Naturally acquired infection of humans with a marine mammal-associated Brucella sp. has only been reported once previously in a study describing infections of two patients from Peru. We report the isolation and characterization of a strain of Brucella from a New Zealand patient that appears most closely related to strains previously identified from marine mammals. The isolate was preliminarily identified as Brucella suis using conventional bacteriological tests in our laboratory. However, the results profile was not an exact match, and the isolate was forwarded to four international reference laboratories for further identification. The reference laboratories identified the isolate as either B. suis or B. melitensis by traditional bacteriological methods in three laboratories and by a molecular test in the fourth laboratory. Molecular characterization by PCR, PCR-restriction fragment length polymorphism, and DNA sequencing of the bp26 gene; IS711; the omp genes omp25, omp31, omp2a, and omp2b; IRS-PCR fragments I, III, and IV; and five housekeeping gene fragments was conducted to resolve the discrepant identification of the isolate. The isolate was identified to be closely related to a Brucella sp. originating from a United States bottlenose dolphin (Tursiops truncatus) and common seals (Phoca vitulina).Brucellosis is an important zoonotic disease of humans, causing a variety of vague symptoms including undulant fever, fatigue, malaise, joint pain, myalgia, depression, and anorexia (22). Chronic sequelae and recrudescence decades after initial infection also occur. Brucella may be transmitted from animals to humans by direct contact with infected animals, ingestion of infected food products, and inhalation of aerosols. Four species of Brucella are the primary causes of infection in humans. Brucella melitensis is highly infectious and is transmitted from sheep and goats, B. abortus is transmitted from cattle, B. suis is transmitted from pigs and, infrequently, B. canis is transmitted from dogs. Other species of Brucella have been rarely or not reported to infect humans.There are only two reports in the literature of humans infected with marine mammal strains of Brucella. One report was of a laboratory worker who displayed symptoms consistent with brucellosis (4). The infection was confirmed by a positive serological response, isolation, and PCR-restriction fragment length polymorphism (RFLP) identification of a marine Brucella strain. Two patients originating from Peru and diagnosed with neurobrucellosis were also confirmed to be infected with marine mammal strains of Brucella by isolation, PCR, and DNA sequencing (41). The two Peruvian patients were not laboratory workers, and the infection was naturally acquired.Serological evidence and isolation of brucellae have been reported from a variety of marine mammals on numerous occasions from locations in the northern hemisphere. The serological prevalence ranges from 0 to 38% for cetaceans, pinnipeds, and mustelids (6,24,26,29,32,34,43). The largest studies of 1,855 ...

Illumina sequencing allows rapid, cheap and accurate whole genome bacterial analyses, but short reads (<300 bp) do not usually enable complete genome assembly. Long-read sequencing greatly assists with resolving complex bacterial genomes, particularly when combined with short-read Illumina data (hybrid assembly). However, it is not clear how different long-read sequencing methods affect hybrid assembly accuracy. Relative automation of the assembly process is also crucial to facilitating high-throughput complete bacterial genome reconstruction, avoiding multiple bespoke filtering and data manipulation steps. In this study, we compared hybrid assemblies for 20 bacterial isolates, including two reference strains, using Illumina sequencing and long reads from either Oxford Nanopore Technologies (ONT) or SMRT Pacific Biosciences (PacBio) sequencing platforms. We chose isolates from the family Enterobacteriaceae, as these frequently have highly plastic, repetitive genetic structures, and complete genome reconstruction for these species is relevant for a precise understanding of the epidemiology of antimicrobial resistance. We de novo assembled genomes using the hybrid assembler Unicycler and compared different read processing strategies, as well as comparing to long-read-only assembly with Flye followed by short-read polishing with Pilon. Hybrid assembly with either PacBio or ONT reads facilitated high-quality genome reconstruction, and was superior to the long-read assembly and polishing approach evaluated with respect to accuracy and completeness. Combining ONT and Illumina reads fully resolved most genomes without additional manual steps, and at a lower consumables cost per isolate in our setting. Automated hybrid assembly is a powerful tool for complete and accurate bacterial genome assembly.

on behalf of the REHAB 7 consortium 8 9

Our results add further evidence for the mobilization of the mcr-pap2 unit from Moraxella via composite transposons leading to its global dissemination. The presence of mcr-pap2 from recent Moraxella isolates indicates they may comprise a reservoir for mcr.

No abstract