Francisella salimarina infection in farmed cobia (Rachycentron canadum) and dusky grouper (Epinephelus marginatus) in Brazil: A case report.

This study aimed to identify and characterize isolates of Francisella salimarina associated with an outbreak on a marine fish farm in Brazil and to analyse their genetic variability and antimicrobial susceptibility. In 2021, diseased cobias (Rachycentron canadum, n = 10) and dusky groupers (Epinephelus marginatus, n = 10) were sampled and subjected to bacteriological and pathological examinations. The isolates obtained were morphologically and biochemically characterized and identified by matrix-assisted laser desorption/ionization-time of flight (MALDI-ToF) and 16S rRNA gene sequencing. The genetic diversity of these isolates was analysed using repetitive extragenic palindromic-polymerase chain reaction (REP-PCR). Antimicrobial susceptibility was assessed using the disk diffusion technique. Macroscopically, the fish presented skin ulcerations, ocular lesions, hepatomegaly and splenomegaly. A pleomorphic, gram-negative, catalase- and oxidase-positive bacterium was isolated from seven cobias and two groupers. The 16S rRNA gene sequences showed >99% coverage and identity with other deposited sequences of F. salimarina. The results of the biochemical analysis corresponded to these bacterial species. Histologically, granulomas were observed in the spleen, liver and heart of the cobias (n = 6), and necrotizing and fibrinous dermatitis and myositis were identified in some groupers (n = 2). The isolates exhibited the same banding pattern when REP-PCR was performed, indicating that they were clonally related. Finally, the antibiogram test, no inhibition halo was observed for amoxicillin and trimethoprim-sulfamethoxazole. To our knowledge, this is the first report of F. salimarina infection in cobias and dusky groupers.

Edwardsiella tarda in Tambaqui (Colossoma macropomum): A Pathogenicity, Antimicrobial Susceptibility, and Genetic Analysis of Brazilian Isolates.

Edwardsiella tarda is a crucial pathogenic bacterium in tropical aquaculture. This bacterium was recently isolated from tambaqui (Colossoma macropomum), a commercially important fish species in Brazil. This study assessed the antimicrobial susceptibility, pathogenicity, and genetic diversity of the tambaqui-derived E. tarda isolates. Fourteen bacterial isolates isolated from tambaqui were identified as E. tarda by using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry and dnaJ gene sequencing. Antimicrobial susceptibility tests were conducted against seven drugs using the disc diffusion assay. The pathogenicity test conducted by intraperitoneal injection of 2.4 × 107 colony-forming units (CFU) fish-1 of E. tarda (ED38-17) into tambaqui juveniles eventually revealed that neither clinical signs nor death were present. However, splenomegaly and whitish areas in the spleen and kidneys were observed. The histological investigation also revealed granulomatous splenitis, nephritis, and hepatitis occurring internally. Repetitive extragenic palindromic-PCR fingerprinting separated the 14 isolates into three genetic groups. The antibiogram revealed that all E. tarda isolates were wild-type (WT) to florfenicol (FLO), norfloxacin (NOR), neomycin (NEO), erythromycin (ERY), and oxytetracycline (OXY); however, some were non-wild-type to sulfamethoxazole/trimethoprim (7.1%) and amoxicillin (21.4%). Therefore, through experimental infection, E. tarda ED38-17 could induce pathogenic effects in C. macropomum. Additionally, three distinct genetic types were found, and the E. tarda isolates were WT to FLO, NOR, NEO, ERY, and OXY. These findings raise awareness of a bacteria causing unseen lesions, a pathogen that will potentially impact tambaqui aquaculture in the future.

Effective Biocorrosive Control in Oil Industry Facilities: 16S rRNA Gene Metabarcoding for Monitoring Microbial Communities in Produced Water.

Microbiologically influenced corrosion (MIC) or biocorrosion is a complex biological and physicochemical process, Strategies for monitoring MIC are frequently based on microbial cultivation methods, while microbiological molecular methods (MMM) are not well-established in the oil industry in Brazil. Thus, there is a high demand for the development of effective protocols for monitoring biocorrosion with MMM. The main aim of our study was to analyze the physico-chemi- cal features of microbial communities occurring in produced water (PW) and in enrichment cultures in oil pipelines of the petroleum industry. In order to obtain strictly comparable results, the same samples were used for both culturing and metabarcoding. PW samples displayed higher phylogenetic diversity of bacteria and archaea whereas PW enrichments cultures showed higher dominance of bacterial MIC-associated genera. All samples had a core community composed of 19 distinct genera, with MIC-associated Desulfovibrio as the dominant genus. We observed significant associations between the PW and cultured PW samples, with a greater number of associations found between the cultured sulfate-reducing bacteria (SRB) samples and the uncultured PW samples. When evaluating the correlation between the physicochemical characteristics of the environment and the microbiota of the uncultivated samples, we suggest that the occurrence of anaerobic digestion metabolism can be characterized by well-defined phases. Therefore, the detection of microorganisms in uncultured PW by metabarcoding, along with physi-cochemical characterization, can be a more efficient method compared to the culturing method, as it is a less laborious and cost-effective method for monitoring MIC microbial agents in oil industry facilities.

Cost analysis of motorcycle accident victims at a university hospital: perspectives from 2017 and 2020 / Análise de custo das vítimas de acidente motociclístico em um hospital universitário: perspectivas de 2017 e 2020

ABSTRACT Introduction: Motorcycle accidents constitute a public health problem that affects public and private health services due to the expenses of the victim's treatment and rehabilitation. Objective: Evaluate the impact of motorcycle accident costs in a university hospital in 2020. Method: Comparative analysis of the costs of motorcycle accident patients in 2020 and 2017. Results: Among 151 patients included in the study, the average cost was U$3,083.54, and the average days of hospitalization were 5.3 days. The patient with the highest cost to the hospital spent U$22,504.05, and the patient with the lowest cost spent U$356.72. The longest stay among these patients was 41 days, and the shortest was one day. The average cost per patient per day for the entire sample was U$581.80. Conclusion: The formulation and application of strategies that promote the reduction of motorcycle accidents in the city of Campinas are necessary. Level of evidence II, Retrospective study.

RESUMO Introdução: Acidentes motociclísticos constituem um problema de saúde pública que atinge os serviços públicos e privados de saúde, em função dos gastos com o tratamento e com o processo de reabilitação da vítima. Objetivo: Avaliar o impacto dos custos dos acidentes motociclísticos em um hospital universitário em 2020. Método: Análise comparativa dos custos dos pacientes vítimas de acidente motociclístico no ano de 2020 e 2017. Resultados: Dentre 151 pacientes incluídos no estudo, o custo médio foi de U$3.083,54 e a média de dias de internação foi de 5,3 dias. O paciente que apresentou maior custo para o hospital, teve um gasto de U$22.504,05 e o que teve o menor custo, gastou U$356,72. O maior tempo de internação, entre estes pacientes, foi de 41 dias e o menor tempo foi de 1 dia. O custo médio por paciente por dia, em toda a amostra, foi de U$581,80. Conclusão: Faz-se necessário a formulação e aplicação de estratégias que promovam a redução dos acidentes motociclísticos na cidade de Campinas. Nível de evidência II; Estudo retrospectivo.

EPIDEMIOLOGICAL DISTRIBUTION OF SOFT PART TUMORS IN A TERTIARY HOSPITAL.

Objective: To evaluate cases of soft tissue tumors at the Orthopedic Oncology service of the PUC-Campinas Hospital and determine the epidemiological profile in the period from February 2012 to November 2019, associating the participation of a non-reference hospital in the approach and treatment of the pathology. Methods: This study evaluated 72 patients aged 18 to 81 years, of both sexes and diagnosed with soft tissue tumor and divided into two groups: (I) primary etiology and (II) metastatic etiology. Results: Of 146 patients admitted, 22 resulted in deaths, with 9 patients from Group I, and 13 from Group II. For all patients with soft tissue tumor, aged between 51 and 58 years, admitted in the period, the probability of survival after 46 months was 71.84%, Group I's was 22.7% and Group II's 91.43%. Conclusion: Despite the scarcity of epidemiological data related to soft tissue tumor, the data appreciated in the hospital's service, not a reference in treating this type of condition, are compatible with the data presented in specialized hospitals in Brazil, thus, in confluence with the literature. Level of Evidence III, Comparative, Prognostic and Retrospective Study.

Objetivo: Avaliar casos de tumores de partes moles do Serviço de Oncologia Ortopédica do Hospital PUC-Campinas e determinar o perfil epidemiológico no período de fevereiro de 2012 a novembro de 2019, associando à participação de um hospital que não é referência na abordagem e tratamento da patologia. Métodos: Foi realizada uma análise exploratória de dados com 72 pacientes entre 18 e 81 anos, de ambos os sexos e com diagnóstico de tumor de partes moles. Eles foram avaliados e divididos em dois grupos: (I) de etiologia primária e (II) de etiologia metastática. Resultados: Do total de 146 pacientes admitidos, 22 vieram a óbito, sendo 9 pacientes do Grupo I, e 13 do Grupo II. Para todos os pacientes com tumor de partes moles, com idade entre 51 e 58 anos, admitidos no período, a probabilidade de sobrevida após 46 meses foi de 71,84%, enquanto do Grupo I foi de 22,7%, e do Grupo II foi de 91,43%. Conclusão: Apesar da escassez de dados epidemiológicos relacionados ao tumor de partes moles, os dados apreciados no serviço do hospital, que não é referência no tratamento dessa condição, são compatíveis com os dados apresentados em hospitais especializados no Brasil, assim, corroborando a literatura. Nível de Evidência III, Estudo Retrospectivo Comparativo Prognóstico.

Comparative polyphasic characterization of Weissella strains isolated from beaked whale and rainbow trout (Oncorhynchus mykiss): confirmation of Weissella ceti sp. nov. and description of the novel Weissella tructae sp. nov. isolated from farmed rainbow trout.

The weissellosis agent bacterium (WS08T = CBMAI 2730) was isolated from diseased rainbow trout (Oncorhynchus mykiss) in Brazil. The whole genome sequence of this strain was compared with the Mexican W-1 strain, also isolated from diseased rainbow trout, and with the Weissella ceti type strain CECT 7719 T (= 1119-1A-09 T = CCUG 59653 T), recovered from the beaked whale. Digital DNA-DNA hybridization pairwise analyses scored 98.7% between the Mexican W-1 and Brazilian WS08T but just 24.4% for both fish isolates compared to the W. ceti type strain CECT 7719 T. The 16S rRNA gene sequence comparisons with isolates of W. ceti, available at GenBank, were conducted. All rainbow trout-pathogenic isolates grouped close (97% bootstrap confirmation), but when this group was compared to the W. ceti type strain CECT 7719 T the similarity varied from 78.9 to 79.1%. Phenotypic assays were also conducted, and the W. ceti type strain diverged from WS08T and W-1 in the hydrolysis of aesculin, D-mannose, and potassium gluconate and in the hydrolysis of hippurate. Moreover, WS08T and W-1 showed weak growth at 5 °C whereas no growth was observed for W. ceti CECT 7719 T. The major fatty acids (> 10% total fatty acids) presented by WS08T and W-1 were summed feature 8 (C18:1 ω7c/C18:1 ω6c), summed feature 3 (C16:1 ω6c/C16:1ω7c), and C16:0. The results of phylogenetic and phenotypic analyses clearly differentiated the W. ceti CECT 7719 T type strain from the assessed pathogenic strains obtained from rainbow trout. Therefore, Weissella strains isolated from rainbow trout, here represented by strain WS08T (= CBMAI 2730), should be known as members of a novel species for which the name Weissella tructae sp. nov. is proposed.

The impact of covid-19 on the profile of motorcycle accidents attended at a tertiary hospital in campinas / Impacto da covid-19 no perfil de acidentados de moto atendidos em um hospital terciário em campinas

ABSTRACT Traffic-accidents are a public health problem with repercussions on population morbimortality. Objective: To analyze the impact of the pandemic on the profile of motorcycle accidents assisted at the Tertiary Hospital in 2020. Methods: Cross-sectional, descriptive retrospective study in 2017 and 2020 of 260 medical records of care for motorcycle accidents in the emergency room of the Tertiary Hospital. Statistical analysis of data and their correlations using the chi-square test (p < 0.05). Results: Of the 105 medical records in 2017, 83% are men, mean age 29.8 years, and death rate of 3.90%. Fractures in 98.10%, 64.10% exposed and predominantly the tibia (61.90%). Of the 155 medical records in 2020, 91.61% are men, mean age 31.21 years, and no deaths. Fractures in 94.84%, 37.42% exposed and predominantly the tibia (28.57%). Between 2017 and 2020, Infosiga-SP showed a relevant reduction (p < 0.001) of deaths in the hospital environment (52.46% to 31.91%). Conclusion: The incidence of motorcycle accidents increased, in-hospital deaths dropped, but the epidemiological profile of accidents at the Hospital remained unchanged. Level of Evidence III, Comparative Retrospective Study.

RESUMO Acidentes motociclísticos configuram um problema de saúde pública com repercussões na morbimortalidade populacional. Objetivo: Analisar o impacto da pandemia por COVID-19 no perfil de acidentes motociclísticos atendidos em um hospital terciário em 2020. Métodos: Estudo transversal, descritivo e retrospectivo que analisou 260 prontuários de acidentados de moto atendidos na Urgência e Emergência do Hospital Universitário Terciário em 2017 e 2020. Realizou-se uma análise estatística dos dados e suas correlações pelo teste qui-quadrado (p < 0,05). Resultados: Dos 105 prontuários de 2017, 83% são de homens (p < 0,001), com média de 29,8 anos, e índice de óbito de 3,90%. Houve fraturas em 98,10% dos casos, sendo 58,10% expostas e predominantemente da tíbia (61,90%). Dos 155 prontuários de 2020, 91,61% são homens (p < 0,001), com média de 31,21 anos e sem casos de óbito. Houve fraturas em 94,84%, sendo 37,42% expostas e predominantemente da tíbia (28,57%). Entre 2017 e 2020, o Infosiga-SP mostrou redução significativa (p < 0,001) de mortes em ambiente hospitalar (de 52,46% para 31,91%). Conclusão: Houve aumento na incidência dos acidentes motociclísticos e queda nos óbitos intra-hospitalares, mas o perfil epidemiológico dos acidentados no hospital permaneceu inalterado. Nível de Evidência III, Estudo Comparativo Retrospectivo.

Epidemiological distribution of soft part tumors in a tertiary hospital / Distribuição epidemiológica de tumores de partes moles em um hospital terciário

ABSTRACT Objective: To evaluate cases of soft tissue tumors at the Orthopedic Oncology service of the PUC-Campinas Hospital and determine the epidemiological profile in the period from February 2012 to November 2019, associating the participation of a non-reference hospital in the approach and treatment of the pathology. Methods: This study evaluated 72 patients aged 18 to 81 years, of both sexes and diagnosed with soft tissue tumor and divided into two groups: (I) primary etiology and (II) metastatic etiology. Results: Of 146 patients admitted, 22 resulted in deaths, with 9 patients from Group I, and 13 from Group II. For all patients with soft tissue tumor, aged between 51 and 58 years, admitted in the period, the probability of survival after 46 months was 71.84%, Group I's was 22.7% and Group II's 91.43%. Conclusion: Despite the scarcity of epidemiological data related to soft tissue tumor, the data appreciated in the hospital's service, not a reference in treating this type of condition, are compatible with the data presented in specialized hospitals in Brazil, thus, in confluence with the literature. Level of Evidence III, Comparative, Prognostic and Retrospective Study.

RESUMO Objetivo: Avaliar casos de tumores de partes moles do Serviço de Oncologia Ortopédica do Hospital PUC-Campinas e determinar o perfil epidemiológico no período de fevereiro de 2012 a novembro de 2019, associando à participação de um hospital que não é referência na abordagem e tratamento da patologia. Métodos: Foi realizada uma análise exploratória de dados com 72 pacientes entre 18 e 81 anos, de ambos os sexos e com diagnóstico de tumor de partes moles. Eles foram avaliados e divididos em dois grupos: (I) de etiologia primária e (II) de etiologia metastática Resultados: Do total de 146 pacientes admitidos, 22 vieram a óbito, sendo 9 pacientes do Grupo I, e 13 do Grupo II. Para todos os pacientes com tumor de partes moles, com idade entre 51 e 58 anos, admitidos no período, a probabilidade de sobrevida após 46 meses foi de 71,84%, enquanto do Grupo I foi de 22,7%, e do Grupo II foi de 91,43%. Conclusão: Apesar da escassez de dados epidemiológicos relacionados ao tumor de partes moles, os dados apreciados no serviço do hospital, que não é referência no tratamento dessa condição, são compatíveis com os dados apresentados em hospitais especializados no Brasil, assim, corroborando a literatura. Nível de Evidência III, Estudo Retrospectivo Comparativo Prognóstico.

The impact of motorcycle accidents in a reference hospital during the covid-19 pandemic / Impacto nos acidentes motociclísticos em um hospital referência durante a pandemia de covid-19

ABSTRACT Objective: This study aimed to analyze the relationship between the number of motorcycle accidents attended at a reference Hospital for trauma in Campinas, state of São Paulo, and the pandemic by COVID-19 during the year 2020. Methods: This is a cross-sectional, descriptive retrospective study carried out at Hospital PUC- Campinas, through the analysis of medical records of patients, victims of motorcycle trauma undergoing orthopedic surgical management in 2020. The phases of the pandemic and the isolation rates according to the São Paulo Plan were studied. Chi-Square tests, and the Least Squares method were applied for statistical calculations. Results: 155 medical records were analyzed, of which 91.61% of the patients were male. Of those admitted, 94.84% suffered fractures and 51.61%, polyfractures. There was a correlation between the average isolation rates and the number of accidents. In the 14-day period, as the average isolation rate increased by 10%, there was an increase of approximately 3 accidents in that period. In the 7-day analysis, 1.7 more traumas were observed for every 10% increase in the average isolation. Conclusion: The results suggest that by increasing the average isolation rate, the number of traumas per motorcycle treated at the institution increased. Level of Evidence III, Comparative Retrospective Study.

RESUMO Objetivos: Analisar se houve relação do número de acidentes motociclísticos atendidos em um hospital referência para o tratamento de trauma em Campinas com a pandemia pela COVID-19 durante o ano de 2020. Métodos: Estudo transversal, retrospectivo descritivo, realizado no Hospital PUC-Campinas, por meio da análise de prontuários de vítimas de trauma por acidente de moto com conduta cirúrgica ortopédica em 2020. Foram estudadas as fases da pandemia e as taxas de isolamento de acordo com o Plano São Paulo (Plano SP). Utilizaram-se análises de p-valor usando testes qui-quadrado e o método de mínimos quadrados para cálculos estatísticos. Resultados: Foram analisados 155 prontuários de pacientes, dos quais 91,61% eram do sexo masculino. Dos internados, 94,84% sofreram fraturas e 51,61%, polifraturas. Houve correlação entre as taxas médias de isolamento com o número de acidentes. No período de 14 dias, à medida em que houve crescimento de 10% na média da taxa de isolamento, observou-se um aumento de aproximadamente três acidentes. Na análise de 7 dias, observou-se 1,7 traumas a mais a cada 10% do aumento da média de isolamento. Conclusão: Os resultados sugerem que, ao aumentar a taxa média de isolamento, houve um aumento do número de traumas por acidente de moto atendidos na instituição. Nível de Evidência III, Estudo Retrospectivo Comparativo.

The importance of the therapeutic care plan in colorectal surgery in a teaching hospital

Introduction: A therapeutic plan is elaborated based on the health needs of each user, allowing a multidisciplinary team to assess diagnoses, treatment options, bonds, and optimal hospitalization time. Objectives: To identify risk management tools already used and implemented in a reference teaching hospital in the city of São Paulo and to analyze their application and risk factors in medium and large colorectal surgery. Method: Observational, longitudinal, and prospective study, with 30 patients with colorectal disease hospitalized in the surgical ward of the coloproctology service and in need of surgical treatment. In the first group, the protocol was applied with the knowledge of the researcher only, and, in the second group, with the knowledge of both the researcher and the attending physicians. Results: Sixty percent of the patients were female with a mean age of 60.93 years and body mass index (BMI) of 26.07 Kg/m2. After surgery, patients in the first group who did not receive venous thromboembolism (VTE) prophylaxis in the first 24 hours had an increased risk of having the event compared with those who returned to prophylaxis (p<0.005), thus suggesting this prophylaxis was a protective factor against thromboembolic event (p=0.006). This group also had a higher risk of hypoglycemia when no strict control was performed (p=0.041). Conclusion: The compliance to hospital protocols with applicationmonitoring, notedly in teaching places with annual admission of resident physicians, is a fundamental part of the adequate care of the patient combined with the implementation of therapeutic plans. (AU)

Study of defecation disorders in elderly patients / Estudo dos distúrbios da defecação em pacientes idosos

Abstract Introduction: Defecation disorders, whether anal incontinence or chronic intestinal constipation, are frequent pelvic floor alterations in the general population and are more common in those with risk factors,i.e., in the elderly, women with an obstetric background, and those with comorbidities, history of pelvic radiotherapy, diabetics, the bedridden, or those with history of orifice surgery, among others. Objective: To analyze the incidence of defecation disorders in geriatric patients treated at the Medical Specialties Outpatient Service (MSOS) of Hospital Santa Marcelina. Methods: Prospective, randomized study that interviewed the same patients in two moments: 1) subjective anamnesis through spontaneous history and 2) objective anamnesis with specific questionnaires to assess anal incontinence and chronic constipation. Results: Between March 2016 and June 2017, 149 patients were analyzed, of whom 114 (76.5%) were female, with a similar mean age between genders; 51.67% had symptoms of anal incontinence and/or chronic constipation. Only 35.5% of patients with complaints of fecal leakage or flatus spontaneously reported them, while 87.1% of constipated patients did so. In the present study, no significant correlation was observed between the mode of delivery (p = 0.106), pregnancy (p = 0.099), and the number of deliveries (p = 0.126) with anal incontinence. In turn, there was no higher incidence of chronic intestinal constipation in females (p = 0.099) and most patients with this complaint had Bristol type 1 or 2 stools. Conclusion: The incidence of defecation disorders in the geriatric population is high and, most notably, anal incontinence is not spontaneously reported by most patients.

Resumo Introdução: Os distúrbios da evacuação, seja a incontinência anal ou a constipação intestinal crônica, representam alterações do assoalho pélvico bastante frequente na população em geral e mais comumente naqueles com fatores de risco, ou seja, em idosos, mulheres com passado obstétrico, comorbidades, antecedente de radioterapia pélvica, diabéticos, acamados, história de cirurgias orificiais, dentre outros. Objetivo: Analisar a incidência de distúrbios defecatórios em pacientes geriátricos atendidos no Ambulatório de Especialidades Médicas (AME) do Hospital Santa Marcelina. Metodologia: Estudo prospectivo e aleatório com a entrevista do mesmo paciente em dois momentos: 1) Anamnese subjetiva através da história espontânea e 2) Anamnese objetiva com questionários específicos para avaliação de incontinência anal e constipação intestinal crônica. Resultados: Foram analisados 149 pacientes entre Março de 2016 e Junho de 2017, sendo 114 (76,5%) do sexo feminino com média de idade semelhante entre os sexos; 51,67% apresentavam sintomas de incontinência anal e/ou constipação intestinal crônica. Apenas 35,5% dos pacientes com queixas de escape de fezes ou flatos relataram de forma espontânea e 87,1% dos pacientes constipados o fizeram. No presente estudo não se verificou correlação significativa entre via de parto p = 0,106, gestação p = 0,099 e número de partos p = 0,126 com incontinência anal. Por outro lado, não se verificou maior incidência de constipação intestinal crônica no sexo feminino p = 0,099 e a maioria dos pacientes com essa queixa apresentavam fezes ressecadas tipo Bristol 1 ou 2. Conclusão: Incidência de distúrbios da defecação na população geriátrica é elevada e, notadamente a IA não é referida de forma espontânea pela maioria dos pacientes.

Cell wall glycolipids from Corynebacterium pseudotuberculosis strains with different virulences differ in terms of composition and immune recognition.

Caseous lymphadenitis (CLA) is an infectious disease caused by Corynebacterium pseudotuberculosis in small ruminants and is characterized by the development of granulomas in the lymph nodes, spleen, liver, and lungs. Although little is known about the host-pathogen relationship of this bacterium, it was previously reported that the pathogen's lipids are important for its taxonomic classification and survival inside macrophages. However, there are no studies regarding the composition of these molecules. In this study, cell wall glycolipids from two C. pseudotuberculosis strains presenting different virulence profiles were purified and its composition was characterized. A difference was observed between the electrophoretic and chromatogram profiles for cell wall components from the two strains, mainly among molecules with low molecular weights. IgM from sheep with acute CLA recognized antigens with an estimated molecular weight of 11 kDa of the low-pathogenicity strain, while low-molecular weight antigens from the high-pathogenicity strain presented a lower recognition by these antibodies. Mass spectrometry analysis showed that the cell wall of the high-pathogenicity strain contained glycolipids with high amounts of unsaturated fatty acids and glycerophosphoinositols, which may contribute to the capacity of this strain to cause severe disease. In conclusion, it is indicated that cell wall non-protein antigens can play a key role in C. pseudotuberculosis virulence.

Atypical Multibacterial Granulomatous Myositis in a Horse: First Report in Italy.

Infectious causes of myositis are reported relatively uncommonly in horses. Among them, bacterial causes include Streptococcus equi subsp. zooepidemicus, Actinobacillus equuli, Fusobacterium spp. Staphylococcus spp, and Corynebacterium pseudotuberculosis. Infection can be spread to muscles via haematogenous or extension from skin lesions. Parasitic myositis has also been documented. In this report, a 12 year-old Italian Quarter Horse mare presented with diffuse subcutaneous nodules and masses ranging from 2 × 3 to 5 × 20 cm in size, and adherent to subcutis and muscles that were first macroscopically and cytologically diagnosed as pyogranulomas. Subsequently, histological, molecular, bacteriological, and biochemical investigations were performed. All the data obtained allowed to diagnose a severe and diffuse multibacterial granulomatous myositis caused by Corynebacterium pseudotuberculosis and Corynebacterium amycolatum. Following the therapy and an initial disappearance of most of the lesions together with a general improvement of the mare, the clinical condition deteriorated, and new nodules appeared. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and PCR techniques revealed the presence of bacteria as Glutamicibacter creatinolyticus and Dietzia spp. To the authors' knowledge, this case report represents the first description of multibacterial granulomatous myositis due to Corynebacterium pseudotuberculosis, Corynebacterium amycolatum, Glutamicibacter creatinolyticus, and Dietzia spp. in a horse reared in Italy.

Proteomic fingerprinting for the fast and accurate identification of species in the Polyporoid and Hymenochaetoid fungi clades.

Basidiomycotan fungi play significant roles in the biogeochemical cycle of carbon as wood decomposers and are used in the food industry for mushroom production and in biotechnology for the production of diverse bioactive compounds and bioremediation. The correct identification of basidiomycotan isolates is crucial for understanding their biology and being able to expand their applications. Currently, the identification of these organisms is performed by analyzing morphological and genomic characteristics, primarily those based on DNA biomarkers. Despite their efficiency, such methods require considerable expertise and are both time-consuming and error-prone (multistep workflow). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has emerged in the last decade as an accurate, fast, and powerful alternative for the identification of microorganisms. MALDI-TOF MS has been widely applied for the identification and taxonomical characterization of both bacteria and ascomycotan fungi from clinical origins. However, species of Basidiomycota have been poorly evaluated using this method. In the present study, we assessed the performance of MALDI-TOF MS using basidiomycotan isolates of two distinct taxonomical families: Polyporaceae and Hymenochaetaceae. Using a simple protocol, which eliminates the protein extraction step, we obtained high-quality mass spectra data and demonstrated that this method is efficient for the discrimination of isolates at the species level. SIGNIFICANCE: In this study, the MALDI-TOF mass spectrometry was employed to test its accuracy on the recognition of fungal species with high biotechnological and environmental interest. Using a simple and fast protocol, we obtained high-quality mass-spectra (protein fingerprinting) and proved that MALDI-TOF MS is sufficiently robust to the identification at species level and to evaluate the relationships among the isolates of the polyporoid and hymenochaetoid clades (Basidiomycota).

Severe outbreak of Centrocestus formosanus (Trematoda: Heterophyidae) in farm-raised ornamental platies Xiphophorus maculatus.

This report describes a severe outbreak of the gill fluke Centrocestus formosanus in farm-raised platies Xiphophorus maculatus in Brazil, with mortality rate approaching 95%. Typical clinical signs of infection were observed, with microscopic examinations of fresh gills revealing multiple cysts containing a once-folded metacercaria with an X-shaped excretory bladder. The 18S subunit of the metacercariae (BR1) was amplified by PCR, sequenced and analyzed by BLAST. Subsequent phylogenetic analyses revealed that the BR1 isolate was closely related to C. formosanus from Thailand. This is the first report of C. formosanus in ornamental fish in Brazil. Our observations suggest that platies are highly sensitive to this digenetic parasite. Controlling population densities of the parasite's intermediate host, the snail Melanoides tuberculata, would help to reduce outbreaks.

Gut microbiome modulation during treatment of mucositis with the dairy bacterium Lactococcus lactis and recombinant strain secreting human antimicrobial PAP.

Mucositis is an inflammatory condition of the gut, caused by an adverse effect of chemotherapy drugs, such as 5-fluorouracil (5-FU). In an attempt to develop alternative treatments for the disease, several research groups have proposed the use of probiotics, in particular, Lactic Acid Bacteria (LAB). In this context, the use of recombinant LAB, for delivering anti-inflammatory compounds has also been explored. In previous work, we demonstrated that either Lactococcus lactis NZ9000 or a recombinant strain expressing an antimicrobial peptide involved in human gut homeostasis, the Pancreatitis-associated Protein (PAP), could ameliorate 5-FU-induced mucositis in mice. However, the impact of these strains on the gut microbiota still needs to be elucidated. Therefore, in the present study, we aimed to characterize the effects of both Lactococci strains in the gut microbiome of mice through a 16 S rRNA gene sequencing metagenomic approach. Our data show 5-FU caused a significant decrease in protective bacteria and increase of several bacteria associated with pro-inflammatory traits. The Lactococci strains were shown to reduce several potential opportunistic microbes, while PAP delivery was able to suppress the growth of Enterobacteriaceae during inflammation. We conclude the strain secreting antimicrobial PAP was more effective in the control of 5-FU-dysbiosis.

The Druggable Pocketome of Corynebacterium diphtheriae: A New Approach for in silico Putative Druggable Targets.

Diphtheria is an acute and highly infectious disease, previously regarded as endemic in nature but vaccine-preventable, is caused by Corynebacterium diphtheriae (Cd). In this work, we used an in silico approach along the 13 complete genome sequences of C. diphtheriae followed by a computational assessment of structural information of the binding sites to characterize the "pocketome druggability." To this end, we first computed the "modelome" (3D structures of a complete genome) of a randomly selected reference strain Cd NCTC13129; that had 13,763 open reading frames (ORFs) and resulted in 1,253 (∼9%) structure models. The amino acid sequences of these modeled structures were compared with the remaining 12 genomes and consequently, 438 conserved protein sequences were obtained. The RCSB-PDB database was consulted to check the template structures for these conserved proteins and as a result, 401 adequate 3D models were obtained. We subsequently predicted the protein pockets for the obtained set of models and kept only the conserved pockets that had highly druggable (HD) values (137 across all strains). Later, an off-target host homology analyses was performed considering the human proteome using NCBI database. Furthermore, the gene essentiality analysis was carried out that gave a final set of 10-conserved targets possessing highly druggable protein pockets. To check the target identification robustness of the pipeline used in this work, we crosschecked the final target list with another in-house target identification approach for C. diphtheriae thereby obtaining three common targets, these were; hisE-phosphoribosyl-ATP pyrophosphatase, glpX-fructose 1,6-bisphosphatase II, and rpsH-30S ribosomal protein S8. Our predicted results suggest that the in silico approach used could potentially aid in experimental polypharmacological target determination in C. diphtheriae and other pathogens, thereby, might complement the existing and new drug-discovery pipelines.

Secretion of biologically active pancreatitis-associated protein I (PAP) by genetically modified dairy Lactococcus lactis NZ9000 in the prevention of intestinal mucositis.

BACKGROUND: Mucositis is one of the most relevant gastrointestinal inflammatory conditions in humans, generated by the use of chemotherapy drugs, such as 5-fluoracil (5-FU). 5-FU-induced mucositis affects 80% of patients undergoing oncological treatment causing mucosal gut dysfunctions and great discomfort. As current therapy drugs presents limitations in alleviating mucositis symptoms, alternative strategies are being pursued. Recent studies have shown that the antimicrobial pancreatitis-associated protein (PAP) has a protective role in intestinal inflammatory processes. Indeed, it was demonstrated that a recombinant strain of Lactococcus lactis expressing human PAP (LL-PAP) could prevent and improve murine DNBS-induced colitis, an inflammatory bowel disease (IBD) that causes severe inflammation of the colon. Hence, in this study we sought to evaluate the protective effects of LL-PAP on 5-FU-induced experimental mucositis in BALB/c mice as a novel approach to treat the disease. RESULTS: Our results show that non-recombinant L. lactis NZ9000 have antagonistic activity, in vitro, against the enteroinvasive gastrointestinal pathogen L. monocytogenes and confirmed PAP inhibitory effect against Opportunistic E. faecalis. Moreover, L. lactis was able to prevent histological damage, reduce neutrophil and eosinophil infiltration and secretory Immunoglobulin-A in mice injected with 5-FU. Recombinant lactococci carrying antimicrobial PAP did not improve those markers of inflammation, although its expression was associated with villous architecture preservation and increased secretory granules density inside Paneth cells in response to 5-FU inflammation. CONCLUSIONS: We have demonstrated for the first time that L. lactis NZ9000 by itself, is able to prevent 5-FU-induced intestinal inflammation in BALB/c mice. Moreover, PAP delivered by recombinant L. lactis strain showed additional protective effects in mice epithelium, revealing to be a promising strategy to treat intestinal mucositis.

Complete genome sequences of Francisella noatunensis subsp. orientalis strains FNO12, FNO24 and FNO190: a fish pathogen with genomic clonal behavior.

The genus Francisella is composed of Gram-negative, pleomorphic, strictly aerobic and non-motile bacteria, which are capable of infecting a variety of terrestrial and aquatic animals, among which Francisella noatunensis subsp. orientalis stands out as the causative agent of pyogranulomatous and granulomatous infections in fish. Accordingly, F. noatunensis subsp. orientalis is responsible for high mortality rates in freshwater fish, especially Nile Tilapia. In the current study, we present the genome sequences of F. noatunensis subsp. orientalis strains FNO12, FNO24 and FNO190. The genomes include one circular chromosome of 1,859,720 bp, consisting of 32 % GC content, 1538 coded proteins and 363 pseudogenes for FNO12; one circular chromosome of 1,862,322 bp, consisting of 32 % GC content, 1537 coded proteins and 365 pseudogenes for FNO24; and one circular chromosome of 1,859,595 bp, consisting of 32 % GC content, 1539 coded proteins and 362 pseudogenes for FNO190. All genomes have similar genetic content, implicating a clonal-like behavior for this species.