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The pharmaceutical industry must comply with the requirements for good manufacturing practices to reduce inherent contamination risks in the production process. Bacillus and related genera are among the main bacterial isolated from clean areas, raw material, and products in pharmaceutical industries, but the correct identification of these species is still a challenge. The aim of this study was to characterize by phenotyping, protein profiling, and 16S rRNA gene sequencing S. horikoshii strains (n = 6) isolated from an immunobiological pharmaceutical facility, and to propose the reclassification of Bacillus tianshenii to the genus Sutcliffiella, and Sutcliffiella tianshenii sp. nov. The strains were characterized by VITEK®2, Matrix-Assisted Laser Desorption Ionization-Time of Flight/Mass Spectrometry (MALDI-TOF/MS) using VITEK®MS, and 16S rRNA gene sequencing analysis. MALDI-TOF/MS did not identify any strains, that were identified by 16S rRNA as S. horikoshii. VITEK®2 showed false-positive results, with misidentification as Bacillus sporothermodurans (reclassified as Heyndrickxia sporothermodurans) and Geobacillus thermoleovorans. After MALDI-TOF/MS database expansion, with the creation of SuperSpectrum, the strains were correctly identified as S. horikoshii. This study is the first report of isolation of S. horikoshii strains from a pharmaceutical industry. More studies are necessary to better understand the ability of S. horikoshii to contaminate the environment and products.

Introduction: Bioburden monitoring is essential to contamination control of pharmaceutical products. Microbiological load may represent a potential risk for patients if the sterilization process is not effective and/or due to the presence of allergens or toxins. Although bioburden can be eliminated by terminal sterilization or filtration processes, it is important to monitor it before final processing, regarding the amount and species of microorganisms that are present. Objective:To evaluate the microorganism's species isolated in intermediate process solutions samples to be used in immunobiologicals formulations from a pharmaceutical industry before sterilizing by filtration.Methodology: All the microorganisms isolated from the bioburden test in the period from January 2018 to December 2020 were evaluated. They were classified into three groups according to the process criticality: Group 1-critical microorganisms listed in the Brazilian and International Pharmacopoeias (Escherichia coli, Pseudomonas aeruginosa, Salmonella, Staphylococcus aureus and Candida albicans); Group 2-questionable microorganisms that can be a risk to the process; Group 3-non-questionable microorganisms, that are not a risk to the process. The Group 2 microorganisms were evaluated according to the following criteria: indication of fecal contamination, pathogenicity for humans, and production of toxins that can be perpetuated throughout the process with risk to patients. Microorganisms identified as human pathogens have been classified into three categories: 1)pathogens that can cause infections in immunocompetent individuals; 2)opportunists that can show virulence and are responsible for infections in immunocompromised or debilitated individuals; 3)exceptional opportunists that rarely cause infections in humans.Results: 78 generas and 171 species were isolated: two species (1.2%) were classified in Group 1, 135 (78.9%) in Group 2, and 34 (19.9%) in Group 3. The two Group 1 isolated species were S.aureus and P.aeruginosa. Among the 135 Group 2 species, 92.6% (n=125) were bacteria and 7.4% (n=10) were yeasts and molds. Thirteen species (9.6%) were fecal contamination indicators. One hundred and thirty-four (99.3%) species were identified as human pathogens, but 4.5% (n=6) belonged to category 1; 38.8% (n=52) to category 2 and 56.7% (n=76) to category 3. Twenty-eight species (20.7%) were known for producing various toxins; and 40.7% (n=55) were endotoxins producers. In Group 3, there were 97.1% (n=33) bacteria, mainly identified in Order Caryophanales, and 2.9% (n=1) fungi identified as Lecanicillium coprophilum. In the case of Group 1 bacteria's, the intermediate process solution was immediately discarded and an investigation was initiated to identify the root cause and subsequently the adoption of corrective and preventive actions. In the case of Group 2 microorganisms, a risk assessment for each specific process was carried out in order to decide whether this risk could be mitigated or eliminated along the production chain, so as not to po...

Os fungos são microrganismos que podem representar risco durante a produção de medicamentos nas indústrias farmacêuticas. A correta identificação desses contaminantes é de grande importância para auxiliar na análise de riscos e na tomada de ações preventivas e corretivas durante a cadeia produtiva. O objetivo deste estudo foi realizar uma revisão integrativa da literatura referente à ocorrência de fungos filamentosos em indústrias farmacêuticas e das metodologias utilizadas para sua identificação. Os gêneros mais ocorrentes identificados foram: Aspergillus, Cladosporium, Penicillium e Fusarium, sendo relatados em trabalhos de três países. As metodologias de identificação encontradas foram: métodos morfológicos (avaliação dos aspectos macroscópicos e microscópicos), caracterização fenotípica, análise proteômica por MALDI-TOF MS e análise de sequencias de DNA das regiões ITS e D1/D2 do DNA ribossomal. Poucos estudos relativos à identificação de fungos filamentosos e a sua diversidade em indústrias farmacêuticas foram identificados na literatura consultada, indicando que este tema precisa ser mais explorado.