ABSTRACT
Thechemical composition, antioxidant and antimicrobial activities of the essential oil from aerial parts (leaves and flowers) of Chuquiraga arcuataHarling grown in the Ecuadorian Andes were studied. One hundred and twenty-six compounds were identified in the essential oil. Monoterpene hydrocarbons (45.8%) and oxygenated monoterpenes (44.1%) had the major percentages. The most abundant compounds were camphor (21.6%), myrcene (19.5%), and 1,8-cineole (13.4%). Antioxidant activity was examined using DPPH, ABTS,and FRAP assays. The essential oil had a moderate scavenging effect and reduction of ferric ion capacity through FRAP assay. Antimicrobial activity of the essential oil was observed against four pathogenic bacteria and a fungus. The essential oil exhibited activity against all microorganism strains under test, particularly against Candida albicansand Staphylococcus aureuswith MICs of 2.43-12.10 µg/mL.
Se estudió la composición química, actividades antioxidantes y antimicrobianas del aceite esencial procedente de las partes aérea (hojas y flores) de Chuquiraga arcuataHarling cultivadas en los Andes ecuatorianos. Se identificaron 126 compuestos en el aceite esencial. Los hidrocarburos monoterpénicos (45,8%) y los monoterpenos oxigenados (44,1%) tuvieron el mayor porcentaje. Los compuestos más abundantes fueron alcanfor (21,6%), mirceno (19,5%) y 1,8-cineol (13,4%). La actividadantioxidante se examinó mediante ensayos DPPH, ABTS y FRAP. El aceite esencial tuvo un efecto eliminador moderado y una reducción de la capacidad de iones férricos mediante el ensayo FRAP. Se observó actividad antimicrobiana del aceite esencial contra cuatro bacterias y un hongo patógenos. El aceite esencial mostró actividad contra todas las cepas de microorganismos bajo prueba, particularmente contra Candida albicansy Staphylococcus aureuscon CMI de 2,43-12,10 µg/mL.
Subject(s)
Oils, Volatile/chemistry , Plant Extracts/chemistry , Asteraceae/chemistry , Anti-Infective Agents/chemistry , Antioxidants/chemistry , Bacteria/drug effects , Oils, Volatile/pharmacology , Plant Extracts/pharmacology , Chromatography, Gas , Plant Leaves/chemistry , Monoterpenes/analysis , Ecuador , Hydrocarbons/analysis , Anti-Infective Agents/pharmacology , Antioxidants/pharmacologyABSTRACT
The aim of this study was to optimize by response surface design, the extraction of the leaf essential oil (EO) from Minthostachys mollis [HBK] Griseb., grown in Ecuador, using steam distillation. The factors used were extraction time (XTIE) of 60, 105 and 150 min and plant material/water ratio (XRMA) of 1:3, 1:4 and 1:5. The optimal combination was reached with XRMA 1:5 and XTIE 150 min, obtaining a process yield of 0.67%. The chemical composition of the EO analyzed by GC - MS was determined, where the main compounds were carvacryl acetate (44.01%), carvacrol (16.51%) and menthone (8.20%). The anti oxidant capacity of EO was evaluated using the FRAP and ABTS methodologies, with an IC 50 243.21 µmol Fe 2+ /g and 0.12 mg/mL, respectively. In addition, the antimicrobial activity of EO was found against Pseudomonas aeruginosa , Salmonella enterica , Escherich ia coli and Staphylococcus aureus .
El objetivo del estudio fue optimizar, mediante un diseño de superficie respuesta, la extracción d el aceite esencial (AE) de hojas de Minthostachys mollis [HBK] Griseb. del Ecuador, mediante destilación por arrastre de vapor. Los factores fueron el tiempo de extracción (XTIE) de 60, 105 y 150 min, y relación de material vegetal/ agua destilada (XRMA) d e 1:3, 1:4 y 1:5. La combinación óptima se logró con XTIE 150 min y XRMA 1:5 para un rendimiento de 0,67%. Se determinó la composición química del AE por GC - MS donde los compuestos mayoritarios fueron acetato de carvacrilo (44,01%), carvacrol (16,51%) y me ntona (8,20%). Se evaluó la capacidad antioxidante del AE por las metodologías FRAP y ABTS, con CI 50 de 243,21 µmol Fe 2+ /g y 0,12 mg/mL, respectivamente. Además, se demostró la actividad antimicrobiana contra Pseudomonas aeruginosa , Salmonella enterica , Es cherichia coli y Staphylococcus aureus .
Subject(s)
Oils, Volatile/chemistry , Lamiaceae/chemistry , Anti-Bacterial Agents/chemistry , Antioxidants/chemistry , Bacteria/drug effects , Oils, Volatile/pharmacology , Plant Leaves , Ecuador , Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacologyABSTRACT
Introduction: Microorganism infiltration through the im-plant-abutment interface causes oral health problems such as periimplantitis, leading to implant loss. Materials and Methods: A feasible new method to quantify the Streptococcus mutans (S. mutans) infiltration through the implant-abutment interface gap is introduced in the present work. Internal hexagon (IH; n = 10), external hexagon (EH; n = 10), Morse taper (MT; n = 10), and a control for each group (n = 1) were tested. Bacteria suspension was prepared at 1.5x108 CFU/mL (CFU: colony forming units), and the implants were individually submerged up to the connection level, allowing the bacteria to contact it. The abutment was removed, and bacteria count was performed. Results: The implant sets were tested under normal bacterial growth and early and late biofilm growth conditions. Colony-forming units per mL were obtained, and the results were compared among groups. Differences in bacterial count between the MT and EH (p<0.001) and the MT and IH (p<0.001) groups were significantly higher in the MT-type implant. There was a significant increment of bacterial infiltration in the MTs submitted to late biofilm growth conditions. EH and IH connections are more effective in preventing bacterial infiltration independent of the growth condition. Conclusions: The proposed methodology is feasible to evaluate the infiltration of microorganisms through the implant-abutment interface.
Introducción: La infiltración de microorganismos a través de la interfaz implante-pilar provoca problemas de salud bucal como la periimplantitis, que conduce a la pérdida del implante. Materiales y Métodos: En el presente trabajo se presenta un nuevo método factible para cuantificar la infiltración de Streptococcus mutans (S. mutans) a través de la brecha de la interfaz implante-pilar. Se probaron el hexágono interno (IH; n = 10), el hexágono externo (EH; n = 10), el cono Morse (MT; n = 10) y un control para cada grupo (n = 1). Se preparó una suspensión de bacterias a 1,5x108 UFC/mL y los implantes se sumergieron individualmente hasta el nivel de conexión, permitiendo que las bacterias entraran en contacto con él. Resultados: Se retiró el pilar y se realizó recuento de bacterias. Los conjuntos de implantes se probaron en condiciones de crecimiento bacteriano normal y de crecimiento temprano y tardío de biopelículas. Se obtuvieron unidades formadoras de colonias por ml y los resultados se compararon entre grupos. Las diferencias en el recuento bacteriano entre los grupos MT y EH (p<0,001) y MT e IH (p<0,001) fueron significativamente mayores en el implante tipo MT. Hubo un incremento significativo de la infiltración bacteriana en los MT sometidos a condiciones tardías de crecimiento de biopelículas. Las conexiones EH e IH son más efectivas para prevenir la infiltración bacteriana independientemente de las condiciones de crecimiento. Conclusión: La metodología propuesta es factible para evaluar la infiltración de microorganismos a través de la interfaz implante-pilar.
Subject(s)
Humans , Dental Implants/microbiology , Dental Abutments/microbiology , Dental Leakage/microbiology , Dental Leakage/prevention & control , Streptococcus mutans/isolation & purification , Bacteria , BiofilmsABSTRACT
Abstract The objective of this research was to identify bacteria present in the microbiota of dentinal carious lesions in primary molars of some Costa Rican pediatric patients. Data were collected from 15 children aged between 4 and 8 years old who attended the Pediatric Dentistry Clinic at the Faculty of Dentistry from the University of Costa Rica (UCR). The inclusion criteria were: infants between 4 and 8 years old who presented cavitated carious lesions in primary teeth, who were actively attended by students at the Faculty of Dentistry from the UCR, and whose parents or legal guardians signed the informed consent to participate in this research. Samples were taken using a sterile spoon, placed in storage vials, and subjected to various conventional and molecular microbial identification techniques, such as Gram stain identification, catalase tests, oxidase, TSI, API 20E, API STAPH, and VITEK 2. Of the 60 bacterial strains subjected to Gram staining, the following was obtained: 28 Gram-positive bacteria and 32 Gram-negative bacteria. The main isolated organisms were species of Staphylococcus epidermidis, Pasteurella pneumotropica/Mannheimia haemolytica, Pantoea spp, and Streptococcus mutans.
Resumen El objetivo de esta investigación fue identificar las bacterias presentes en la microbiota de lesiones cariosas dentinales en molares primarias de pacientes pediátricos costarricenses. Las muestras fueron recolectadas de 15 niños entre los 4 y 8 años que fueron atendidos en la Clínica de Odontopediatría en la Facultad de Odontología de la Universidad de Costa Rica (UCR). Los criterios de inclusión fueron: pacientes entre los 4 y los 8 años de edad que presentaran lesiones cariosas cavitadas en dientes primarios, que se encuentraran activos para su atención por estudiantes de la Facultad de Odontología de la UCR, y que los padres o encargados legales firmaran el consentimiento informado para participar en esta investigación. Las muestras se tomaron utilizando una cuchareta estéril, colocándolas en viales de almacenamiento y fueron sometidas a diversas técnicas de identificación microbiana convencionales y moleculares tales como: identificación por Tinción de Gram, pruebas catalasa, oxidasa, TSI, API 20E, API STAPH y VITEK. De las 60 cepas bacterianas sometidas a tinción de Gram se obtuvo: 28 bacterias Gram Positivas y 32 bacterias Gram Negativas. Los principales organismos aislados fueron: especies de Staphylococcus epidermidis, Pasteurella pneumotropica/ Mannheimia haemolytica, Pantoea spp y Streptococcus mutans.
Subject(s)
Humans , Male , Female , Child, Preschool , Child , Bacteria/classification , Dental Caries/diagnosis , Microbiota , Tooth, Deciduous/microbiology , Costa RicaABSTRACT
Abstract Introduction: Sea urchin diseases have been documented in several locations worldwide, with reported occurrences of bacterial, protozoan, fungal, and algal infections. Objective: This study aimed to investigate pathogen agents in populations of Arbacia lixula and Paracentrotus lividus along the coast of Gran Canaria Island (Central-East Atlantic, Spain). Methods: Sampling was conducted at San Cristobal beach, on the Northeast side of the island, where sea urchins were manually collected from depths of 1-3 m during June, July, and October 2022. Swab samples were taken from the external and internal areas of the lesions and cultured on various media plates. Results: Eight different pathogen agents, including bacteria and fungi, were identified, with Vibrio alginolyticus being the most frequently observed bacteria in all diseased sea urchin samples. Additionally, ciliated protozoans were found within the tests, potentially acting as opportunistic parasites. Conclusions: This research provides a unique perspective on bald sea urchin disease by identifying a significant number of associated pathogens, including Candida, previously unreported in diseased organisms. Furthermore, the study highlights the presence of an inflammatory response in tissues with bacterial colonies, offering crucial insights into understanding this sea urchin disease.
Resumen Introducción: Las enfermedades en los erizos de mar han sido descritas en muchas localidades alrededor del mundo, y se han asociado con la presencia de infecciones por bacterias, protozoarios, hongos y algas. Objetivo: Este estudio tuvo como finalidad investigar sobre los agentes patógenos que afectan a las poblaciones de Arbacia lixula y Paracentrotus lividus a lo largo de la costa de la Isla de Gran Canaria (Atlántico Centro-Oriental, España). Métodos: El muestreo fue llevado a cabo en la playa de San Cristóbal, al noreste de la isla, dónde los organismos fueron capturados entre 1-3 metros de profundidad, durante junio, julio y octubre del año 2022. Se tomaron muestras en la zona interna y externa de la lesión en cada organismo, y se cultivaron en varios medios de cultivo. Resultados: Fueron identificados ocho agentes patógenos diferentes, incluyendo bacterias y hongos, y siendo Vibrio alginolyticus la bacteria más frecuentemente observada en todas las muestras de erizos enfermos. Además, se observaron protozoarios ciliados dentro de los caparazones, actuando potencialmente como parásitos oportunistas. Conclusiones: Esta investigación proporciona una perspectiva única sobre la enfermedad del erizo desnudo al identificar un número significativo de patógenos asociados, incluida Candida, que no se había reportado previamente en organismos enfermos. Además, el estudio destaca la presencia de una respuesta inflamatoria en tejidos con colonias bacterianas, lo que ofrece información crucial para comprender esta enfermedad de los erizos de mar.
Subject(s)
Animals , Bacteria , Arbacia/pathogenicity , FungiABSTRACT
Although numerous studies have demonstrated the biomedical potential of Myrtus communis L., (Myrtaceae) data on myrt le from Montenegro are scarce. T o evaluate antioxidant, antimutagenic and antibacterial activity of myrtle methanolic extracts. Antioxidant activity was evaluated by measuring free radicals scavenging activity, reducing power and enzyme inhibition. The strongest scavenging activity was towards DPPH radical ( 2,2 - diphenyl - 1 - picry lhydrazyl) (IC 50 1.69 - 2.25 mg/mL) and superoxide anion (IC 50 0.56 to 0.88 mg/mL), followed by high reducing power (428 - 472 mgAA/g.DE) and inhibition of XOD (IC 50 0.308 - 0.6261mg/mL). Antimutagenic activity was evaluated in reverse mutation assays with Esche richia coli WP2 oxyR mutant IC202 and deficient in the induction of antioxidant enzymes. The myrtle extracts strongly inhibited mutagenesis induced by t - BOOH, reaching 70% at the highest concentration applied. Antimicrobial activity was examined on eight different bacterial strains. Gram - positive bacteria, S. epidermis , S. aureus and M. flavus demonstrated the highest sensitivity towards extracts (MICs 4.5 - 9 mg/mL), but significantly lower towards essential oil (MIC 0.42 - 3.32 mg/mL).
Aunque numerosos estudios han demostrado el potencial biomédico de Myrtus communis L., (Myrtaceae), los datos sobre el mirto de Montenegro son escasos. E valuar la actividad antioxidante, antimutagéni ca y antibacteriana de extractos metanólicos de mirto. La actividad antioxidante se evaluó midiendo la actividad de eliminación de radicales libres, el poder reductor y la inhibición enzimática. La actividad secuestrante más fuerte fue hacia DPPH radical ( IC 50 1.69 - 2.25 mg/mL) y radicales de anión superóxido (IC 50 0.56 a 0.88 mg/mL), seguido de alto poder reductor (428 - 472 m gAA/g.DE) e inhibición de XOD (I C 50 0,308 - 0,6261 mg/m L ). La actividad antimutagénica se evaluó en ensayos de mutación inversa con Esche richia coli WP2 oxyR mutante IC202 y deficiente en la inducción de enzimas antioxidantes. Los extractos de mirto inhibieron fuertemente la mutagénesis inducida por t - BOOH, alcanzando el 70% a la mayor concentración aplicada. La actividad antimicrobiana se examinó en octo cepas bacterianas diferentes. Las bacterias grampositivas, S. epidermis , S. aureus y M. flavus demostraron la sensibilidad más alta hacia los extractos (MIC 4.5 - 9 mg/mL), pero significativamente más baja hacia el aceite esencial (MIC 0.42 - 3 .32 mg/mL). Los resultados muestran la gran perspectiva nutrafarmacéutica de la especie montenegrina Myrtus communis .
Subject(s)
Oils, Volatile/pharmacology , Plant Extracts/pharmacology , Myrtus/chemistry , Phenols/analysis , Bacteria/drug effects , Flavonoids/analysis , Oils, Volatile/chemistry , Plant Extracts/chemistry , Antimutagenic Agents/pharmacology , Montenegro , Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacologyABSTRACT
BACKGROUND@#Links between alterations in gut microbiota composition and amyotrophic lateral sclerosis (ALS) have previously been reported. This study aimed to examine the microbiota in the nasal cavity of ALS.@*METHODS@#Sixty-six ALS patients and 40 healthy caregivers who live in close proximity with patients were enrolled. High throughput metagenomic sequencing of the 16S ribosomal deoxyribonucleic acid (rDNA) gene V3-V4 region of nasal microbiota was used to characterize the alpha and beta diversity and relative abundance of bacterial taxa, predict function, and conduct correlation analysis between specific taxa and clinical features.@*RESULTS@#The nasal microbiome of ALS patients showed lower alpha diversity than that of corresponding healthy family members. Genera Gaiella , Sphingomonas , Polaribacter _1, Lachnospiraceae _NK4A136_group, Klebsiella , and Alistipes were differentially enriched in ALS patients compared to controls. Nasal microbiota composition in ALS patients significantly differed from that in healthy subjects (unweighted UniFrac P = 0.001), while Linear discriminant analysis Effect Size (LEfSe) analysis indicated that Bacteroidetes and Firmicutes dominated healthy nasal communities at the phylum level, whereas Actinobacteria was the predominant phylum and Thermoleophilia was the predominant class in ALS patients. Genus Faecalibacterium and Alistipes were positively correlated with ALS functional rating scale revised (ALSFRS-R; rs = 0.349, P = 0.020 and rs = 0.393, P = 0.008), while Prevotella -9 and Bacteroides operational taxonomic units (OTUs) were positively associated with lung function (FVC) in ALS patients ( rs = 0.304, P = 0.045, and rs = 0.300, P = 0.048, respectively). Prevotella -1 was positively correlated with white blood cell counts (WBC, rs = 0.347, P = 0.021), neutrophil percentage (Neu%, rs = 0.428, P = 0.004), and neutrophil-to-lymphocyte ratio (NLR, rs = 0.411, P = 0.006), but negatively correlated with lymphocyte percentage (Lym%, rs = -0.408, P = 0.006). In contrast, Streptococcus was negatively associated with Neu% ( rs = -0.445, P = 0.003) and NLR ( rs = -0.436, P = 0.003), while positively associated with Lym% ( rs = 0.437, P = 0.003). No significant differences in nasal microbiota richness and evenness were detected among the severe and mild ALS patients.@*CONCLUSIONS@#ALS is accompanied by altered nasal microbial community composition and diversity. The findings presented here highlight the need to understand how dysbiosis of nasal microbiota may contribute to the development of ALS.
Subject(s)
Humans , Amyotrophic Lateral Sclerosis/microbiology , Feces/microbiology , Microbiota/genetics , Gastrointestinal Microbiome/genetics , Bacteria/genetics , RNA, Ribosomal, 16S/geneticsABSTRACT
OBJECTIVE@#The aim of this study was to assess the impact of bisphenol A (BPA) and its substitute, bisphenol F (BPF), on the colonic fecal community structure and function of mice.@*METHODS@#We exposed 6-8-week-old male C57BL/6 mice to 5 mg/(kg∙day) and 50 μg/(kg∙day) of BPA or BPF for 14 days. Fecal samples from the colon were analyzed using 16S rRNA sequencing.@*RESULTS@#Gut microbiome community richness and diversity, species composition, and function were significantly altered in mice exposed to BPA or BPF. This change was characterized by elevated levels of Ruminococcaceae UCG-010 and Oscillibacter and decreased levels of Prevotella 9 and Streptococcus. Additionally, pathways related to carbohydrate and amino acid metabolism showed substantial enrichment.@*CONCLUSION@#Mice exposed to different BP analogs exhibited distinct gut bacterial community richness, composition, and related metabolic pathways. Considering the essential role of gut bacteria in maintaining intestinal homeostasis, our study highlights the intestinal toxicity of BPs in vertebrates.
Subject(s)
Male , Animals , Mice , Gastrointestinal Microbiome , Mice, Inbred C57BL , RNA, Ribosomal, 16S/genetics , Benzhydryl Compounds/toxicity , Bacteria/genetics , PhenolsABSTRACT
Due to the advancement of 16S rRNA sequencing technology, the lower respiratory tract microbiota, which was considered non-existent, has been revealed. The correlation between these microorganisms and diseases such as tumor has been a hot topic in recent years. As the bacteria in the surrounding can infiltrate the tumors, researchers have also begun to pay attention to the biological behavior of tumor bacteria and their interaction with tumors. In this review, we present the characteristic of the lower respiratory tract bacteria and summarize recent research findings on the relationship between these microbiota and lung cancer. On top of that, we also summarize the basic feature of bacteria in tumors and focus on the characteristic of the bacteria in lung cancer. The relationship between bacteria in lung cancer and tumor development is also been discussed. Finally, we review the potential clinical applications of bacterial communities in the lower respiratory tract and lung cancer, and summarize key points of sample collection, sequencing, and contamination control, hoping to provide new ideas for the screening and treatment of tumors. .
Subject(s)
Humans , Lung Neoplasms , RNA, Ribosomal, 16S/genetics , Bacteria/genetics , Microbiota , Respiratory System , Lung/microbiologyABSTRACT
The reduction of nitrate to nitrite by the oral microbiota has been proposed to be important for oral health and results in nitric oxide formation that can improve cardiometabolic conditions. Studies of bacterial composition in subgingival plaque suggest that nitrate-reducing bacteria are associated with periodontal health, but the impact of periodontitis on nitrate-reducing capacity (NRC) and, therefore, nitric oxide availability has not been evaluated. The current study aimed to evaluate how periodontitis affects the NRC of the oral microbiota. First, 16S rRNA sequencing data from five different countries were analyzed, revealing that nitrate-reducing bacteria were significantly lower in subgingival plaque of periodontitis patients compared with healthy individuals (P < 0.05 in all five datasets with n = 20-82 samples per dataset). Secondly, subgingival plaque, saliva, and plasma samples were obtained from 42 periodontitis patients before and after periodontal treatment. The oral NRC was determined in vitro by incubating saliva with 8 mmol/L nitrate (a concentration found in saliva after nitrate-rich vegetable intake) and compared with the NRC of 15 healthy individuals. Salivary NRC was found to be diminished in periodontal patients before treatment (P < 0.05) but recovered to healthy levels 90 days post-treatment. Additionally, the subgingival levels of nitrate-reducing bacteria increased after treatment and correlated negatively with periodontitis-associated bacteria (P < 0.01). No significant effect of periodontal treatment on the baseline saliva and plasma nitrate and nitrite levels was found, indicating that differences in the NRC may only be revealed after nitrate intake. Our results suggest that an impaired NRC in periodontitis could limit dietary nitrate-derived nitric oxide levels, and the effect on systemic health should be explored in future studies.
Subject(s)
Humans , Nitrates , Nitric Oxide , Nitrites , RNA, Ribosomal, 16S/genetics , Periodontitis/microbiology , Bacteria , Dental Plaque/microbiology , Saliva/microbiology , Microbiota/geneticsABSTRACT
Abstract Fish is the main source of animal protein for human diet. The aim of this study was to find out prevalence of pathogenic bacteria of two selected economically important fish of Pakistan namely Mahseer (Tor putitora) and Silver carp (Hypophthalmichthys molitrix). Live fish samples from hatcheries and dead fish samples from different markets of study area were randomly collected. The fish samples were analyzed for isolation, identification and prevalence of bacteria. The isolated bacteria from study fish were identified through biochemical test and about 10 species of pathogenic bacteria were identified including the pathogenic bacteria to human and fish namely, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. and Salmonella spp. The bacterial percentage frequency of occurrence in Silver carp and Mahseer fish showed Pseudomonas aeruginosa 21.42%, Staphylococcus epidermidis 17.85%, Escherichia coli 11.90%, Staphylococcus aureus 9.52%, Citrobacter spp. 9.52%, Serratia spp. 8.33%, Streptococcus iniae 7.14%, Stenotrophomonas spp. 5.95%, Bacillus spp. 4.76% and Salmonella spp. 3.57%. The study revealed that Fish samples of Mahseer and Silver carp that were collected from markets have found more isolates (10 bacterial species) than did the fresh fish pond samples (03 bacterial species) of hatcheries. The occurrence of pathogenic bacteria in study fish showed risk factor for public health consumers.
Resumo O peixe é a principal fonte de proteína animal para a alimentação humana. O objetivo deste estudo foi descobrir a prevalência de bactérias patogênicas de dois peixes economicamente importantes selecionados do Paquistão, nomeadamente Mahseer (Tor putitora) e carpa prateada (Hypophthalmichthys molitrix). Amostras de peixes vivos de incubatórios e amostras de peixes mortos de diferentes mercados da área de estudo foram coletadas aleatoriamente. As amostras de peixes foram analisadas quanto ao isolamento, identificação e prevalência de bactérias. As bactérias isoladas dos peixes do estudo foram identificadas através de testes bioquímicos e cerca de 10 espécies de bactérias patogênicas foram identificadas incluindo as bactérias patogênicas para humanos e peixes, nomeadamente, Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus iniae, Serratia spp. Citrobacter spp. Stenotrophomonas spp. Bacillus spp. e Salmonella spp. A porcentagem de freqüência de ocorrência bacteriana em carpa prateada e peixes Mahseer mostrou Pseudomonas aeruginosa 21,42%, Staphylococcus epidermidis 17,85%, Escherichia coli 11,90%, Staphylococcus aureus 9,52%, Citrobacter spp. 9,52%, Serratia spp. 8,33%, Streptococcus iniae 7,14%, Stenotrophomonas spp. 5,95%, Bacillus spp. 4,76% e Salmonella spp. 3,57%. O estudo revelou que as amostras de peixes de Mahseer e carpa prateada coletadas nos mercados encontraram mais isolados (10 espécies bacterianas) do que as amostras de peixes frescos (03 espécies bacterianas) de incubatórios. A ocorrência de bactérias patogênicas nos peixes do estudo apresentou fator de risco para consumidores de saúde pública.
Subject(s)
Humans , Animals , Carps , Pakistan , Bacteria , Ponds , IncidenceABSTRACT
Abstract The impact of antibiotics on growth, cocoon production was assessed in addition to isolation and characterization of bacteria associated with silkworm gut of infected larvae. Larval rearing was maintained at recommended conditions of temperature and humidity. Silkworm larvae showing abnormal symptoms were collected from the control group and dissected for gut collection. Bacteria were isolated from the gut content by spreading on agar plates and incubated at 37 °C for 48 hrs. Bacterial identification and phylogenetic analysis were carried out by 16S rRNA gene sequencing. The isolated bacteria were subjected to antimicrobial susceptibility test (disc diffusion methods) by using Penicillin (10 µg/mL), Tetracycline (30 µg/mL), Amoxicillin (25 µg/mL), Ampicillin (10 µg/mL), and Erythromycin (15 µg/mL). All isolated strains showed positive results for the catalase test. We isolated and identified bacterial strains (n = 06) from the gut of healthy and diseased silkworm larvae. Based on the 16S rRNA gene sequence, isolated bacteria showed close relation with Serratia, Bacillus, and Pseudomonas spp. Notably, 83.3% of strains were resistant to Penicillin, Tetracycline, Amoxicillin, Ampicillin, and Erythromycin but 16.6% showed antibiotic susceptibility to the above-mentioned commonly used antibiotics. Silkworm larvae fed on penicillin-treated leaves showed significant improvement in larval weight, larval length, and cocoon production. Significantly higher larval weight (6.88g), larval length (5.84cm), and cocoon weight (1.33g) were recorded for larvae fed on leaves treated with penicillin as compared to other antibiotics. Isolated bacterial strains showed close relation with Serratia spp., Bacillus spp. and Pseudomonas spp.
Resumo O impacto dos antibióticos no crescimento e na produção do casulo foi avaliado, além do isolamento e caracterização das bactérias associadas ao intestino de larvas infectadas do bicho-da-seda. A criação das larvas foi mantida nas condições recomendadas de temperatura e umidade. As larvas do bicho-da-seda com sintomas anormais foram coletadas do grupo controle e dissecadas para coleta do intestino. As bactérias foram isoladas do conteúdo intestinal por espalhamento em placas de ágar e incubadas a 37° C durante 48 horas. A identificação bacteriana e a análise filogenética foram realizadas pelo sequenciamento do gene 16S rRNA. As bactérias isoladas foram submetidas a teste de sensibilidade antimicrobiana (métodos de difusão em disco) com penicilina (10 µg / mL), tetraciclina (30 µg / mL), amoxicilina (25 µg / mL), ampicilina (10 µg / mL) e eritromicina (15 µg / mL). Todas as cepas isoladas apresentaram resultados positivos para o teste da catalase. Isolamos e identificamos cepas bacterianas (n = 06) do intestino de larvas de bicho-da-seda saudáveis e doentes. Com base na sequência do gene 16S rRNA, as bactérias isoladas mostraram estreita relação com Serratia, Bacillus e Pseudomonas spp. Notavelmente, 83,3% das cepas eram resistentes a penicilina, tetraciclina, amoxicilina, ampicilina e eritromicina, mas 16,6% mostraram suscetibilidade aos antibióticos comumente usados mencionados acima. As larvas do bicho-da-seda alimentadas com folhas tratadas com penicilina apresentaram melhora significativa no peso larval, comprimento larval e produção de casulo. Peso larval significativamente maior (6,88g), comprimento larval (5,84cm) e peso do casulo (1,33g) foram registrados para larvas alimentadas com folhas tratadas com penicilina, em comparação com outros antibióticos. Cepas bacterianas isoladas mostraram estreita relação com Serratia spp., Bacillus spp. e Pseudomonas spp.
Subject(s)
Animals , Bombyx , Anti-Bacterial Agents/pharmacology , Phylogeny , Bacteria/genetics , RNA, Ribosomal, 16S/genetics , Microbial Sensitivity Tests , LarvaABSTRACT
Aim: This study aimed to perform an in vitro comparative analysis of the antifungal activity of different calcium silicate-based endodontic sealers against three fungal species. Methods: The antifungal properties of three calcium silicate-based sealers were tested: Bio-C Sealer, Cambiar a Sealer Plus BC, and MTA-Fillapex. Two commonly used sealers were used as controls: AH Plus and Endomethasone. An agar diffusion test was performed to analyze the antifungal activity of the sealers against Candida albicans, Candida glabrata, Candida tropicalis, and a mixed microbial culture medium. The results were analyzed using ANOVA (p <0.05). Results: Endomethasone exhibited the highest inhibition against all strains examined, maintaining a consistent level of inhibition throughout 7 days. MTA-Fillapex demonstrated the best performance among the calcium silicate-based sealers for the three fungal species (p < 0.05), maintaining stable values over the 7 days, surpassing that of Endomethasone. Nevertheless, MTA-Fillapex only exhibited antimicrobial effect against the mixed culture for the first 24 hours, and no antimicrobial activity was observed at 48 hours, being surpassed by all tested sealers (p < 0.05). Conclusion: Of all silicate-based sealers tested, only MTA-Fillapex exhibited promising antifungal activity. Nevertheless, care must be taken when extrapolating these results, as MTA-Fillapex exhibited poor antimicrobial activity when tested in mixed microbial cultures
Subject(s)
Root Canal Filling Materials , Silicate Cement , Bacteria , Candida albicans , Candida glabrata , Candida tropicalis , Endodontics , Antifungal Agents/analysisABSTRACT
Abstract For places where non-sterile drug production occurs, regulatory bodies recommend monitoring of the environmental bioburden. This procedure provides information regarding possible microbiological risks to which the products may be exposed, so that subsequent action measures may be implemented. The aim of the present work was to quantify and characterize the microorganisms present in Grade D (ISO 8) cleanrooms of a Brazilian pharmaceutical industry, identifying any possible seasonal climatic influences on these environments. Sampling was performed by surface and air monitoring, over 12 months during the year 2019, in rooms that were in operation. For both sampling methods, no statistically significant differences in bacteria and fungi counts were found between months or seasonal periods. Microorganisms that presented higher incidence included Staphylococcus epidermidis (15%) and Micrococcus spp. (13%), common to the human microbiota, and the fungi Cladosporium sp. (23%) and Penicillium sp. (21%), typical of the external environment. The results showed that microbial contamination in the Grade D cleanrooms was within the permissible maximum levels and remained similar throughout the year. Microbiological quality control in the clean areas of the pharmaceutical industry investigated was considered effective, with regular maintenance being necessary to keep bioburden levels controlled.
Subject(s)
Bacteria/classification , Environmental Monitoring/instrumentation , Environmental Monitoring/methods , Drug Industry/instrumentation , Environmental Pollution/prevention & control , Microbiota/immunology , Fungi/classificationABSTRACT
Aim: To investigate the bacteriological profile of oral and maxillofacial infections and the pattern of sensitivity to a specific group of antibiotics in a reference emergency hospital in Brazil. Methods: This is a prospective cohort institutional study that studied patients affected by oral and/ or maxillofacial infections in a Brazilian emergency hospital, over a 12-month period, of different etiologies, through data collection, culture and antibiogram tests, and monitoring of the process of resolution of the infectious condition. The variables were analyzed using the chi-square and Mann-Whitney tests, using a significance level of 5%. Results: The sample consisted of 61 patients, 62.3% male. The mean age of participants was 34.3 years. Odontogenic infection was the most frequent etiology and the submandibular space was the most affected. The bacterial species Streptococcus viridans was isolated in 21.6% of cases. Levofloxacin, vancomycin and penicillin were the antibiotics with the highest frequency of bacterial sensitivity, while clindamycin and erythromycin showed the highest percentages of resistance. Conclusions: The results suggest that, among the most used antibiotics for the treatment of these infections, penicillin remains an excellent option of choice for empirical therapy
Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Surgical Wound Infection , Bacteria/drug effects , Drug Resistance, Microbial , Focal Infection, Dental , Anti-Bacterial AgentsABSTRACT
RESUMO Objetivo: Avaliar o perfil clínico-epidemiológico dos pacientes e o perfil dos agentes causadores de ceratite infecciosa em pacientes submetidos à cultura obtida a partir de raspados corneanos em um serviço de emergência oftalmológica em um hospital de referência. Métodos: Análise retrospectiva de prontuários e resultados do exame de cultura de córnea. Os dados foram coletados após a aprovação do Comitê de Ética em Pesquisa com Seres Humanos. Resultados: A maioria dos pacientes era do sexo masculino, com média de idade de 47,6 anos, etnia caucasiana, baixo nível de escolaridade e procedentes da Grande Florianópolis. Trauma ocular foi o principal fator de risco observado. Foram obtidos resultados de cultura positivos em 43,6% dos casos, sendo Pseudomonas aeruginosa o principal agente identificado. O tratamento clínico foi a conduta inicial na maioria dos casos. Conclusão: Ceratite infecciosa é uma doença grave e o conhecimento atualizado é necessário para o diagnóstico precoce e manejo adequado da doença.
ABSTRACT Objective: To evaluate patients' clinical-epidemiological profile and causative agents of infectious keratitis on patients submitted to culture obtained from corneal smears in an ophthalmological emergency in a reference hospital. Methods: Retrospective analysis of medical records and corneal culture examination results. The data were collected after the approval of the Human Research Ethics Committee. Results: Most patients were male, with an average age of 47.6 years old, Caucasian ethnicity, low level of education, and from the Greater Florianópolis area. Ocular trauma was the main observed risk factor. Positive culture results were obtained in 43.6% of cases, with Pseudomonas aeruginosa being the main identified agent. Clinical treatment was the initial approach in most cases. Conclusion: Infectious keratitis is a serious disease, and up-to-date knowledge is necessary for early diagnosis and appropriate management of the disease.
Subject(s)
Humans , Male , Female , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged , Aged, 80 and over , Eye Infections/diagnosis , Eye Infections/etiology , Eye Infections/microbiology , Eye Infections/epidemiology , Keratitis/diagnosis , Keratitis/etiology , Keratitis/microbiology , Keratitis/epidemiology , Bacteria/isolation & purification , Acanthamoeba/isolation & purification , Eye Infections/therapy , Medical Records , Retrospective Studies , Risk Factors , Cornea/microbiology , Fungi/isolation & purification , Keratitis/therapyABSTRACT
INTRODUCCIÓN: Las infecciones bacterianas en trasplante hepático (TH) son una de las principales causas de morbimortalidad. OBJETIVO: Caracterizar las complicaciones infecciosas bacterianas en el primer mes postrasplante. Pacientes y MÉTODOS: Estudio retrospectivo entre los años 2009-2020. RESULTADOS: 225 pacientes recibieron un TH. 80 (35,5%) desarrollaron al menos un episodio de infección bacteriana en el primer mes postrasplante hepático. Hubo 105 episodios de infección bacteriana con una incidencia de 46,6%. El foco más frecuente fue el abdominal (48,6%) y el microorganismo predominante fue Klebsiella spp. De los 104 aislamientos, el 57,6% presentaron un perfil MDR/XDR. Los pacientes que desarrollaron una complicación infecciosa presentaron menor sobrevida al alta hospitalaria en comparación con los que no la presentaron 87,5 versus 94,5% [OR 4,18 (IC 95%: 1,5-11,6)]. En el análisis multivariado la reintervención quirúrgica precoz [OR 4,286 (IC 95%: 1,911-9,61)], mostró un riesgo significativo de desarrollar una complicación infecciosa bacteriana en el primer mes postrasplante. CONCLUSIONES: Tres de cada 10 pacientes presentaron una infección bacteriana en el primer mes postrasplante con una alta incidencia de bacilos gramnegativos MDR/XDR. Los pacientes que desarrollaron una complicación infecciosa presentaron una menor sobrevida al alta. La reintervención quirúrgica precoz se identificó como un factor predisponente de infección temprana.
BACKGROUND: Bacterial infections are one of the main causes of morbidity and mortality in liver transplant recipients (LT). Aim: To characterize bacterial infectious complications in the first month an after a liver transplant. METHODS: Retrospective analysis of a cohort of liver transplant recipients who presented at least one bacterial infectious complication in the first month after transplant between 2009 and 2020. RESULTS: 225 patients were analyzed. 80 (35.5%) had a least one documented bacterial infection during the first month after transplant. 105 bacterial infections were documented, with an incidence of 46.6%. The most frequent origin was intra-abdominal (48.6%) and the predominant isolated microorganism was Klebsiella spp. Among 104 isolated microorganisms 57.6% showed MDR/XDR profile. Patients who developed a bacterial infectious complication had a shorter overall survival (OS) after discharge from hospital (87.5% vs 94.5%) [OR 4.18 (IC 95%: 1.5-11,6)]. When multivariate analysis of predisposing factors was performed early surgical reoperation was the only variable associated with an increased risk of developing a bacterial complication in the first month [OR 4.286 (IC 95%: 1.911-9.61)]. CONCLUSIONS: Three out of 10 patients developed a bacterial infectious complication during the first month after liver transplant with a high incidence of gram-negative bacillus MDR/XDR. Patients who presented infectious complications had a shorter OS after discharge, and early reoperation was identified as a predisposing factor of early infectious complications.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Bacterial Infections/mortality , Liver Transplantation/adverse effects , Prognosis , Bacteria/isolation & purification , Bacterial Infections/etiology , Bacterial Infections/microbiology , Uruguay/epidemiology , Multivariate Analysis , Retrospective Studies , Risk FactorsABSTRACT
INTRODUCCIÓN: La diarrea aguda continúa siendo una de las principales causas de morbilidad en niños; sin embargo, el diagnóstico etiológico presenta limitaciones dada la baja sensibilidad de los métodos tradicionales. OBJETIVO: Describir los microorganismos identificados en niños que acudieron al Servicio de Urgencia (SU) de un hospital universitario en Santiago, Chile, por diarrea aguda y a los que se le solicitó panel molecular gastrointestinal. MÉTODOS: Se revisaron fichas clínicas y resultados de panel gastrointestinal realizados entre junio de 2017 y marzo de 2020. RESULTADOS: Se incluyeron 198 pacientes, edad promedio de 54,5 meses y 60,6% (120/198) de sexo masculino. La positividad del panel fue de 78,8% (156/198) con 35,3% (55/156) de las muestras polimicrobianas. Se identificaron 229 microorganismos, de los cuales 72,9% (167/229) corresponden a bacterias, 25,8% (59/229) a virus y 1,3% (3/229) a parásitos. Destacaron Campylobacter spp. y Escherichia coli enteropatógena (ECEP) como las bacterias más frecuentemente identificadas. Los pacientes con detección de Campylobacter spp. presentaron con mayor frecuencia fiebre (p = 0,00). ECEP se aisló principalmente (82,5%) en muestras polimicrobianas. DISCUSIÓN: Los resultados enfatizan el potencial que poseen los estudios moleculares para mejorar el diagnóstico etiológico de la diarrea, pero a la vez llevan a cuestionar el rol patogénico de algunos microorganismos identificados.
BACKGROUND: Acute diarrhea continues to be one of the main causes of morbidity in children, however the etiologica diagnosis presents limitations given the low sensitivity of traditional methods. AIM: To describe the microorganisms identified in children who attended the emergency department (ED) in Santiago, Chile, due to acute diarrhea and to whom a gastrointestinal panel was requested as part of their study. MATERIAL AND METHODS: Clinical records and results of the gastrointestinal panel carried out between June 2017 and March 2020 were reviewed. RESULTS: 198 patients were included, the average age was 54.5 months and 60.6% (120/198) were males. Positivity was 78.8% (156/198) with 35.3% (55/156) of the samples being polymicrobial. 229 microorganisms were identified, of which 72.9% (167/229) corresponded to bacteria, 25.8% (59/229) to viruses, and 1.3% (3/229) to parasites. Campylobacter spp. and enteropathogenic Escherichia coli (EPEC) were the most frequently identified bacteria. Patients with detection of Campylobacter spp. presented a higher frequency of fever (p = 0.00). EPEC was isolated in 82.5% of the cases in polymicrobial samples. DISCUSSION: The results emphasize the potential of molecular studies to improve the etiological diagnosis of diarrhea and at the same time lead to question the pathogenic role of some microorganisms.
Subject(s)
Humans , Male , Female , Diarrhea/diagnosis , Feces/microbiology , Parasites/isolation & purification , Seasons , Bacteria/isolation & purification , Viruses/isolation & purification , Chile , Retrospective Studies , Diarrhea/etiology , Diarrhea/epidemiology , Emergency Service, Hospital , Feces/parasitologyABSTRACT
INTRODUCCIÓN: Las opacidades pulmonares en receptores de trasplante de precursores hematopoyéticos (TPH) representan un desafío diagnóstico y son una causa de morbimortalidad. Existen grandes discrepancias con respecto a la sensibilidad diagnóstica del lavado broncoalveolar (LBA), sus complicaciones, y los factores asociados a la identificación microbiológica. OBJETIVO: Conocer la utilidad del estudio microbiológico del LBA en el diagnóstico, modificación de la conducta médica y estimar las complicaciones y mortalidad asociada al procedimiento, en receptores de TPH con opacidades pulmonares. PACIENTES Y MÉTODOS: Estudio de cohorte, retrospectivo, en adultos receptores de TPH a los que se les realizó una broncoscopía con LBA por presentar opacidades pulmonares, en el Hospital Italiano de Buenos Aires entre el 01/01/2011 y el 31/12/2020. RESULTADOS: De los 189 procedimientos analizados, en 79 se logró un hallazgo microbiológico (41,8%) y 122 permitieron modificar la conducta médica (64,6%). En 11 casos se observaron complicaciones graves dentro de las 12 horas (5,8%) de efectuado el LBA. La mortalidad intrahospitalaria fue de 16,8% (N = 21/125). El valor de neutrófilos en sangre previo al LBA (p = 0,037) y la presencia de nódulos pulmonares como lesión tomográfica predominante (p = 0,029) se asociaron independientemente al hallazgo microbiològico global. CONCLUSIONES: Nuestra investigación apoya la realización del LBA como herramienta diagnóstica en pacientes que reciben un TPH y presentan opacidades pulmonares.
BACKGROUND: Lung opacities are a cause of morbimortality in bone marrow transplant patients, and represent a diagnostic challenge. There are large discrepancies regarding the diagnostic sensitivity of bronchoalveolar lavage (BAL), its complications, and the factors associated with microbiological detection. AIM: To know the usefulness of the microbiological study of BAL in the diagnosis, in the modification in medical behavior and to estimate the complications and associated mortality of this diagnostic procedure in patients transplanted with hematopoietic progenitor cells with pulmonary opacities. METHODS: Retrospective cohort study in bone marrow transplant adult patients who underwent bronchoscopy with BAL due to lung opacities at Hospital Italiano de Buenos Aires between 01/01/2011 and 12/31/2020. RESULTS: Of the 189 BAL analyzed, 79 presented a microbiological detection (41.8%) and 122 allowed to modify the medical behavior (64.6%). Severe complications were observed within 12 hours after the procedure in11 cases (5.8%). In-hospital mortality was 16,8% (N = 21/125). The value of blood neutrophils prior to bronchoalveolar lavage (p = 0.037) and the presence of pulmonary nodules as the predominant tomographic lesion (p = 0.029) were independently associated with global microbiological detection. CONCLUSION: Our research supports the performance of BAL as a diagnostic tool in bone marrow transplant patients with lung opacities.
Subject(s)
Humans , Male , Female , Middle Aged , Bronchoscopy/methods , Bronchoalveolar Lavage Fluid/microbiology , Hematopoietic Stem Cell Transplantation/adverse effects , Bronchoalveolar Lavage/methods , Hematologic Neoplasms/therapy , Bacteria/isolation & purification , Viruses/isolation & purification , Multivariate Analysis , Cohort Studies , Immunocompromised Host , Transplant Recipients , Fungi/isolation & purification , Lung/microbiologyABSTRACT
INTRODUCCIÓN: El absceso renal es infrecuente en pediatría, con clínica y laboratorio inespecíficos. Ante su sospecha, es necesario realizar imágenes para establecer diagnóstico. Objetivo: Describir las características clínico-epidemiológicas, microbiológicas, diagnósticas y terapéuticas de abscesos renales en pediatría. PACIENTES Y MÉTODOS: Estudio retrospectivo, descriptivo, de pacientes internados con absceso renal, en seguimiento por Infectología del Hospital de Niños Ricardo Gutiérrez, durante 9 años. RESULTADOS: 15 pacientes (67% varones), mediana de edad 9 años (rango [r] 0,7-17). Cuatro pacientes con comorbilidades. El síntoma más frecuente fue fiebre seguido por dolor lumbar. El recuento medio de leucocitos en sangre fue de 15.700/mm3 (r: 7.100-45.000) y la PCR de 193 mg/L (r: 1-362). Cuatro pacientes presentaron urocultivo positivo: dos Escherichia coli, uno Klebsiella pneumoniae y E. coli y otro Candida albicans y K. pneumoniae. Ningún paciente presentó bacteriemia. El diagnóstico se confirmó por ecografía. Se realizó drenaje en siete pacientes, con aislamiento de Staphylococcus aureus en dos y Pseudomonas aeruginosa en uno. El tratamiento incluyó terapia combinada en 67%. Mediana de antibioterapia intravenosa fue 16 días (r: 7-49), total de 28 (r: 14-91). Un paciente requirió terapia intensiva y dos, nefrectomía. CONCLUSIÓN: Los abscesos renales son infrecuentes, con gran morbimortalidad. Sospechar en paciente con infección del tracto urinario (ITU) de evolución tórpida que persiste febril. En nuestro estudio, la alta sensibilidad de la ecografía renal permitió su diagnóstico precoz.
BACKGROUND: Renal abscesses are infrequent in pediatrics, with nonspecific clinical and laboratory findings. When suspected, imaging is essential to establish the diagnosis. Aim: To describe the clinical-epidemiological, microbiological, diagnostic and therapeutic characteristics of renal abscesses in pediatrics. METHODS: Retrospective and descriptive study of hospitalized patients with renal abscess, followed by Infectious Diseases Department of Ricardo Gutiérrez Children's Hospital during 9 years. Statistical analysis: Epi Info 7.2.2.6. RESULTS: 15 patients (67% male), median age 9 years (range [r] 0.7-17) were included. Four patients had underlying disease. The most frequent symptom was fever, with a median duration of 10 days (r:1-36), followed by lumbar pain. The median white blood cell count was 15,700/mm3 (r: 7,100-45,000) and CRP 193mg/L (r: 1-362). Four patients presented positive urine culture: 2 Escherichia coli, 1 Klebsiella pneumoniae and E. coli and 1 Candida albicans and K. pneumoniae. No patient had bacteremia. The diagnosis of abscess was confirmed by ultrasound. Surgical drainage was performed in 7 patients, with isolation of Staphylococcus aureus in 2 and Pseudomonas aeruginosa in 1. Empirical treatment included 3rd generation cephalosporin, combined in 67% of cases. The median of intravenous antibiotic therapy was 16 days (r: 7-49) with a total of 28 days (r:14-91). One patient required transfer to intensive care unit and 2 nephrectomy. CONCLUSION: Renal abscesses are infrecuent in pediatrics, but they present significant morbidity and mortality. It should be suspected in patients with urinary tract infection (UTI)with torpid evolution that persists with fever without antibiotic response. In our study, the high sensitivity of renal ultrasound allowed early diagnosis.