Characterization of Pasteurella multocida isolated from ducks in China from 2017 to 2019

Pasteurella multocida is a leading cause of respiratory disorders in pigs. This study was designed to understand the genotypical and antimicrobial resistant characteristics of P. multocida from pigs in China. To achieve this, we briefly investigated 158 P. multocida isolates from pigs with respiratory disorders in China between 2019 and 2020. Genotyping through multiplex PCR assays assigned these 158 isolates into capsular genotypes A (60.13%, 95/158), D (35.44%, 56/158), F (4.43%, 7/158), and/or lipopolysaccharide (LPS) genotypes L3 (28.48%, 45/158) and L6 (66.46%, 105/158). In addition, eight isolates (5.06%, 8/158) were found to be nontypable using the LPS genotyping method. When combining the capsular genotypes and the LPS genotypes, D: L6 (34.81%, 55/158) and A: L6 (31.65%, 50/158) were the predominant genotypes, followed by A: L3 (24.05%, 38/158). PCR detection of virulence factor-encoding genes showed that over 80% of the isolates were positive for exbB, tonB, exbD, ompH, ptfA, fimA, sodA, sodC, fur, ompA, oma87, plpB, hsf-2, nanH and hgbB, suggesting the presence of these genes were broad characteristics of P. multocida. We also found approximately 63.92% (101/158), 51.27% (81/158), 8.86% (14/158), 7.59% (12/158), 3.16% (5/158), 0.63% (1/158), and 0.63% (1/158) of the isolates grew well in media with the presence of colistin (4 μg/mL), tetracycline (16 μg/mL), tigecycline (1 μg/mL), ampicillin (32 μg/mL), chloramphenicol (32 μg/mL), cefepime (16 μg/mL), and ciprofloxacin (1 μg/mL), respectively. This study contributes to the understanding of genotypes and antimicrobial resistance profile of P. multocida currently circulation in pigs of China. Supplementary information The online version contains supplementary material available at 10.1186/s44149-021-00031-7.

Pasteurella multocida capsular: lipopolysaccharide types D:L6 and A:L3 remain to be the main epidemic genotypes of pigs in China.

The number of pets such as dogs, cats, rabbits, and parrots has increased in European families. Social benefits to owners such as decreasing feelings of loneliness and anxiety are provided by pets which are also used in Animal-assisted Therapy (AAT). Nevertheless, human-animal interactions are also associated with health problems including allergies, asthma, and zoonosis. Rabbits may carry potential pathogens for humans. One of the most common bacteria that colonizes the oro-pharynx and the upper respiratory tract of rabbits is Pasteurella (P.) multocida. Transmission of the infection to humans results from scratches, licks, and bites but it also can occur from the inhalation of air particles containing the microorganism. Immunocompromised people or persons with pulmonary disorders are particularly susceptible to the infection. Infected rabbits may carry P. multocida with or without clinical signs. In this paper, the sensitivity to antibiotics and the invasiveness ability of P. multocida identified in a farm of pet rabbits affected by severe pasteurellosis were investigated. The strain was P. multocida belonging to capsular type A which is the type most often detected in humans. The identified strain was susceptible to the tested antibiotics, but it appeared equipped with several virulence genes which are responsible for fimbriae production, adhesion processes to host cells, enzyme production, and are involved in iron acquisition processes. These findings are of particular interest because rabbits recovered from pasteurellosis very often become carriers of the bacteria. Therefore, we suggest considering P. multocida screening in the routine medical checks of rabbits, especially if they are meant to be companion animals for children and elder people, given that the transmission of the pathogen cannot be excluded.

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Spreading of Pasteurella multocida Infection in a Pet Rabbit Breeding and Possible Implications on Healed Bunnies

Naked-eye on-site detection platform for Pasteurella multocida based on the CRISPR-Cas12a system coupled with recombinase polymerase amplification.

Simple Summary Bacterial and viral respiratory diseases are a major health problem in pig farms. Most often multiple infectious agents are involved in the development of clinical cases of respiratory diseases. It is necessary to conduct laboratory etiological tests on suspected cases. In January 2021, a 70-day-old pig herd in a pig farm in Hebei Province, China, had been suffering from respiratory problems. On the basis of this preliminary clinical diagnosis, this study carried out comprehensive laboratory detection including nucleic acid detection and sequencing, the isolation and identification of the pathogen, genetic evolution analysis, and genotype or serotype identification, as well as an analysis of the bacterial morphology characteristics and pathogenicity. The results showed that the co-infection of PCV2a and P. multocida capsular serotypes D was likely responsible for the disease. This study could provide important etiological information for effective control and treatment of the disease in pig farms. Abstract In order to diagnose a respiratory disease in a pig farm, the lungs, spleen, and lymph nodes of three dead pigs were collected for pathogen detection by PCR and isolation on the basis of preliminary clinical diagnosis. The virus isolate was identified by gene sequence analysis and Immunoperoxidase monolayer assay (IPMA). The bacterial isolate was identified by biochemical tests, 16S rDNA sequence analysis, and species- and serotype-specific PCR, and the pathogenicity was analyzed. Porcine circovirus type 2a (PCV2a) genotype from the lungs, spleen, and lymph nodes and Pasteurella (P.) multocida capsular serotypes D from the lungs were found. The PCV2a isolates could specifically bound the anti-PCV2-Cap polyclonal antibody. The 16S rDNA sequence of P. multocida isolates had 99.9% identity with that of the strain from cattle, and the isolate was highly pathogenic to mice. The results showed that the co-infection of PCV2a and P. Multocida capsular serotypes D should be responsible for the disease. The uncommon PCV2a is still prevalent in some pig farms besides the dominant PCV2d genotype. This study could provide important etiological information for effective control and treatment of the disease in pig farms.

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Detection of Porcine Circovirus Type 2a and Pasteurella multocida Capsular Serotype D in Growing Pigs Suffering from Respiratory Disease

Pasteurella multocida is a bacterial pathogen with the ability to infect a multitude of hosts including humans, companion animals, livestock, and wildlife. This study used bioinformatic approaches to explore the genomic diversity of 656 P. multocida isolates and epidemiological associations between host factors and specific genotypes. Isolates included in this study originated from a variety of hosts, including poultry, cattle, swine, rabbits, rodents, and humans, from five different continents. Multi-locus sequence typing identified 69 different sequence types. In-silico methodology for determining capsular serogroup was developed, validated, and applied to all genome sequences, whereby capsular serogroups A, B, D, and F were found. Whole genome phylogeny was constructed from 237,670 core single nucleotide variants (SNVs) and demonstrated an overall lack of host or capsular serogroup specificity, with the exception of isolates from bovine sources. Specific SNVs within the srlB gene were identified in P. multocida subsp. septica genomes, representing specific mutations that may be useful for differentiating one of the three known subspecies. Significant associations were identified between capsular serogroup and virulence factors, including capsular serogroup A and OmpH1, OmpH3, PlpE, and PfhB1; capsular serogroup B and HgbA and PtfA; and capsular serogroup F and PtfA and PlpP. Various mobile genetic elements were identified including those similar to ICEPmu1, ICEhin1056, and IncQ1 plasmids, all of which harbored multiple antimicrobial resistance-encoding genes. Additional analyses were performed on a subset of 99 isolates obtained from turkeys during fowl cholera outbreaks from a single company which revealed that multiple strains of P. multocida were circulating during the outbreak, instead of a single, highly virulent clone. This study further demonstrates the extensive genomic diversity of P. multocida, provides epidemiological context to the various genotyping schemes that have traditionally been used for differentiating isolates, and introduces additional tools for P. multocida molecular typing.

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Genomic diversity and molecular epidemiology of Pasteurella multocida.

https://doi.org/10.1016/j.psj.2022.102009

Marissa Studniski

Papers

Genomic diversity and molecular epidemiology of Pasteurella multocida.

Refining the definition of the avian pathogenic Escherichia coli (APEC) pathotype through inclusion of high-risk clonal groups