Effect of allogeneic pure platelet-rich plasma, sodium cloxacillin, and their combination for the treatment of subclinical mastitis in crossbred cows.

Introduction: Bovine subclinical mastitis (SCM) caused by Gram-positive bacteria is a major cause of economic loss in the dairy industry, exacerbated in situations where antimicrobial resistance is present. Pure platelet-rich plasma (P-PRP) may be a therapeutic alternative for SCM, when used alone or with antibiotics, such as sodium cloxacillin (SC). This study aimed 1) to evaluate the therapeutic efficacy of allogeneic P-PRP, SC, and their combination (P-PRP+SC) in cows with SCM caused by Staphylococcus aureus and by streptococci (Staphylococcus aureus and S. dysgalactiae); 2) to determine the concentrations of somatic cells (SCC), interleukin 1 beta (IL-1ß), tumor necrosis factor-alpha (TNF-α) and TGF-ß1 in milk samples of the cows. Methods: 130 cows from 4 dairy herds completed the study, of which 40 cows were treated with P-PRP (10 mL), 28 cows with SC (5g), 36 with P-PRP+SC (10mL/5g), and 26 did not receive no treatment (negative control group, NCG). Results: The overall bacteriological cure was observed in 10/40 (25%) cows in the P-PRP group, 9/28 (32.14%) animals in the SC group, 26/36 (72.22%) cows in the P-PRP+SC group, and 10/26 (38.46%) animals in the NCG. SCM caused by S. aureus (82/130, 63.08%), was cured in 6/24 (25%) cows treated with P-PRP, 7/24 (29.2%) cows treated with SC, 8/16 (50%) animals treated with P-PRP+SC, and in 8/18 (44.4%) cows in NCG. For SCM caused by the streptococci (48/130, 36.91%), the cure was achieved in 4/12 (33.3%) cows treated with P-PRP, 2/4 (50%) cows treated with SC, 18/20 (90%) cows treated with P-PRP+SC, and in 2/8 (25%) cows of the NCG. SCC was significantly (p < 0.001) affected by the treatment, herd, cure, bacteria group, and number of calvings factors. IL-1ß milk concentrations were significantly (p < 0.001) influenced by treatment and farm factors, and the interaction between these factors. TNF-α milk concentrations were significantly (p < 0.001) influenced by time factor. TGF-ß1 milk concentrations were significantly affected by the time and cure factors. Conclusion: The combination of P-PRP and SC showed the best therapeutic response (90%) against bovine SCM caused by streptococci. However, none of the treatments showed an effective therapeutic response against S. aureus.

A systematic review of diabetic foot infections: pathogenesis, diagnosis, and management strategies.

Background: Diabetic foot infection represents a significant complication of diabetes mellitus, contributing substantially to morbidity, mortality, and healthcare expenditure worldwide. Accurate diagnosis relies on a comprehensive assessment integrating clinical evaluation, imaging studies, and microbiological analysis. Management necessitates a multidisciplinary approach, encompassing surgical intervention, antimicrobial therapy, and advanced wound care strategies. Preventive measures are paramount in reducing the incidence and severity, emphasizing patient education, regular foot screenings, and early intervention. Methods: The researchers performed a systematic review of literature using PUBMED MESH keywords. Additionally, the study was registered in the International Prospective Register of Systematic Reviews at the Center for Reviews and Dissemination, University of York (CRD42021277788). This review provides a comprehensive overview of the microbial spectrum and antibiotic susceptibility patterns observed in diabetic foot infections. Results: The search through the databases finally identified 13 articles with 2545 patients from 2021 to 2023. Overall, the predominant Gram-positive microbial species isolated were Staphylococcus aureus, Enterococcus fecalis, Streptococcus pyogenes, Streptococcus agalactiae, and Staphylococcus epidermidis. Whereas the predominant Gram-negative included Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis and Pseudomonas aeruginosa. Conclusion: Diabetic foot infections represent a complex and multifaceted clinical entity, necessitating a holistic approach to diagnosis, management, and prevention. Limited high-quality research data on outcomes and the effectiveness of guideline recommendations pose challenges in updating and refining existing DFI management guidelines. Systematic review registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42021277788, identifier CRD42021277788.

Mimicry of Rhabdomyosarcoma by Tonsillar Actinomycosis: Case Report.

Actinomycosis is a rare infectious disease characterized by slowly progressive, chronic suppurative lesions, often mistaken for malignancies due to its ability to mimic them. It is caused by Actinomyces bacteria, which are part of the normal flora of the human oropharynx, gastrointestinal, and urogenital tracts. This case report describes a 51-year-old male with a history of mandibular rhabdomyosarcoma presenting with severe shoulder and hip pain, dysphagia, and headaches, initially suspected to be a cancer recurrence. However, after further investigation, including a PET-CT and tonsillectomy, the diagnosis of actinomycosis was confirmed through histopathological examination. The case highlights the diagnostic challenges of actinomycosis, especially in patients with complex clinical histories, emphasizing the importance of considering it as a differential diagnosis in similar presentations. The patient was treated with long-term antibiotic therapy, predominantly beta-lactams, demonstrating the necessity of a comprehensive diagnostic approach and the implications of a delayed diagnosis. This case underscores the critical need for high clinical suspicion and awareness among healthcare professionals regarding the potential for actinomycosis to mimic more common diseases, ensuring timely and accurate treatment.

Listeria monocytogenes in beef: a hidden risk.

Listeria monocytogenes in beef receives less attention compared to other pathogens such as Salmonella and Escherichia coli. To address this gap, we conducted a literature review focusing on the presence of L. monocytogenes in beef. This review encompasses the pathogenic mechanisms, routes of contamination, prevalence rates, and the laws and regulations employed in various countries. Our findings reveal a prevalence of L. monocytogenes in beef and beef products ranging from 2.5% to 59.4%. Notably, serotype 4b was most frequently isolated in cases of beef contamination during food processing, with the skinning and evisceration stages identified as critical points of contamination.

Diversity and Bioprospection of Gram-positive Bacteria Derived from a Mayan Sinkhole.

Water-filled sinkholes known locally as cenotes, found on the Yucatán Peninsula, have remarkable biodiversity. The primary objective of this study was to explore the biotechnological potential of Gram-positive cultivable bacteria obtained from sediment samples collected at the coastal cenote Pol-Ac in Yucatán, Mexico. Specifically, the investigation aimed to assess production of hydrolytic enzymes and antimicrobial compounds. 16 S rRNA gene sequencing led to the identification of 49 Gram-positive bacterial isolates belonging to the phyla Bacillota (n = 29) and Actinomycetota (n = 20) divided into the common genera Bacillus and Streptomyces, as well as the genera Virgibacillus, Halobacillus, Metabacillus, Solibacillus, Neobacillus, Rossellomorea, Nocardiopsis and Corynebacterium. With growth at 55ºC, 21 of the 49 strains were classified as moderately thermotolerant. All strains were classified as halotolerant and 24 were dependent on marine water for growth. Screening for six extracellular hydrolytic enzymes revealed gelatinase, amylase, lipase, cellulase, protease and chitinase activities in 93.9%, 67.3%, 63.3%, 59.2%, 59.2% and 38.8%, of isolated strains, respectively. The genes for polyketide synthases type I, were detected in 24 of the strains. Of 18 strains that achieved > 25% inhibition of growth in the bacterial pathogen Staphylococcus aureus ATCC 6538, 4 also inhibited growth in Escherichia coli ATCC 35,218. Isolates Streptomyces sp. NCA_378 and Bacillus sp. NCA_374 demonstrated 50-75% growth inhibition against at least one of the two pathogens tested, along with significant enzymatic activity across all six extracellular enzymes. This is the first comprehensive report on the biotechnological potential of Gram-positive bacteria isolated from sediments in the cenotes of the Yucatán Peninsula.

Effects of antimicrobial photodynamic therapy with photodithazine® on methicillin-resistant Staphylococcus aureus (MRSA): Studies in biofilms and experimental model with Galleria mellonella.

Staphylococcus aureus infections are a severe health problem due to the high mortality rate. Conventional treatment of these infections is via the administration of antibiotics. However, its indiscriminate use can select resistant microorganisms. Thus, it is necessary to develop alternatives for antibiotic therapy. Antimicrobial Photodynamic Therapy (aPDT), a therapeutic method that associates a photosensitizer (PS), a light source with adequate wavelength to the PS, interacts with molecular oxygen generating reactive oxygen species responsible for cell inactivation, is a viable alternative. This work aimed to analyze, in vitro and in vivo, the action of aPDT with PS Photodithazine® (PDZ) on the methicillin-resistant S. aureus (MRSA) strain. In the in vitro method, the S. aureus biofilm was incubated with PDZ at 50 and 75 µg.mL-1 for 15 min, adopting the light dose of 25, 50, and 100 J/cm2. In addition, PS interaction, formation of reactive oxygen species (ROS), bacterial metabolism, adhesion, bacterial viability, and biofilm structure were evaluated by scanning electron microscopy. Subsequently, the strain was inoculated into models of Galleria mellonella, and the survival curve, health scale, blood cell analysis, and CFU/mL of S. aureus in the hemolymph were analyzed after aPDT. In the in vitro results, bacterial reduction was observed in the different PDZ concentrations, highlighting the parameters of 75 µg.mL-1 of PDZ and 100 J/cm2. As for in vivo results, aPDT increased survival and stimulated the immune system of G. mellonella infected by S. aureus. aPDT proved effective in both models, demonstrating its potential as an alternative therapy in treating MRSA bacterial infections.

Prevalence of Gram positive bacteria in the affected individuals of Otitis media with effusion from the indigenous population of Southern Punjab, Pakistan: first report / Prevalência de bactérias Gram-positivas em indivíduos afetados por Otite média com efusão da população indígena do sul de Punjab, Paquistão: primeiro relatório

Otitis media with effusion (OME) is a type of otitis media (OM) characterized by the presence of fluid behind intact tympanic membrane and is one of the most common diseases of early childhood. It is an infectious disease associated with the presence of many pathogenic bacteria in the middle ear of affected individuals. This study was aimed to determine the prevalence of Gram-positive bacteria from the middle ear of OME patients in the population of Southern Punjab, Pakistan. The incidence of OME under comprehensive healthcare setting was investigated in patients who consulted at the department of ear, throat and nose, Bahawal Victoria Hospital (BVH), Bahawalpur, from December, 2019 to May, 2021. Ear swabs were taken from affected and normal individuals. After culturing bacteria from the ear swabs, microscopic analysis and biochemical tests were performed to characterize the cultured Gram-positive bacteria. Out of 352 patients examined, 109 (30.9%) patients had OME. Age of the participants ranged from 14 to 50 years; individuals between the ages of 14 and 22 years had the highest infection rates, while individuals between 40 and 50 years had the lowest rate of infection. Tympanic membrane perforation, fever, cough, sore throat, ear pain and hearing problem showed association with symptoms of OME. Microscopic analysis and biochemical characterization showed the presence of streptococci and staphylococci in all the studied samples. The most frequently isolated bacteria were Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus with percentage of 53.3%, 20% and 13.3% respectively. Enterococcus faecalis (6.6%) and Staphylococcus epidermidis (6.6%) were also identified in the studied samples. This study will help in the better medical administration of OME affected individuals.

A otite média com efusão (OME) é um tipo de otite média (OM) caracterizada pela presença de líquido atrás da membrana timpânica intacta e é uma das doenças mais comuns durante a primeira infância. Trata-se de uma doença infecciosa associada à presença de muitas bactérias patogênicas na orelha média dos indivíduos afetados. Este estudo teve como objetivo determinar a prevalência de bactérias Gram-positivas do ouvido médio de pacientes com OME na população do sul de Punjab, Paquistão. A incidência de OME em ambiente de saúde abrangente foi investigada em pacientes que consultaram no departamento de ouvido, garganta e nariz, Hospital Bahawal Victoria (BVH), Bahawalpur, de dezembro de 2019 a maio de 2021. Cotonetes de orelha foram coletados de indivíduos afetados e normais. Após a cultura das bactérias dos swabs auriculares, análises microscópicas e testes bioquímicos foram realizados para caracterizar as bactérias Gram-positivas cultivadas. Dos 352 pacientes examinados, 109 (30,9%) apresentavam OME. A idade dos participantes variou de 14 a 50 anos; indivíduos entre 14 e 22 anos apresentaram as maiores taxas de infecção, enquanto indivíduos entre 40 e 50 anos apresentaram as menores taxas de infecção. Perfuração da membrana timpânica, febre, tosse, dor de garganta, dor de ouvido e problema de audição apresentaram associação com sintomas de OME. A análise microscópica e a caracterização bioquímica mostraram a presença de estreptococos e estafilococos em todas as amostras estudadas. As bactérias isoladas com maior frequência foram Streptococcus pneumoniae, Streptococcus pyogenes e Staphylococcus aureus com percentuais de 53,3%, 20% e 13,3%, respectivamente. Enterococcus faecalis (6,6%) e Staphylococcus epidermidis (6,6%) também foram identificados nas amostras estudadas. Este estudo ajudará na melhor administração médica de indivíduos afetados por OME.

Antibacterial effect of Asphodelus fistulosus aqueous and ethanolic crude extracts on gram positive and gram negative bacteria / Efeito antibacteriano de extratos brutos aquosos e etanólicos de Asphodelus fistulosus sobre bactérias gram-positivas e gram-negativas

Asphodelus fistulosus (A. fistulosus) is a wild plant grows in Jordan. Traditionally, it is used to treat different medical conditions and diseases such as respiratory ailments, against burns and dermatomucosal infections.This study aims to find out the effects of A. fistulosus aqueous and ethanolic crude extracts on Staphylococcus aureus(S. aureus) as gram positive bacteria and Escherichia coli (E. coli) as gram negative bacteria and to investigate which one will be affected either by aqueous and/or ethanolic crude extracts of A. fistulosus shooting parts that were collected from Jerash in the north of Jordan. Agar well diffusion method was used to evaluate the antibacterial activity of the crude extracts. In addition, MIC (minimum inhibitory concentration) as well as MBC (minimum bactericidal concentration) were determined against both types of bacteria. The results showed that flower aqueous extract of A. fistulosus was very effective against E. coli (20.0 ± 0.50) mm and caused a (14.0 ± 0.50) mm inhibition to S. aureus. The ethanolic extract of stem was very effective cauesed a (19.0 ± 0.50) mm inhibition in both bacterial species. Respectively, both S. aureus and E. coli were inhibited by ethanolic and aqueous extracts (mixture1 and mixture2) (15.0 ± 0.00 mm and 10.5 ± 0.50 mm). The highest antimbacterial activity was observed for the leaves aqueous extract against E.coli (0.06120 mg/mL). The obtained MIC values from A. fistulosus parts extracts demonstrated antibacterial activity ranged between 7.606 and 0.06120 mg/mL. The highest antimicrobial activity was recorded in the leaves aqueous extract against E. coli.The MBC value of stem aqueous extract was 5.00 mg/mL against both S. aureus and E. coli. On the other hand, ethanolic and aqueous extracts of the leaves gave MBC values 5.00 mg/mL, and 0.156 mg/mL, respectively, against E. coli.Based on the results of this study, it can be concluded that there is good inhibitory effect of aqueous and ethanolic of A. fistulosus shooting parts extracts on growth of E. coli and S. aureus. Adding to that, stem ethanolic extract has the most effective against S. aureus while aqueous extract of flower has the most effective against E. coli.So, it is recommended to have further future studies on the A. fistulosus shooting parts crude extract bioactive components and the mechanism of how these constituents affect these types of bacteria.

Asphodelus fistulosus (A. fistulosus) é uma planta selvagem que cresce na Jordânia. Tradicionalmente, é usada para tratar diferentes condições médicas e doenças, como doenças respiratórias, contra queimaduras e infecções dermatomucosas. Bactérias positivas e Escherichia coli (E. coli) como bactérias gram-negativas e investigar qual delas será afetada por extratos brutos aquosos e/ou etanólicos de partes de tiro de A. fistulosus que foram coletadas em Jerash no norte da Jordânia. O método de difusão em poço de ágar foi utilizado para avaliar a atividade antibacteriana dos extratos brutos. Além disso, MIC (concentração inibitória mínima) e MBC (concentração bactericida mínima) foram determinados contra ambos os tipos de bactérias. Os resultados mostraram que o extrato aquoso de flores de A. fistulosus foi muito eficaz contra E. coli (20,0 + 0,50 mm) e causou uma inibição (14,0 + 0,50 mm) para S. aureus. O extrato etanólico do caule foi muito eficaz, causando inibição (19,0 + 0,50 mm) em ambas as espécies bacterianas. Respectivamente, tanto S. aureus quanto E. coli foram inibidos pelos extratos etanólico e aquoso (mistura 1 e mistura 2) (15,0 + 0,00 mm e 10,5 + 0,50 mm). A maior atividade antibacteriana foi observada para o extrato aquoso das folhas contra E. coli (0,06120 mg/mL). Os valores de CIM obtidos dos extratos de partes de A. fistulosus demonstraram atividade antibacteriana variando entre 7,606 e 0,06120 mg/mL. A maior atividade antimicrobiana foi registrada no extrato aquoso das folhas contra E. coli. O valor de CBM do extrato aquoso do caule foi de 5,00 mg/mL contra S. aureus e E. coli. Por outro lado, os extratos etanólico e aquoso das folhas apresentaram valores de CBM de 5,00 mg/mL e 0,156 mg/mL, respectivamente, contra E. coli, efeito de extratos aquosos e etanólicos de partes de tiro de A. fistulosus no crescimento de E. coli e S. aureus. Somando-se a isso, o extrato etanólico do caule é o mais eficaz contra S. aureus enquanto o extrato aquoso da flor é o mais eficaz contra E. coli. Assim, recomenda-se a realização de estudos futuros sobre os componentes bioativos do extrato bruto de partes fotografais de A. fistulosus e o mecanismo de como esses constituintes afetam esses tipos de bactérias.

Multidrug Efflux System-mediated resistance in Staphylococcus aureus under a One Health approach.

The aim of the study was to track the spread of antimicrobial resistance among the different sectors of One Health through the detection of Multidrug-Efflux-System in multidrug-resistant Staphylococcus aureus isolates. Multidrug-resistant (MDR) and methicillin-resistant (MRSA) S. aureus isolates were selected: 25 of human, one of animal and eight of food origin. The efflux system genes norA, norB, norC, LmrS, tet38 and msrA were screened by PCR. The activity of the efflux systems was determined by the minimum inhibitory concentration (MIC) of tetracycline and ciprofloxacin in the presence and absence of CCCP and in the quantification of ethidium bromide efflux. Furthermore, biofilm formation was determined in the presence and absence of the CCCP. The molecular epidemiology of the isolates was traced with the aid of PFGE. The gene norC was the most prevalent, detected in all isolates and msrA was the least prevalent, detected in only two isolates from humans. There was no difference in the MICs of tetracycline and ciprofloxacin in the presence of CCCP, but 55.9% of isolates showed ethidium bromide efflux. The presence of CCCP decreased the biofilm formation. Regarding the molecular epidemiology, in three clusters was a mixture of the isolates from different origins. Therefore, S. aureus MDR with active multidrug efflux systems are circulating between One Health domains and it is necessary to consider strategies to decrease this circulation in order to prevent the dissemination of resistance mediated by MES.

Aquiluscidin, a Cathelicidin from Crotalus aquilus, and the Vcn-23 Derivative Peptide, Have Anti-Microbial Activity against Gram-Negative and Gram-Positive Bacteria.

Anti-microbial peptides play a vital role in the defense mechanisms of various organisms performing functions that range from the elimination of microorganisms, through diverse mechanisms, to the modulation of the immune response, providing protection to the host. Among these peptides, cathelicidins, a well-studied family of anti-microbial peptides, are found in various animal species, including reptiles. Due to the rise in anti-microbial resistance, these compounds have been suggested as potential candidates for developing new drugs. In this study, we identified and characterized a cathelicidin-like peptide called Aquiluscidin (Aq-CATH) from transcripts obtained from the skin and oral mucosa of the Querétaro's dark rattlesnake, Crotalus aquilus. The cDNA was cloned, sequenced, and yielded a 566-base-pair sequence. Using bioinformatics, we predicted that the peptide precursor contains a signal peptide, a 101-amino-acid conserved cathelin domain, an anionic region, and a 34-amino-acid mature peptide in the C-terminal region. Aq-CATH and a derived 23-amino-acid peptide (Vcn-23) were synthesized, and their anti-microbial activity was evaluated against various species of bacteria in in vitro assays. The minimal inhibitory concentrations against bacteria ranged from 2 to 8 µg/mL for both peptides. Furthermore, at concentrations of up to 50 µM, they exhibited no significant hemolytic activity (<2.3% and <1.2% for Aquiluscidin and Vcn-23, respectively) against rat erythrocytes and displayed no significant cytotoxic activity at low concentrations (>65% cell viability at 25 µM). Finally, this study represents the first identification of an antimicrobial peptide in Crotalus aquilus, which belongs to the cathelicidin family and exhibits the characteristic features of these peptides. Both Aq-CATH and its derived molecule, Vcn-23, displayed remarkable inhibitory activity against all tested bacteria, highlighting their potential as promising candidates for further antimicrobial research.

Enzymatic synthesis of new antimicrobial peptides for food purposes.

Growing consumer awareness of the potential negative health effects of synthetic antibiotics has prompted the search for more natural preservatives that can improve the safety and quality of food. In this study we report the enzymatic synthesis of N-α-[Carbobenzyloxy]-Ile-Gln (Z-IQ) which is the precursor of Ile-Gln (IQ), a new antibacterial dipeptide, using an aqueous-organic biphasic system formed by 50% (v/v) ethyl acetate in 0.1 M Tris - HCl buffer pH 8. A partially purified proteolytic extract from the fruits of Solanum granuloso leprosum, named granulosain, proved to be a robust biocatalyst for the synthesis of Z-IQ, eliciting 71 ± 0.10% maximal peptide yield in the above described conditions. After cleaving and purifying IQ dipeptide, antimicrobial activity was assayed against Staphylococcus aureus ATCC 25923, Staphylococcus hominis A17771, and Staphylococcus aureus C00195, and MIC values between 118 ± 0.01 µg/mL and 133.7 ± 0.05 µg/mL were obtained. In addition, IQ showed MIC of 82.4 ± 0.01 µg/mL and 85.0 ± 0.00 µg/mL against Escherichia coli ATCC 25922 and Escherichia coli A17683, respectively. IQ did not show inhibitory activity against single-drug resistance (SDR) strains, such as Klebsiella oxytoca A19438 (SDR) and Pseudomonas aeruginosa C00213 (SDR), and against multidrug-resistant Enterococcus faecalis I00125 (MDR). IQ also caused growth inhibition of Helicobacter pylori NCTC 11638 and three wild-type H. pylori strains, which are sensitive to AML, MTZ, LEV and CLA (H. pylori 659), resistant to LEV (H. pylori 661 SDR), and resistant to MTZ (H. pylori 662 SDR). Finally, this study contributes with a new dipeptide (IQ) that can be used as an antimicrobial agent for food preservation or as a safe ingredient of functional foods.

Detection of Cutibacterium acnes in Tissue Samples from Clean Primary Shoulder Surgeries - Part II.

Objective Research and identification of Cutibacterium acnes ( C. acnes ) and other microorganisms in deep tissue samples collected in clean shoulder surgeries of patients who did not undergo any previous invasive joint procedure and who had no clinical history of infection. Methods We analyzed the results of cultures of intraoperative deep tissue samples from 84 patients submitted to primary clean shoulder surgery. Tubes containing culture medium were used for storage and transport of anaerobic agents, prolonged incubation time, and mass spectrometer for diagnosis of bacterial agents. Results Bacteria growth was evidenced in 34 patients (40.4%) of the 84 included in the study. Of these, 23 had growth of C. acnes in at least one sample of deep tissue collected, corresponding to 27.3% of the total patients. The second most common agent was Staphylococcus epidermidis , present in 7.2% of the total individuals included. We showed a higher relationship between sample positivity and males, a lower mean age, absence of diabetes mellitus, ASA I score, and antibiotic prophylaxis in anesthetic induction with cefuroxime. Conclusions A high percentage of isolates of different bacteria was found in shoulder tissue samples of patients undergoing clean and primary surgeries, who had no history of previous infection. Identification of C. acnes was high (27.6%), and Staphylococcus epidermidis was the second most frequent agent (7.2%).

Bactericidal activity of silver nanoparticles in drug-resistant bacteria.

Bacterial resistance to multiple drugs is a worldwide problem that afflicts public health. Various studies have shown that silver nanoparticles are good bactericidal agents against bacteria due to the adherence and penetration of the external bacterial membrane, preventing different vital functions and subsequently bacterial cell death. A systematic review of ScienceDirect, PubMed, and EBSCOhost was conducted to synthesize the literature evidence on the association between the bactericidal property of silver nanoparticles on both resistant Gram-positive and Gram-negative bacteria. Eligible studies were original, comparative observational studies that reported results on drug-resistant bacteria. Two independent reviewers extracted the relevant information. Out of the initial 1 420, 142 studies met the inclusion criteria and were included to form the basis of the analysis. Full-text screening led to the selection of 6 articles for review. The results of this systematic review showed that silver nanoparticles act primarily as bacteriostatic agents and subsequently as bactericides, both in Gram-positive and Gram-negative drug-resistant bacteria.

Antimicrobial photodynamic in vitro inactivation of Enterococcus spp. and Staphylococcus spp. strains using tetra-cationic platinum(II) porphyrins.

This manuscript presents the first report on antimicrobial photo-inactivation in vitro using tetra-cationic porphyrins with peripheral platinum(II) bipyridyl complexes against Gram-positive bacteria. Two isomeric tetra-cationic porphyrins (3TPyP and 4TPyP) were tested against clinically important bacterial species. The antimicrobial activity assays were performed at specific photosensitizer (PS) concentrations under dark and white-light LED irradiation conditions for 120 min. The porphyrin 3-PtTPyP was the most efficient PS against the bacteria tested, inhibiting bacterial growth in just 15 min and 30 min at low concentrations (3.75 and 0.45 µM). The minimal inhibitory concentration of the porphyrin increased in the presence of reactive oxygen species scavengers, indicating that singlet oxygen and radical species likely participated in the photo-oxidation mechanism. In addition, the checkerboard assay that tests the association of compounds, showed a synergistic effect, suggesting a potentiation of the antibacterial effect when porphyrin was tested in combination with ciprofloxacin and vancomycin. Thus, tetra-cationic porphyrins containing platinum(II) complexes are promising agents for microbial photo-inactivation as an alternative therapy against infections.

Detection of Cutibacterium acnes in Tissue Samples from Clean Primary Shoulder Surgeries - Part II / Detecção de Cutibacterium acnes em amostras de tecidos de cirurgias limpas primárias do ombro - Parte II

Abstract Objective Research and identification of Cutibacterium acnes (C. acnes) and other microorganisms in deeptissue samples collected in clean shoulder surgeries of patients who did not undergo any previous invasive joint procedure and who had no clinical history of infection. Methods We analyzed the results of cultures of intraoperative deep tissue samples from 84 patients submitted to primary clean shoulder surgery. Tubes containing culture medium were used for storage and transport of anaerobic agents, prolonged incubation time, and mass spectrometer for diagnosis of bacterial agents. Results Bacteria growth was evidenced in 34 patients (40.4%) of the 84 included in the study. Of these, 23 had growth of C. acnes in at least one sample of deep tissue collected, corresponding to 27.3% of the total patients. The second most common agent was Staphylococcus epidermidis, present in 7.2% of the total individuals included. We showed a higher relationship between sample positivity and males, a lower mean age, absence of diabetes mellitus, ASA I score, and antibiotic prophylaxis in anesthetic induction with cefuroxime. Conclusions A high percentage of isolates of different bacteria was found in shoulder tissue samples of patients undergoing clean and primary surgeries, who had no history of previous infection. Identification of C. acnes was high (27.6%), and Staphylococcus epidermidis was the second most frequent agent (7.2%).

Resumo Objetivo Pesquisa e identificação de Cutibacterium acnes (C. acnes) e de outros microrganismos em amostras de tecidos profundos coletados em cirurgias limpas de ombro em pacientes que não foram submetidos a nenhum procedimento invasivo articular prévio e que não possuíam antecedentes clínicos de infecção. Métodos Foram analisados os resultados das culturas de amostras de tecidos profundos intraoperatórias de 84 pacientes submetidos à cirurgia limpa primária do ombro. Foram utilizados tubos contendo meio de cultivo para armazenamento e transporte de agentes anaeróbicos, tempo prolongado de incubação e espectrômetro de massa para diagnósticos de agentes bacterianos. Resultados Foi evidenciado o crescimento de bactérias em 34 pacientes (40,4%) dos 84 incluídos no estudo. Desses, 23 apresentavam crescimento de C. acnes em pelo menos uma amostra de tecido profundo coletada, correspondendo a 27,3% do total de pacientes. O segundo agente mais encontrado foi o Staphylococcus epidermidis, presente em 7,2% do total de indivíduos incluídos. Evidenciamos maior relação da positividade de amostras com o gênero masculino, uma média de idade inferior, a ausência de diabetes mellitus, o escore ASA I e a profilaxia antibiótica na indução anestésica com cefuroxima. Conclusões Verificou-se um elevado percentual de isolados de diferentes bactérias em amostras de tecidos de ombros de pacientes submetidos a cirurgias limpas e primárias e sem histórico de infecção anterior. A identificação de C. acnes foi elevada (27,6%) e o Staphylococcus epidermidis foi o segundo agente mais frequente (7,2%).

Empiema pleural por Parvimonas micra en un paciente inmunocompetente: reporte de un caso / Pleural empyema by Parvimonas micra in an immunocompetent patient: a case report

Se presenta el caso de un paciente joven inmunocompetente, con antecedente de tuberculosis pulmonar, que acude al hospital por un cuadro clínico subagudo de fiebre persistente, baja de peso, disnea y abolición del murmullo vesicular. La tomografía de tórax mostró un extenso empiema en hemitórax izquierdo. Se le toman muestras para detección de gérmenes comunes y se le colocan un tubo de drenaje torácico y se inicia antibioticoterapia. La prueba de MALDI-TOF MS identificó a Parvimonas micra, una bacteria anaerobia, comensal de la flora oral, asociado a periodontitis severa, escasamente reportado en empiema pleural, especialmente, en personas inmunocompetentes. En la evaluación odontológica se realizó el diagnóstico de gingivitis y pericoronaritis de la tercera molar. El paciente evolucionó favorablemente. Se sugiere que, en casos de empiemas pleurales subagudos o crónicos, se debe considerar, además de las micobacterias, como agente etiológico al Parvimonas micra, y optar por exámenes como MALDI-TOF MS o secuenciamiento del 16S rRNA, colocación de tubo de tórax, cobertura antibiótica empírica y evaluación odontológica.

We present the case of a young immunocompetent patient, with a history of pulmonary tuberculosis, who attended the hospital with a subacute clinical picture of persistent fever, weight loss, dyspnea and abolition of vesicular murmur. Chest CT scan showed an extensive empyema in the left hemithorax. Samples were taken for detection of common germs. Then, a chest drainage tube was placed and antibiotic therapy started. The MALDI-TOF MS test identified Parvimonas micra, an anaerobic bacterium, commensal to the oral flora, associated with severe periodontitis, but rarely reported in cases of pleural empyema, especially in immunocompetent patients. Gingivitis and pericoronaritis of the third molar were diagnosed during oral evaluation. The patient progressed favorably. Parvimonas micra should be considered as a possible etiological agent in cases of subacute or chronic pleural empyema, in addition to mycobacteria. Tests such as MALDI-TOF MS or 16S rRNA sequencing, chest tube placement, empirical antibiotic coverage and an adequate oral evaluation should be considered in these cases.

Evaluación de la formación de biopelículas en aislamientos bacterianos y fúngicos por el método semicuantitativo de microtitulación con cristal violeta y el cualitativo de agar con rojo Congo / Assessment of biofilms formation of bacterial and fungal isolates using qualitative Congo red agar and semiquantitative crystal violet microtiter methods

Introducción. El 65 % de las infecciones humanas son producidas por bacterias o levaduras, cuya capacidad de formar biopelículas las hace más resistentes a los antimicrobianos y antifúngicos. Objetivo. Determinar la capacidad de formación de biopelículas en aislamientos bacterianos y fúngicos por medio de los métodos cuantitativo de microtitulación con cristal violeta y cualitativo de cultivo en agar con rojo Congo. Materiales y métodos. Con el método cuantitativo, se utilizaron los medios de cultivo infusión cerebro-corazón, tripticasa de soya y Müeller-Hinton para aislamientos bacterianos; para levaduras, se usaron caldo infusión cerebro-corazón y Sabouraud dextrosa. Para el método cualitativo de cultivo en agar, se utilizaron los mismos medios de cultivo más una solución con 3 % de rojo Congo y 10 % de dextrosa. Cómo método de referencia, se utilizó la propuesta de Stepanovic et al. Resultados. Se evaluaron 103 aislamientos bacterianos y 108 de levaduras. No es recomendable sustituir el caldo infusión cerebro-corazón por los caldos tripticasa de soya y Müeller-Hinton en el método cuantitativo, para evaluar la formación de biopelículas en los aislamientos bacterianos. El medio Sabouraud dextrosa, en caldo y agar, puede sustituir al de infusión de cerebro-corazón para evaluar la formación de biopelículas en levaduras, tanto por el método cuantitativo como por el cualitativo. Conclusión. El estudio de las biopelículas en el laboratorio de microbiología, a partir del método cualitativo de cultivo en agar con rojo Congo, es un procedimiento sencillo, rápido y de bajo costo, que proporciona información útil para el diagnóstico y la terapéutica de infecciones persistentes causadas por bacterias y levaduras.

Introduction. Sixty-five percent of human infections are caused by bacteria or yeasts able to form biofilms. This feature makes them more resistant to antimicrobials and antifungals. Objective. To determine biofilm formation capacity of bacterial and fungal isolates by quantitative crystal violet microtiter and qualitative Congo red agar methods. Materials and methods. Brain-heart infusion, trypticase soy broth and Müeller-Hinton culture media were used in bacterial isolates for the quantitative method; brain-heart infusion broth and Sabouraud dextrose were used for yeasts. The same culture media plus 3% Congo red and 10% dextrose were used to apply the qualitative method in agar. The proposal by Stepanovic, et al. was used as a reference method. Results. We evaluated 103 bacterial isolates and 108 yeasts isolates. We did not recommend substitute brain-heart infusion broth for trypticase soy and Müeller-Hinton broths for biofilm formation assessment in bacterial isolates using the quantitative method. Sabouraud dextrose medium, both broth and agar, can replace brain-heart infusion to assess biofilm formation in yeasts, quantitatively and qualitatively. Conclusion. The study of biofilms in the microbiology laboratory, using Congo red agar qualitative method, is a simple, fast, and inexpensive procedure that provides precise information for the diagnosis and treatment of persistent infections caused by bacteria and yeasts.

In Silico and In Vitro Identification of 1,8-Dihydroxy-4,5-dinitroanthraquinone as a New Antibacterial Agent against Staphylococcus aureus and Enterococcus faecalis.

Increasing rates of bacterial resistance to antibiotics are a growing concern worldwide. The search for potential new antibiotics has included several natural products such as anthraquinones. However, comparatively less attention has been given to anthraquinones that exhibit functional groups that are uncommon in nature. In this work, 114 anthraquinones were evaluated using in silico methods to identify inhibitors of the enzyme phosphopantetheine adenylyltransferase (PPAT) of Staphylococcus aureus, Enterococcus faecalis, and Escherichia coli. Virtual screenings based on molecular docking and the pharmacophore model, molecular dynamics simulations, and free energy calculations pointed to 1,8-dihydroxy-4,5-dinitroanthraquinone (DHDNA) as the most promising inhibitor. In addition, these analyses highlighted the contribution of the nitro group to the affinity of this anthraquinone for the nucleotide-binding site of PPAT. Furthermore, DHDNA was active in vitro towards Gram-positive bacteria with minimum inhibitory concentration (MIC) values of 31.25 µg/mL for S. aureus and 62.5 µg/mL for E. faecalis against both antibiotic-resistant isolates and reference strains but was ineffective against E. coli. Experiments on kill-time kinetics indicated that, at the tested concentrations, DHDNA produced bacteriostatic effects on both Gram-positive bacteria. Overall, our results present DHDNA as a potential PPAT inhibitor, showing antibacterial activity against antibiotic-resistant isolates of S. aureus and E. faecalis, findings that point to nitro groups as key to explaining these results.

The influence of different culture media on Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus biofilm formation

Microorganisms such as Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus are frequently isolated in samples of urinary, blood, intestinal, and respiratory infections, among others. These bacteria are also associated with microbial biofilm formation. E. coli, P. aeruginosa, and S. aureus biofilm infections are particularly hard to manage and often associated with nosocomial problems. This study investigated the influence of different culture media on E. coli, P. aeruginosa, and S. aureus biofilm formation. Bacterial performance was evaluated in brain heart infusion broth, Mueller-Hinton broth, or tryptic soy broth, with or without supplementing with different glucose levels (1-5%). The study quantified biofilm biomass and the count of viable biofilm colonies. This is the first study that compares the biofilm formation of E. coli, P. aeruginosa and S. aureus in polystyrene using different culture media and with different glucose concentrations. The most robust growth of E. coli, P. aeruginosa, and S. aureus occurred in brain heart infusion broth supplemented with 5% glucose, Mueller-Hinton broth without glucose, and tryptic soy broth with 2% glucose, respectively. Our data demonstrate that behavioral and morphological characteristics of each bacterium require a specific broth to enhance the growth of these microorganisms. These findings will contribute to future tests for therapeutic alternatives with anti-biofilm potential.

Corrigendum: Identification of novel thermosensors in gram-positive pathogens.

[This corrects the article DOI: 10.3389/fmolb.2020.592747.].