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We investigated Bordetella holmesii and Bordetella pertussis in 935 suspected pertussis cases in northern India (2019-2023) using PCR and serology. B. holmesii showed increased prevalence in pertussis cases, particularly in older children, highlighting its emerging role and the need for ongoing surveillance and adjusted prevention strategies.

To determine changes in Bordetella pertussis and B. parapertussis detection rates, we analyzed 1.43 million respiratory multiplex PCR test results from US facilities from 2019 through mid-2023. From mid-2022 through mid-2023, Bordetella spp. detection increased 8.5-fold; 95% of detections were B. parapertussis. While B. parapertussis rates increased, B. pertussis rates decreased.

Little is known about oxygen utilization during infection by bacterial respiratory pathogens. The classical Bordetella species, including B . pertussis , the causal agent of human whooping cough, and B . bronchiseptica , which infects nearly all mammals, are obligate aerobes that use only oxygen as the terminal electron acceptor for electron transport-coupled oxidative phosphorylation. B . bronchiseptica , which occupies many niches, has eight distinct cytochrome oxidase-encoding loci, while B . pertussis , which evolved from a B . bronchiseptica -like ancestor but now survives exclusively in and between human respiratory tracts, has only three functional cytochrome oxidase-encoding loci: cydAB1 , ctaCDFGE1 , and cyoABCD1 . To test the hypothesis that the three cytochrome oxidases encoded within the B . pertussis genome represent the minimum number and class of cytochrome oxidase required for respiratory infection, we compared B . bronchiseptica strains lacking one or more of the eight possible cytochrome oxidases in vitro and in vivo . No individual cytochrome oxidase was required for growth in ambient air, and all three of the cytochrome oxidases conserved in B . pertussis were sufficient for growth in ambient air and low oxygen. Using a high-dose, large-volume persistence model and a low-dose, small-volume establishment of infection model, we found that B . bronchiseptica producing only the three B . pertussis -conserved cytochrome oxidases was indistinguishable from the wild-type strain for infection. We also determined that CyoABCD1 is sufficient to cause the same level of bacterial burden in mice as the wild-type strain and is thus the primary cytochrome oxidase required for murine infection, and that CydAB1 and CtaCDFGE1 fulfill auxiliary roles or are important for aspects of infection we have not assessed, such as transmission. Our results shed light on the environment at the surface of the ciliated epithelium, respiration requirements for bacteria that colonize the respiratory tract, and the evolution of virulence in bacterial pathogens.

Bordetella pertussis causes the acute disease whooping cough and survives only in the human respiratory tract, while Bordetella bronchiseptica causes long-term, chronic infections in a broad range of mammals and can also survive in extra-host environments. These bacteria produce a nearly identical set of virulence factors, including adenylate cyclase toxin (ACT), a protein that is modified by the addition of acyl chains. Acylation is required for ACT to cause hemolysis and for efficient intoxication of host cells in vitro . We found that ACT acylation is also important, but not absolutely required, during infection. We also discovered differences in ACT production, acylation, and secretion between B. bronchiseptica and B. pertussis that may contribute to the different virulence strategies of these species. This study highlights the advantage of conducting comparative analyses between closely related species to better understand the evolution of virulence.

Background Niviventer , a rodent species widely distributed in Asian forests, serves as a significant reservoir for pathogens. Bordetella pseudohinzii ( B. pseudohinzii ), a recently identified Bordetella species with unclear pathogenic potential, poses challenges in species identification and understanding of its pathogenicity, its biological traits and antibiotic resistance are not well understood. Methods B. pseudohinzii (strains 21F10, 22F12, and 27F25) were isolated from lung tissue of wild niviventer rodents in Guizhou, China. Initial identification was performed using Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) and 16 S rRNA gene sequencing. A phylogenetic tree based on the 16 S rRNA gene sequences was constructed using the neighbor-joining method implemented in MEGA 11. Whole-genome sequencing (WGS) was conducted on all three strains, and strain 21F10 underwent hybrid assembly of second- and third-generation sequencing to achieve high-quality sequences. Average Nucleotide Identity (ANI) and digital DNA-DNA hybridization (dDDH) were used as gold standards for strain identification, with thresholds set at 95% and 70%, respectively. Gene annotation was performed using nine databases, including KEGG, VFDB, CARD, PHI, COG, and NR. Antimicrobial susceptibility testing was carried out using the drug-sensitive plate method. Results Initial MALDI-TOF MS identification misclassified the strains as B. avium and B. hinzii . However, PCR amplification of the 16 S rRNA gene (primers 27 F and 1492R) revealed that the strains were identified as B. hinzii (identity > 99%). Further analysis of the 16 S rRNA gene sequences obtained from WGS showed identities greater than 99% with both B. pseudohinzii and B. hinzii . Phylogenetic analysis of the 16 S rRNA gene sequences showed that the strains were closely related to B. hinzii , followed by B. pseudohinzii . Ultimately, the ANI values of all three strains with B. pseudohinzii were greater than 95%, and dDDH values exceeded 70%, confirming the strains as B. pseudohinzii . Strain 21F10 exhibited notable findings in terms of virulence factors and antibiotic resistance genes. Antimicrobial susceptibility testing revealed significant resistance to several cephalosporins (cefoxitin, cefuroxime, cefotaxime, cefazolin, and ceftiofur). The 16 S rRNA and WGS of strain 21F10 have been deposited in GenBank and Genome Sequence Archive (GSA)under accession numbers PQ881859 and CRA022358, respectively. Conclusion The first isolation of B. pseudohinzii from the lung tissue of wild niviventer was reported, and the limitations of traditional methods for identifying B. pseudohinzii were demonstrated. We highlight the superiority of WGS for accurate species identification. The findings reveal a complex pathogenic profile and notable antibiotic resistance, providing important insights for the future prevention and treatment of B. pseudohinzii infections in humans, as well as underscoring the need for monitoring B. pseudohinzii in rodent populations. Clinical trial number Not applicable.

Whooping cough is resurging in the United States despite high vaccine coverage. The rapid rise of Bordetella pertussis isolates lacking pertactin (PRN), a key vaccine antigen, has led to concerns about vaccine-driven evolution. Previous studies showed that pertactin can mediate binding to mammalian cells in vitro and act as an immunomodulatory factor in resisting neutrophil-mediated clearance. To further investigate the role of PRN in vivo , we examined the functions of pertactin in the context of a more naturally low dose inoculation experimental system using C3H/HeJ mice that is more sensitive to effects on colonization, growth and spread within the respiratory tract, as well as an experimental approach to measure shedding and transmission between hosts. A B . bronchiseptica pertactin deletion mutant was found to behave similarly to its wild-type (WT) parental strain in colonization of the nasal cavity, trachea, and lungs of mice. However, the pertactin-deficient strain was shed from the nares of mice in much lower numbers, resulting in a significantly lower rate of transmission between hosts. Histological examination of respiratory epithelia revealed that pertactin-deficient bacteria induced substantially less inflammation and mucus accumulation than the WT strain and in vitro assays verified the effect of PRN on the induction of TNF-α by murine macrophages. Interestingly, only WT B . bronchiseptica could be recovered from the spleen of infected mice and were further observed to be intracellular among isolated splenocytes, indicating that pertactin contributes to systemic dissemination involving intracellular survival. These results suggest that pertactin can mediate interactions with immune cells and augments inflammation that contributes to bacterial shedding and transmission between hosts. Understanding the relative contributions of various factors to inflammation, mucus production, shedding and transmission will guide novel strategies to interfere with the reemergence of pertussis.

Introduction Cunninghamella elegans infections cause rare and severe mucormycosis. Bordetella bronchiseptica and Pneumocystis jirovecii relate to pneumonia. They are all clinically uncommon pathogens and no reports of co-infections have been reported. Case presentation Here we present a case of a 67-year-old male patient who initially presented with fever, chills, and mild cough. B. bronchiseptica , P. jirovecii , Aspergillus fumigatus , and human alphaherpesvirus 1 (HSV1) were detected by clinical metagenomic next-generation sequencing (mNGS) of his bronchoalveolar lavage fluid (BLAF). Despite receiving anti-infective treatment, the patient rapidly developed respiratory failure and was transferred to the intensive care unit. Subsequent mNGS testing further revealed the presence of C. elegans , indicating that different pathogens played dominant roles at various stages of the disease progression. The routine culture also identified several of the above pathogens, but the results were reported much later than those of mNGS. Eventually, imaging findings and symptoms of the patient improved with comprehensive antibiotic coverage, and he was transferred to a lower-level hospital for rehabilitation treatment. Conclusions This is the first detailed report of the combined infection of B. bronchiseptica , P. jirovecii , and C. elegans . During the treatment process, we also observed rare and unusual neurological side effects: visual and auditory hallucinations, restlessness, and aphasia. Also, the case indicates that traditional methods are insufficient for the etiological diagnosis needs of critical and severe patient populations, and timely use of mNGS should be recommended.

An increase in positive Bordetella parapertussis tests among patients in a teaching hospital in the Netherlands resulted in enhanced infection control and microbiological surveillance. Further analysis revealed that batches of contaminated nasopharyngeal swabs were associated with a pseudo-outbreak, resulting in incorrect diagnoses, antimicrobial treatments, isolation precautions, and public health notifications.

Bordetella bronchiseptica is a potential zoonotic pathogen, which mainly causes respiratory diseases in humans and a variety of animal species. B. bronchiseptica is one of the important pathogens isolated from rabbits in Fujian Province. However, the knowledge of the epidemiology and characteristics of the B. bronchiseptica in rabbits in Fujian Province is largely unknown. In this study, 219 B. bronchiseptica isolates recovered from lung samples of dead rabbits with respiratory diseases in Fujian Province were characterised by multi-locus sequencing typing, screening virulence genes and testing antimicrobial susceptibility. The results showed that the 219 isolates were typed into 11 sequence types (STs) including five known STs (ST6, ST10, ST12, ST14 and ST33) and six new STs (ST88, ST89, ST90, ST91, ST92 and ST93) and the ST33 (30.14%, 66/219), ST14 (26.94%, 59/219) and ST12 (16.44%, 36/219) were the three most prevalent STs. Surprisingly, all the 219 isolates carried the five virulence genes (fhaB, prn, cyaA, dnt and bteA) in the polymerase chain reaction screening. Moreover, the isolates were resistant to cefixime, ceftizoxime, cefatriaxone and ampicillin at rates of 33.33%, 31.05%, 11.87% and 3.20%, respectively. This study showed the genetic diversity of B. bronchiseptica in rabbits in Fujian Province, and the colonisation of the human-associated ST12 strain in rabbits in Fujian Province. The results might be useful for monitoring the epidemic strains, developing preventive methods and preventing the transmission of epidemic strains from rabbits to humans.

Bordetella pertussis is the etiologic agent of whooping cough and has been the target of vaccination for over fifty years. The latest strategies include the use of acellular pertussis vaccines that induce specific immunity against few virulence factors amongst which pertactin is included in three and five component acellular pertussis vaccines. Recently, it has been reported that B. pertussis clinical isolates loose the production of this adhesin in regions reaching high vaccine coverage with vaccines targeting this virulence factor. We here demonstrate that isolates not producing pertactin are capable of sustaining longer infection as compared to pertactin producing isolates in an in vivo model of acellular pertussis immunization. Loosing pertactin production might thus provide a selective advantage to these isolates in this background, which could account for the upraise in prevalence of these pertactin deficient isolates in the population.