Anovaginal Colonization by Group B Streptococcus and Streptococcus anginosus among Pregnant Women in Brazil and Its Association with Clinical Features.

Streptococcus agalactiae (Group B Streptococcus; GBS) is a leading cause of neonatal invasive disease worldwide. GBS can colonize the human gastrointestinal and genitourinary tracts, and the anovaginal colonization of pregnant women is the main source for neonatal infection. Streptococcus anginosus, in turn, can colonize the human upper respiratory, gastrointestinal, and genitourinary tracts but has rarely been observed causing disease. However, in the last years, S. anginosus has been increasingly associated with human infections, mainly in the bloodstream and gastrointestinal and genitourinary tracts. Although anovaginal screening for GBS is common during pregnancy, data regarding the anovaginal colonization of pregnant women by S. anginosus are still scarce. Here, we show that during the assessment of anovaginal GBS colonization rates among pregnant women living in Rio de Janeiro, Brazil, S. anginosus was also commonly detected, and S. anginosus isolates presented a similar colony morphology and color pattern to GBS in chromogenic media. GBS was detected in 48 (12%) while S. anginosus was detected in 17 (4.3%) of the 399 anovaginal samples analyzed. The use of antibiotics during pregnancy and history of urinary tract infections and sexually transmitted infections were associated with the presence of S. anginosus. In turn, previous preterm birth was associated with the presence of GBS (p < 0.05). The correlation of GBS and S. anginosus with relevant clinical features of pregnant women in Rio de Janeiro, Brazil, highlights the need for the further investigation of these important bacteria in relation to this special population.

Candida palmioleophila: A New Emerging Threat in Brazil?

Human activity directly or indirectly causes climate change, promoting changes in the composition of the atmosphere. This change is beyond the variation of the natural climate. In this manner, climate change could create an environmental pressure which is enough to trigger new fungal diseases. In addition to climate alterations, the onset of the COVID-19 pandemic has also been associated with the emergence of fungal pathogens. Fungi showed that an inability to grow at high temperatures limits the capacity of fungi to infect mammals. However, fungi can develop thermotolerance, gradually adapting to rising temperatures due to climate change, and generating a greater number of disease-causing organisms. In the present study, we reported the detection and identification of Candida palmioleophila isolates recovered from raw sewage samples in Niteroi city, Rio de Janeiro State, Brazil, during a monitoring program for measuring SARS-CoV-2 presence and concentration. Using polyphasic taxonomy to identify the species and evaluating some virulence aspects of this species, such as biofilm formation and extracellular enzyme production, our data highlight this species as a possible emerging pathogen in Brazil, especially in the pandemic context.

Changes in Group B Streptococcus Colonization among Pregnant Women before and after the Onset of the COVID-19 Pandemic in Brazil.

Group B Streptococcus (GBS) is a leading cause of neonatal infections. The genitourinary and gastrointestinal tract of pregnant women are the main source of transmission to newborns. This work investigated the prevalence and characterized GBS from pregnant women in Rio de Janeiro, Brazil, comparing the periods before (January 2019 to March 2020; 521) and during (May 2020 to March 2021; 285) the COVID-19 pandemic. GBS was detected in 10.8% of anovaginal samples. Considering scenarios before and during the pandemic, GBS colonization rate significantly decreased (13.8% vs. 5.3%; p = 0.0001). No clinical and sociodemographic aspect was associated with GBS carriage (p > 0.05). A total of 80%, 13.8% and 4.6% GBS strains were non-susceptible to tetracycline, erythromycin and clindamycin, respectively. Serotype Ia was the most frequent (47.7%), followed by V (23.1%), II (18.4%), III (7.7%) and Ib (3.1%). An increasing trend of serotypes Ib and V, as well as of antimicrobial resistance rates, and a decreasing trend of serotypes II and III, were observed after the pandemic onset, albeit not statistically significant (p > 0.05). The reduction in GBS colonization rates and alterations in GBS serotypes and resistance profiles during the pandemic were not due to changes in the sociodemographic profile of the population. Considering that control and preventive measures related to the COVID-19 pandemic onset have impacted other infectious diseases, these results shed light on the need for the continuous surveillance of GBS among pregnant women in the post-pandemic era.

Influence of Penicillin on Biofilm Formation by Streptococcus agalactiae Serotype Ia/CC23.

Streptococcus agalactiae (Group B Streptococcus , GBS) is a major agent of perinatal infections. Biofilms have been associated with GBS colonization and disease, as well as with infection persistence and recurrence. Although GBS remains susceptible to beta-lactams, it is still unknown how sessile cells respond to these antibiotics. Here, we evaluated the effect of different concentrations of penicillin (3-48 mg/L) on in vitro biofilm formation by four GBS strains belonging to serotype Ia/clonal complexes23 that were recovered from the oropharynx or urine of pregnant women and were previously characterized as strong biofilm producers. All four GBS strains were fully susceptible to penicillin (minimum inhibitory concentration = 0.023 mg/L), but penicillin was not able to fully prevent biofilm formation by these GBS strains. Biofilms formed in the presence of penicillin had reduced biomasses and thickness, but they were still classified as strong. Penicillin significantly reduced the density of live cells, but higher penicillin concentrations did not lead to improved prevention of biofilm formation. Biofilms formed in the presence of penicillin had no channels or long cocci chains observed in penicillin-free biofilms. Overall, results highlight the concerning possible impacts of biofilm formation in penicillin-based treatment and preventive strategies of GBS infections, even when the bacterial strain involved is fully antibiotic-susceptible.

Unusual finding of the human-adapted hypervirulent serotype III/ST17 clone in a historical bovine Group B Streptococcus isolate from Brazil.

Group B Streptococcus (GBS) is a leading cause of human neonatal infections and bovine mastitis. We report here the unusual finding of the human-adapted hypervirulent serotype III/ST17 clone in a bovine GBS isolated in 1987 in Brazil. This isolate shared several phenotypic and genotypic characteristics with serotype III/ST17 strains obtained from human sources, including PFGE pattern, pilus genes, lactose fermentation, DNase activity, and antimicrobial susceptibility profile, highlighting the importance of continued tracking of GBS in the One Health scope. The study brings new evidence for the potential interspecies transmission and sheds new light into evolution aspects of the pathogen Group B Streptococcus (GBS) by reporting the occurrence of an ancient bovine GBS isolate belonging to a variant currently known to be exclusively found in human hosts.

Going Online in Pandemic Time: A DivulgaMicro Workshop Experience.

DivulgaMicro is a Brazilian science communication and outreach project run by three young female scientists. In 2020, due to the COVID-19 pandemic, we promoted the first virtual edition of the DivulgaMicro Workshop. Here, we describe adaptations implemented to deliver the virtual edition and its assessment by the participants. We offered a 3-day workshop addressing manuscript writing and publishing, poster design and oral presentations, and strategies for effectively communicating with the lay public on Days 1, 2, and 3, respectively. Each daily session was divided into short lectures and active learning through interactive and cooperative activities. We applied pre- and postsession surveys daily to assess participants' learning, with an opinion questionnaire at the end of the workshop. We selected 50 of 221 applicants, but only 38 of those selected participated in the workshop. Correct answers were significantly higher (p < 0.01) in the postsession surveys each day. Most of the 34 participants who answered the opinion questionnaire (97%) would recommend the workshop to their colleagues. Participants stated that the workshop content was transmitted in a clear and straightforward manner, and they considered the online format suitable for knowledge acquisition. Participants were satisfied with the organization, dynamics, and interactivity. Topics addressed on Day 3 (61.8%) and Day 1 (23.5%) were most liked and were considered most useful in the participants' careers. We believe that the overall success of the workshop is due to the combination of short lectures with active-learning activities, the use of virtual platforms that enabled effective communication between participants and instructors, and the support of our collaborators.

MALDI-TOF MS: An alternative approach for ribotyping Clostridioides difficile isolates in Brazil.

Clostridioides difficile is an important organism causing healthcare-associated infections. It has been documented that specific strains caused multiple outbreaks globally, and patients infected with those strains are more likely to develop severe C. difficile infection (CDI). With the appearance of a variant strain, BI/NAP1 ribotype 027, responsible for several outbreaks and high mortality rates worldwide, the epidemiology of the CDI changed drastically in the United States, Europe, and some Latin American countries. Although the epidemic strain 027 was not yet detected in Brazil, there are ribotypes exclusively found in the country, such as, 131, 132, 133, 135, 142 and 143, which are responsible for outbreaks in Brazilian hospitals and nursing homes. Although PCR-ribotyping is the most used method in epidemiology studies of C. difficile, it is not available in Brazil. This study aimed to develop and validate an in-house database for detecting C. difficile ribotypes, usually involved in CDI in Brazilian hospitals, by using MALDI-TOF MS. A database with 19 different ribotypes, 13 with worldwide circulation and 6 Brazilian-restricted, was created based on 27 spectra readings of each ribotype. After BioNumerics analysis, neighbor-joining trees revealed that spectra were distributed in clusters according to ribotypes, showing that MALDI-TOF MS could discriminate all 19 ribotypes. Moreover, each ribotype showed a different profile with 42 biomarkers detected in total. Based on their intensity and occurrence, 13 biomarkers were chosen to compose ribotype-specific profiles, and in silico analysis showed that most of these biomarkers were uncharacterized proteins or well-conserved peptides, such as ribosomal proteins. A double-blind assessment using the 13 biomarkers correctly assigned the ribotype in 73% of the spectra analyzed, with 94%-100% of correct hits for 027 and for Brazilian ribotypes. Although further analyses are required, our results show that MALDI-TOF MS might be a reliable, fast and feasible alternative for epidemiological surveillance of C. difficile in Brazil.

Biofilm production and distribution of pilus variants among Streptococcus agalactiae isolated from human and animal sources.

Streptococcus agalactiae (group B Streptococcus, GBS) is a major pathogen in humans and animals. Pili and biofilm may be important virulence factors in this bacterial species. Here, biofilm production and the distribution of pilus variants among 134 GBS isolates from human and animal sources were evaluated. Biofilm production was significantly enhanced in 1% glucose-supplemented medium (p < 0.05). Using this medium, most GBS strains were strong biofilm producers. Biomass was mainly composed of proteins, followed by extracellular DNA, while polysaccharides represented a minor portion. All GBS strains presented at least one pilus variant. PI-2a was the most common among human GBS while PI-2b was the most common among animal isolates. Human GBS harboring PI-2b and animal GBS harboring PI-2a presented significantly reduced biofilm production (p = 0.0033). In conclusion, strong biofilm production seems to be a common characteristic in GBS, and association of the clinical source with the pilus variant may be crucial for this.

The Convergent Effect of International Collaboration between Young Leaders of Two Global Societies: Strengthening Microbiology Education and Training Practices Worldwide.

There has never been a more critical time than the present for young microbiologists to engage in continuous postgraduate education. Next-generation microbiologists need to integrate several scientific disciplines in the early stages of their careers to keep up with the ever-changing nature of microbiology (advent of high-throughput sequencing platforms, bioinformatics, complex omic, culture and systems-biology approaches) and to ensure timely response to the current global health and biodiversity threats. Here we propose that international collaboration of young experts would help to address the challenges in microbiology training and serve as a communication bridge between senior and early career scientists. To establish a one-of-a-kind collaboration, the Trainee Association (TAE) of the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) and the Young Leaders Circle (YLC) of the American Society for Microbiology (ASM) have joined forces to adequately represent the interests of young members within the two largest societies in the field of microbiology. Through this partnership, both societies aim to promote continuous education for young scientists and the general public. Additionally, we intend to identify shared objectives and introduce joint activities that could lead to a beneficial exchange process for a long-lasting relationship. This partnership further contributes to advancing both the quality and quantity of education.

En la actualidad es crítico que los jóvenes microbiólogos se involucren en estudios de posgrado y formación continua. La próxima generación de microbiólogos necesita integrar varias disciplinas científicas durante las primeras etapas de sus carreras para mantenerse al día con la naturaleza siempre cambiante de la microbiología (por ejemplo, con el advenimiento de las plataformas de secuenciación masiva, de las herramientas bioinformáticas, de las técnicas ómicas avanzadas, de cultivo y de biología de sistemas), y para asegurar una respuesta oportuna ante las amenazas mundiales actuales para la salud y la biodiversidad. Se propone que la colaboración internacional de jóvenes expertos ayudará a enfrentar los desafíos de la formación en microbiología y servirá como un puente de comunicación entre científicos experimentados y en etapas tempranas de sus carreras. Para establecer una colaboración única, la Asociación de Pasantes (TAE) de la Sociedad Europea de Microbiología Clínica y Enfermedades Infecciosas (ESCMID) y el Círculo de Jóvenes Líderes (YLC) de la Sociedad Americana de Microbiología (ASM) han conjuntado esfuerzos para representar adecuadamente los intereses de los jóvenes integrantes de las dos sociedades más numerosas en la disciplina de la microbiología. A través de esta colaboración, ambas sociedades tienen como objetivo promover la formación continua de jóvenes científicos y del público en general. Además, se tiene la intención de identificar objetivos compartidos y proponer actividades conjuntas que propicien un proceso de intercambio beneficioso para mantener una relación duradera. Esta colaboración contribuye a mejorar tanto la calidad como la cantidad de la educación.

DivulgaMicro : A Brazilian Initiative To Empower Early-Career Scientists with Science Communication Skills.

Communication is fundamental in science. Among scientists, communication skills are required to write a comprehensible scientific manuscript or to prepare an attractive oral presentation. In addition, the ability to communicate successfully with the nonscientific community has been increasingly appreciated, as it represents the most effective way to promote popular scientific literacy. Nevertheless, students majoring in sciencerelated courses are not trained specifically for these purposes, and improving communication skills usually depends on tips and advice given by peers. To this end, we have launched DivulgaMicro, an initiative that aims to enhance science communication among early-career scientists in Brazil. DivulgaMicro relies on two major cornerstones: providing online resources via a dedicated website (www.divulgamicro.com.br/index.html) and promoting onsite workshops in universities located in different macro regions of the country. On the website, people can access a collection of fun activities designed to teach scientific concepts to a general audience, along with tips and news regarding public outreach events in Brazil. In the month following its launch, our website had 1,026 visitors from 10 different countries besides Brazil. Regarding the workshops, six were offered during 2018, with nearly 600 attendees. In the course, scientists are presented best practices for scientific writing and oral presentations, as well as techniques to improve communication with lay audiences, such as the use of storytelling structure and analogies. There is a high demand for science communication resources in Brazil, attesting to the importance of initiatives such as DivulgaMicro in our country.

A comunicação é fundamental para a ciência e a comunidade científica, cuja habilidade em se comunicar é necessária tanto para a escrita de um artigo científico quanto para a preparação de uma apresentação oral atrativa. Além disso, a capacidade de se comunicar de forma bem-sucedida com o público leigo tem sido cada vez mais apreciada, uma vez que representa a maneira mais eficaz em promover a alfabetização científica da população. No entanto, os estudantes que buscam se engajar na carreira científica não recebem treinamento específico nesse sentido, e o desenvolvimento dessa habilidade geralmente é dependente de dicas e conselhos recebidos de seus pares. Nesse contexto, foi criado o projeto DivulgaMicro, uma iniciativa que visa promover o desenvolvimento da comunicação científica entre jovens cientistas no Brasil. O projeto DivulgaMicro possui dois pilares principais: oferecer recursos online através do website http://www.divulgamicro.com.br/index.html e promover workshops de Comunicação e Divulgação Científica em universidades de diferentes macrorregiões do país. No website, as pessoas podem acessar e baixar uma coleção de atividades lúdicas destinadas ao ensino de conceitos científicos para o público em geral, incluindo crianças e adolescentes, além de dicas e novidades sobre divulgação científica no país. Após um mês de lançamento do projeto, o website do DivulgaMicro já havia recebido 1.026 acessos de 10 diferentes países além do Brasil. Em relação aos workshops, estão previstas seis edições em 2018 alcançando um público de cerca de 800 pessoas. No workshop, o participante é apresentado às melhores e mais eficazes práticas de escrita científica e apresentação oral, bem como técnicas para desenvolver e aprimorar a comunicação com o público leigo, incluindo como utilizar a estrutura storytelling e a aplicação de analogias. Atualmente no Brasil existe uma alta demanda em recursos e fontes de aprendizado em comunicação e divulgação científica, o que corrobora a relevância de iniciativas como o DivulgaMicro em nosso país.

Evolution of Penicillin Non-susceptibility Among Streptococcus pneumoniae Isolates Recovered From Asymptomatic Carriage and Invasive Disease Over 25 years in Brazil, 1990-2014.

Streptococcus pneumoniae is a major cause of community-acquired pneumonia and meningitis, and it is also found as a commensal, colonizing the human upper respiratory tract of a portion of the human population. Its polysaccharide capsule allows the recognition of more than 90 capsular types and represents the target of the currently available pneumococcal conjugate vaccines (PCVs), such as the 10-valent (PCV10) and the 13-valent (PCV13). Penicillin non-susceptible pneumococci (PNSP) have been listed as one of the current major antimicrobial-resistant pathogen threats. In Brazil, the emergence of PNSP was initially detected in the mid 1990s and PCV10 has been part of the National Immunization Program since 2010. Here, we investigated the distribution of capsular types and penicillin susceptibility profiles of 783 pneumococcal strains isolated in Brazil between 1990 and 2014 to assess the evolution of penicillin non-susceptibility among pneumococci associated with asymptomatic carriage and invasive pneumococcal disease (IPD). The most common serotypes among carriage isolates were 19F, 6B, 6C, 23F, and 14. Among IPD isolates, the most frequent types were 14, 3, 6B, 5, 19F, and 4. We detected 21 types exclusively associated with IPD isolates, whereas non-typeable (NT) isolates were only detected in carriage. Nearly half of the isolates belonged to PCV10 serotypes, which remarkably decreased in occurrence (by nearly 50%) after PCV10 introduction (2011-2014), while non-PCV10 serotypes increased. PNSP frequency and levels were much higher among carriage isolates, but PNSP belonging to PCV10 serotypes were more common in IPD. While the occurrence of PNSP has decreased significantly among IPD isolates since 2011, it kept increasing among carriage strains. Such a difference can be attributed to the serotypes that emerged in each clinical source after PCV10 usage. PNSP with multidrug resistance profiles that emerged within carriage isolates comprised mostly serotypes 6C and 35B, as well as NT isolates. In turn, penicillin-susceptible capsular types 3, 20, and 8 have risen among IPD. Overall, our results reinforce the relevance of PNSP surveillance over a long period of time to better understand the dynamics of antimicrobial resistance in response to PCV introduction and may also contribute to improve control measures toward drug-resistant pneumococci.

Conjugative transfer of resistance determinants among human and bovine Streptococcus agalactiae.

Streptococcus agalactiae (GBS) is a major source of human perinatal diseases and bovine mastitis. Erythromycin (Ery) and tetracycline (Tet) are usually employed for preventing human and bovine infections although resistance to such agents has become common among GBS strains. Ery and Tet resistance genes are usually carried by conjugative transposons (CTns) belonging to the Tn916 family, but their presence and transferability among GBS strains have not been totally explored. Here we evaluated the presence of Tet resistance genes (tetM and tetO) and CTns among Ery-resistant (Ery-R) and Ery-susceptible (Ery-S) GBS strains isolated from human and bovine sources; and analyzed the ability for transferring resistance determinants between strains from both origins. Tet resistance and int-Tn genes were more common among Ery-R when compared to Ery-S isolates. Conjugative transfer of all resistance genes detected among the GBS strains included in this study (ermA, ermB, mef, tetM and tetO), in frequencies between 1.10(-7) and 9.10(-7), was possible from bovine donor strains to human recipient strain, but not the other way around. This is, to our knowledge, the first report of in vitro conjugation of Ery and Tet resistance genes among GBS strains recovered from different hosts.

Conjugative transfer of resistance determinants among human and bovine Streptococcus agalactiae

Streptococcus agalactiae (GBS) is a major source of human perinatal diseases and bovine mastitis. Erythromycin (Ery) and tetracycline (Tet) are usually employed for preventing human and bovine infections although resistance to such agents has become common among GBS strains. Ery and Tet resistance genes are usually carried by conjugative transposons (CTns) belonging to the Tn916 family, but their presence and transferability among GBS strains have not been totally explored. Here we evaluated the presence of Tet resistance genes (tetM and tetO) and CTns among Ery-resistant (Ery-R) and Ery-susceptible (Ery-S) GBS strains isolated from human and bovine sources; and analyzed the ability for transferring resistance determinants between strains from both origins. Tet resistance and int-Tn genes were more common among Ery-R when compared to Ery-S isolates. Conjugative transfer of all resistance genes detected among the GBS strains included in this study (ermA, ermB, mef, tetM and tetO), in frequencies between 1.10-7 and 9.10-7, was possible from bovine donor strains to human recipient strain, but not the other way around. This is, to our knowledge, the first report of in vitro conjugation of Ery and Tet resistance genes among GBS strains recovered from different hosts.

Nasopharyngeal carriage, serotype distribution and antimicrobial resistance of Streptococcus pneumoniae among children from Brazil before the introduction of the 10-valent conjugate vaccine.

BACKGROUND: Streptococcus pneumoniae remains a major cause of childhood morbidity and mortality worldwide. Nasopharyngeal colonization plays an important role in the development and transmission of pneumococcal diseases, and infants and young children are considered to be the main reservoir of this pathogen. The aim of this study was to evaluate the rates and characteristics associated with nasopharyngeal carriage, the distribution of serotypes and the antimicrobial resistance profiles of Streptococcus pneumoniae among children in a large metropolitan area in Brazil before the introduction of the 10-valent pneumococcal conjugate vaccine. METHODS: Between March and June 2010, nasopharyngeal swabs were collected from 242 children aged <6 years attending one day care center and the emergency room of a pediatric hospital. Pneumococcal isolates were identified by conventional methods and serotypes were determined by a sequential multiplex PCR assay and/or the Quellung reaction. The antimicrobial susceptibilities of the pneumococci were assessed by the disk diffusion method. MICs for erythromycin and penicillin were also performed. Erythromycin resistance genes were investigated by PCR. RESULTS: The overall colonization rate was 49.2% and it was considerably higher among children in the day care center. Pneumococcal carriage was more common among day care attenders and cohabitants with young siblings. The most prevalent serotypes were 6B, 19F, 6A, 14, 15C and 23F, which accounted for 61.2% of the isolates. All isolates were susceptible to clindamycin, levofloxacin, rifampicin and vancomycin. The highest rate of non-susceptibility was observed for sulphamethoxazole-trimethoprim (51.2%). Penicillin non-susceptible pneumococci (PNSP) accounted for 27.3% of the isolates (MICs of 0.12-4 µg/ml). Penicillin non-susceptibility was strongly associated with serotypes 14 and 23F. Hospital attendance and the presence of respiratory or general symptoms were frequently associated with PNSP carriage. The two erythromycin-resistant isolates (MICs of 2 and 4 µg/ml) belonged to serotype 6A, presented the M phenotype and harbored the mef(A/E) gene. CONCLUSIONS: Correlations between serotypes, settings and penicillin non-susceptibility were observed. Serotypes coverage projected for the 10-valent pneumococcal conjugate vaccine was low (45.5%), but pointed out the potential reduction of PNSP nasopharyngeal colonization by nearly 20%.