Acinetobacter junii: an emerging One Health pathogen.

Acinetobacter junii is an opportunistic human and animal pathogen severely understudied. Here, we conducted the largest genomic epidemiological study on this pathogen to date. Our data show that this bacterium has spread globally. Also, we found that some human and non-human isolates are not well differentiated from one another, implying transmission between clinical and non-clinical, non-human settings. Remarkably, human but also some non-human isolates have clinically important antibiotic resistance genes, and some of these genes are located in plasmids. Given these results, we put forward that A. junii should be considered an emerging One Health problem. In this regard, future molecular epidemiological studies about this species will go beyond human isolates and will consider animal-, plant-, and water-associated environments. IMPORTANCE: Acinetobacter baumannii is the most well-known species from the genus Acinetobacter. However, other much less studied Acinetobacter species could be important opportunistic pathogens of animals, plants and humans. Here, we conducted the largest genomic epidemiological study of A. junii, which has been described as a source not only of human but also of animal infections. Our analyses show that this bacterium has spread globally and that, in some instances, human and non-human isolates are not well differentiated. Remarkably, some non-human isolates have important antibiotic resistance genes against important antibiotics used in human medicine. Based on our results, we propose that this pathogen must be considered an issue not only for humans but also for veterinary medicine.

Identification and characterization of Dutch Avibacterium paragallinarum isolates and the implications for diagnostics.

This study reports the results of diagnostic and molecular typing methods for 18 Avibacterium paragallinarum isolates obtained from outbreaks of infectious coryza in commercial layer flocks in the Netherlands. Isolation, biochemical identification, species-specific PCR tests and classical serotyping were performed. In addition, molecular typing by Enterobacterial Repetitive Intergenic Consensus-Based Polymerase Chain Reaction (ERIC-PCR) and sequence analysis of the partial HPG2 region of A. paragallinarum were applied and results of both techniques were compared. Moreover, the pathogenicity of an isolate of the most common genotype detected in the Netherlands was determined in an animal experiment. All 18 Avibacterium isolates were nicotinamide adenine dinucleotide-dependent. All isolates were detected by the species-specific conventional PCR while 33% of the isolates were missed by the species-specific real-time PCR. Sequence analysis showed a probe mismatch as a result of a single nucleotide polymorphism (G1516A). Modification of the probe of the real-time PCR was necessary to overcome false negative results. Molecular typing showed that sequence analysis of the partial HPG2 region was in concordance with ERIC-PCR results and indicated the presence of two major genotypes. Serotyping showed the presence of serovars A-1, A-2 and B-1. There was no correlation between genotyping results and serotyping results. Inoculation of an isolate of the most prevalent genotype, and belonging to serovar A-1, into brown layer hens demonstrated the pathogenicity of this isolate.

Genomic Islands in the Full-Genome Sequence of an NAD-Hemin-Independent Avibacterium paragallinarum Strain Isolated from Peru.

Here, we report the full-genome sequence of an NAD-hemin-independent Avibacterium paragallinarum serovar C-2 strain, FARPER-174, isolated from layer hens in Peru. This genome contained 12 potential genomic islands that include ribosomal protein-coding genes, a nadR gene, hemocin-coding genes, sequences of fagos, an rtx operon, and drug resistance genes.

Phylogenetic relationship of Ornithobacterium rhinotracheale strains.

The bacterium Ornithobacterium rhinotracheale is associated with respiratory disease in wild birds and poultry. In this study, the phylogenetic analysis of nine reference strains of O. rhinotracheale belonging to serovars A to I, and eight Mexican isolates belonging to serovar A, was performed. The analysis was extended to include sequences from another 23 strains available in the public domain. The analysis showed that the 40 sequences formed six clusters, I to VI. All eight Mexican field isolates were placed in cluster I. One of the reference strains appears to present genetic diversity not previously recognized and was placed in a new genetic cluster. In conclusion, the phylogenetic analysis of O. rhinotracheale strains, based on the 16S rRNA gene, is a suitable tool for epidemiologic studies.

Isolation and molecular characterization of Corynebacterium pseudotuberculosis from sheep and goats in Mexico.

The pathogenic bacteria of Corynebacterium pseudotuberculosis caused a chronic contagious infectious disease of the caseous lymphadenitis or pseudotuberculosis. Globally, isolates obtained from different injuries that affect sheep and goats, have been identified by fully or partially gene sequencing. However, in Mexico there is no complete study to identify by molecular and phylogenetic techniques the circulating isolates as well as its virulence factors. Therefore, in the present study we reported the identification of 57 isolates of C. pseudotuberculosis by bacteriological tests and the amplification of 16S rRNA, rpoB and pld genes, as well as, genes involved in virulence and pathogenicity: Fag A, Fag B, Fag C, Fag D and hsp60. Phylogenetic analysis was performed based on the partial sequence of the rpoB gene. Genes involved in virulence and pathogenicity were identified in the 98.2% of the isolates. Regarding the phylogenetic analysis, were identified the species and subspecies to which they belong of all the tested isolates. The phenotypic and genotypic characterization will allow to establish preventive and prophylactic measures aimed to the creation of effective immunogens against Corynebacterium pseudotuberculosis.

Serotyping, Genotyping, and Antimicrobial Susceptibility of Ornithobacterium rhinotracheale Isolates from Mexico.

The bacterium Ornithobacterium rhinotracheale is associated with respiratory disease and septicemia in poultry. In this study, 9 reference strains and a total of 23 isolates of O. rhinotracheale from respiratory diseased poultry from Mexico were serotyped and genotyped. Furthermore, the antimicrobial susceptibility of isolates and reference strains of O. rhinotracheale were determined. All isolates belong to serotype A and showed a clonal relationship. All reference strains and isolates were resistant to colistin, fosfomycin, gentamicin, kanamycin, streptomycin, and trimethoprim-sulfamethoxazole. These results should eventually be helpful in planning strategies for the control of O. rhinotracheale infections in poultry in Mexico.

An evaluation of serotyping of Avibacterium paragallinarum by use of a multiplex polymerase chain reaction.

In the present study, the ability of a recently proposed multiplex polymerase chain reaction (mPCR) to determine the serogroups (A, B, and C) of Avibacterium paragallinarum was evaluated. A total of 12 reference strains and 69 field isolates of Av. paragallinarum from Ecuador, Mexico, Panama, and Peru were included in the study. With some exceptions (which were serotyped in the current study), all of the isolates and strains had been previously examined by 2 serotyping schemes (Page and Kume) or were the formal reference strains for the schemes. Three of 6 (50%) reference strains of serogroup A, 2 (100%) of serogroup B, and 1 of 4 (25%) reference strains of serogroup C were correctly serotyped by the mPCR. With the field isolates, the mPCR correctly recognized 16 of the 17 serogroup A isolates, 10 of the 12 serogroup B isolates, and 18 of the 37 serogroup C isolates. Overall, the specificity and sensitivity of the PCR test was as follows: 82.6% and 87.3% (serogroup A), 85.7% and 71.9% (serogroup B), and 46.3% and 100% (serogroup C). The poor performance of the mPCR in terms of recognition of serogroup C isolates (low sensitivity of 46.3%) and the relatively high level of uncertainty about the accuracy of the serogroup A and B results (specificity of 87.3% and 71.9%, respectively) means that the assay cannot be recommended as a replacement for conventional serotyping.

Aislamiento e identificación de Avibacterium paragallinarum NAD independiente (serovariedad C-1) en gallinas de reemplazo con coriza infecciosa / Isolation and identification of NAD-independent Avibacterium paragallinarum (serovar C-1) from growing layer hens with infectious coryza

The isolation of V-factor (NAD)-independent, serovar C-1 of Avibacterium paragallinarum from growing layer hens with infectious coryza, is reported for the first time. Av. paragallinarum serovar C-1 isolates were PCR confirmed and shared a unique genetic profile. Isolation of Av. paragallinarum NAD-independent biovar, serotyping and genotyping of isolates at the avian diagnostic laboratories are discussed.

Se informa por primera ocasión el aislamiento de Avibacterium paragallinarum, serovariedad C-1, independiente del factor V de crecimiento (NAD), a partir de gallinas de reemplazo con coriza infecciosa. Los aislamientos de Av. paragallinarum serovariedad C-1 incluidos en el estudio fueron confirmados por PCR y compartieron un perfil genético único. Se discute considerar en los laboratorios de diagnóstico aviar el aislamiento de Av. paragallinarum de la biovariedad independiente de NAD, así como la necesidad de serotipificación y tipificación genética de los aislamientos.

Hemagglutinin serotyping of Avibacterium paragallinarum isolates from Ecuador.

Avibacterium paragallinarum is the causative agent of infectious coryza, an acute respiratory disease of chickens. In this study, a total of 28 isolates of A. paragallinarum from Ecuador were serotyped by the hemagglutinin scheme which recognizes nine serovars. Out of 28 isolates, 17 isolates belonged to serovar A-3, and five isolates to each serovars B-1 and C-1, whereas one isolate was non-typeable. This is the first report of A. paragallinarum serovar A-3 outside Brazil and serovar C-1 outside Japan.