.2, and 29.6% were GT2, with a trend toward increasing GT1.1 detection rates between 2018 and 2023. Genotyping classification could complement current Streptococcus suis classification approaches to better support vaccine design and disease control strategies.