Infeções em Cateteres de Hemodialisados

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

The fungal microbiota of de-novo paediatric inflammatory bowel disease

Inflammatory bowel disease (IBD) is characterised by an inappropriate chronic immune response against resident gut microbes. This may be on account of distinct changes in the gut microbiota termed as dysbiosis. The role of fungi in this altered luminal environment has been scarcely reported. We studied the fungal microbiome in de-novo paediatric IBD patients utilising next generation sequencing and compared with adult disease and normal controls. We report a distinct difference in fungal species with Ascomycota predominating in control subjects compared to Basidiomycota dominance in children with IBD, which could be as a result of altered tolerance in these patients. 

Characterization of antimicrobial resistance in Aeromonas and Vibrio isolated in Canada from fish and seafood

Plusieurs études ont examiné la sensibilité aux antimicrobiens chez les bactéries d’organismes provenant de produits issus de l’aquaculture ou de leur environnement. Aucune information n’est cependant disponible concernant la résistance aux antimicrobiens dans les bactéries de la flore de poissons ou de fruits de mer vendus au détail au Canada. C’est particulièrement vrai en ce qui a trait aux bactéries des genres Aeromonas et Vibrio, dont certaines espèces sont des agents pathogènes zoonotiques connus. Au cours de cette étude, la sensibilité aux antimicrobiens d’isolats d’Aeromonas spp. et de Vibrio spp. provenant de poissons et de crevettes domestiques et importés a été mesurée à l’aide de techniques de micro dilution en bouillon et/ou de diffusion sur disque. Les classes d’antimicrobiens examinés comprenaient les tétracyclines (TET), les inhibiteurs de la voie des folates (sulfadiméthoxine-triméthoprime, SXT), le florfenicol (FLO), et les quinolones (acide nalidixique / enrofloxacine, NA/ENO). Des valeurs seuils épidémiologiques pour Aeromonas et Vibrio ont été établies en utilisant la méthode d’interprétation normalisée des données de résistance provenant de diffusion sur disque. La recherche de gènes de résistance associés au profil de résistance des isolats a été effectuée en utilisant des PCRs et des puces ADN. Le nombre d’isolats résistants aux divers antimicrobiens parmi les 201 isolats d’Aeromonas et les 185 isolats de Vibrio étaient respectivement les suivants: TET (n=24 et 10), FLO (n=1 et 0), SXT (n=2 et 8), NA (n=7 et 5) et ENO (n= 5 et 0). Diverses associations de gènes tet(A), tet(B), tet(E), floR, sul1, sul2, et intI1 ont été détectées, les gènes tet(E), intI1, sul2 et tet(B) étant les plus communs. Les espèces d’Aeromonas et de Vibrio isolées de poissons au détail et de fruits de mer peuvent héberger une variété de gènes de résistance, bien que peu fréquemment. Le risque que représente ces gènes de résistance reste à évaluer en considérant le potentiel infectieux des bactéries, l’utilisation des ces agents antimicrobiens pour le traitement des maladies en aquaculture et en médecine humaine et leur rôle en tant que réservoir de la résistance antimicrobienne.

Heterotrophic Bacteria Associated with Healthy and Moribund Larvae of Penaeus monodon H.Milne Edwards

Heterotrophic bacterial flora of Pmonadon from an apparently healthy hatchery system as well as a pool with heavy mortality were isolated and studied. In the healthy systems comparatively higher generic diversity with Pseudomonas, Acinetobacter, Bacillus, Micrococcus, members of the family Enterobacteriaceae and coryneform group in the diminishing order of dominance was recorded. Meanwhile from the moribund larvae and rearing water Aeromonas and Pseudomonas could be isolated in almost equal proportions. Strikingly, Aeromonas could not be isolated from the apparently healthy larval rearing system and its exclusive occurrence in the sick culture system in comparatively higher percentage suggested its possible role in the mortality. They were found to be highly halophilic exhibiting growth at 10% NaCl. On testing their sensitivity to twenty antibiotics, four of them (Streptomycin, Gentamycin, Methamine mandelate and Cloramphenicol) were found to be effective on all the isolates of Aeromonas and Pseudomonas suggesting their possible application in the hatchery system in times of emergency. While doing so, Streptomycin would do comparatively better than the others as the minimum inhibitory dose required was comparatively lower (200ppm) within a period of 24 hours

Investigations on Phaeocystis sp., A Harmful Bloom- Forming Alga Isolated from Cochin Estuary

Considering the importance of diversity of micro algae in our ecosystem and new invasion of many organisms, an attempt was made to monitor the Cochin estuary along the south west coast of India for the qualitative distribution of phytoplankton and to study the growth kinetics and allelopathic effect of the phaeocystis sp. Isolated from the cochin estuary. Phaeocystis blooms are common only in high latitude environments and they rarely occur in low latitude environments such as tropics and subtropics. As phaeocystis is grouped under harmful alga ,in the present study the factors causing the blooms formation in the ecosystem. The nutrient concentration of the water body along with other physiochemical parameters that includes temperature salinity and ph play an important role in triggering the bloom of phaeocystis .The phaeocystis harbor specific bacterial flora associated with it and they exert an important role in the growth ,haemolytic activity and the bloom phases of the alga. The harmful alga mainly depends on the production of alleopathic compounds for the establishment of bloom in the marine environments .These physiological properties of the phaecystis were considered for the study, along with the role of nutrients in the allelopathic and hemolytic activity

Studies on Minced Fish Technology

The quality of minced fish, as mentioned earlier depends largely on the type and quality of the raw material used, as well as on the processing methods employed. Moreover, fish mincing involves cutting up of tissues thereby increasing surface area to a great extent and releasing of enzymes and nutrients from the tissues. Due to these factors fish mince is relatively more prone to chemical. autolytic and microbial spoilage. Hence study of minced fish with these factors in focus is very important. Equally important is the availability, price and preference of the raw material vis-a-vis the end products and the storage period it passes through. In the present study. changes in the bacterial flora. both quantitative and qualitative of the dressed fish, viz. Nemipterus japonicas and mince from the same fish during freezing and frozen storage have been investigated in detail. The effect of a preservative. viz. . EZDTA on the bacteriological and shelf life characteristics of the minced fish has also been investigated. Attempts have also been made to develop various types of products from mince and to study their storage life.

Studies on coagulase-positive staphylococci and Vibrio parahaemollticus in selected items of fish, crustaceans and fishery products

In India much work has been done on different aspects of quality of processed fish and fishery products (Pillai et al., 1965; Iyer and Chaudhuri, 1966; Iyer et al., 1966; Sreenivasan and Joseph, 1966; Pillai and Rao, 1969; Mathen et al., 1975; Valsan et al., 1985) and standards have been formulated for almost all the fishery products. Quality standards for fresh fish cover only a few fish species available in the markets. For the formulation and recommendation of quality standards for fish and fishery products for domestic trade detailed background informations are necessary. Khot et al., (1982), Valsan et al. J (1985) and Iyer et al. (1986) have reported on the bacterial flora of sea foods at retail level in Bombay. Cochin is a major fish landing centre of the south west coast of India, and an average of 25,000. tonnes of fish/shell fish are landed annually at this harbour which is about 10 percent of the total catch of fish in Kerala (Lakshmanan -et -al., 1984). All the varieties of marine, brackish water and fresh water fishes are available in Cochln throughout the year for catering to Its cosmopolitan. population and informations regarding their bacteriological quality are scanty. Hence the present study was undertaken to investigate the bacteriological quality of fish and fishery products available in the markets and cold storages situated in and around Cochin meant for internal consumption.

Synechocystis MCCB 114 and 115 as putative probionts for Penaeus monodon post-larvae

Synechocystis MCCB 114 and 115 were segregated as putative probionts for shrimp larvae from a collection of 54 cyanobacterial cultures enriched from seawater. On feeding Penaeus monodon post-larvae with the cyanobacteria, the generic diversity of the intestinal bacterial flora could be enhanced with substantial reduction or total absence of Vibrio spp. A significant difference (p < 0.001) in the percent survival of batches of post-larvae fed on the cyanobacterial cultures was observed and, on repeated challenge with V. harveyi, the relative percent survival of those batches of larvae fed on Synechocystis MCCB 114 and 115 was significantly higher. The Synechocystis MCCB 114 and 115 cultures were found to contain high levels of protein (34 to 43%), in addition to carotenoids

Heterotrophic Bacteria Associated with Healthy and Moribund Larvae of Penaeus monodon H.Milne Edwards

Heterotrophic bacterial flora of Pmonadon from an apparently healthy hatchery system as well as a pool with heavy mortality were isolated and studied. In the healthy systems comparatively higher generic diversity with Pseudomonas, Acinetobacter, Bacillus, Micrococcus, members of the family Enterobacteriaceae and coryneform group in the diminishing order of dominance was recorded. Meanwhile from the moribund larvae and rearing water Aeromonas and Pseudomonas could be isolated in almost equal proportions. Strikingly, Aeromonas could not be isolated from the apparently healthy larval rearing system and its exclusive occurrence in the sick culture system in comparatively higher percentage suggested its possible role in the mortality. They were found to be highly halophilic exhibiting growth at 10% NaCl. On testing their sensitivity to twenty antibiotics, four of them (Streptomycin, Gentamycin, Methamine mandelate and Cloramphenicol) were found to be effective on all the isolates of Aeromonas and Pseudomonas suggesting their possible application in the hatchery system in times of emergency. While doing so, Streptomycin would do comparatively better than the others as the minimum inhibitory dose required was comparatively lower (200ppm) within a period of 24 hours

Diferencias microbiológicas en pacientes con exacerbación severa de EPOC con o sin consolidación neumónica

La EPOC es una causa importante de morbilidad y mortalidad en el mundo y su prevalencia en Bogotá alcanza hasta 8,5%. Las exacerbaciones están asociadas a deterioro funcional y de la calidad de vida por lo que se consideran un factor cardinal de la enfermedad. En la literatura se ha descrito que las infecciones por bacterias y/o virus son las responsables del 78% de las exacerbaciones. Estos datos han sido descritos en poblaciones diferentes y no hay datos en la literatura que muestren cual es la epidemiología local de las exacerbaciones de EPOC y menos aún de aquellas que se asocian a consolidaciones neumónicas. Objetivo: Comparar la microbiología de las exacerbaciones severas de la EPOC que requieren ingreso a UCI con y sin infiltrados alveolares. Materiales y métodos: Estudio de corte transversal en el que se estudiaron pacientes con EPOC que ingresaron a la UCI Médica de la FCI-IC por exacerbación severa, asociada o no a infiltrados alveolares. Se tomaron muestras de microbiología, serológicas y radiografía de tórax para evaluar la etiología de la exacerbación, si se asocia a coinfección viral y a consolidación neumónica o no. Resultados: No se encontró una diferencia estadísticamente significativa en la microbiología de los diferentes grupos evaluados. Se encontró un resistencia global del 24% y llama la atención que hay una alta prevalencia de Serratia Marcescens AMPc entre los 2 grupos, germen que no está descrito como patógeno común en la literatura. Se encontraron diferencias en cuanto a factores de riesgo para presentar neumonía asociada como lo son un mayor índice de paquetes/año (55.1.6 vs. 36.3 paq/año, sig.=0.021). Así mismo se demostró que los pacientes con neumonía asociada presentan mayor necesidad de IOT (48.9 vs. 23.9, sig.=0.013). No hay diferencia significativa en desenlaces como mortalidad (20.5 vs. 13.0, sig.=0.346). Conclusiones: A pesar de no haber diferencia microbiológica entre los 2 grupos se encontraron variables como factores de riesgo y variables clínicas que pueden ayudar a proponer planes de manejo en los dos escenarios. El hecho de encontrar un paciente con neumonía asociada al cuadro de exacerbación no debe afectar en la toma de decisiones en relación al tratamiento antibiótico.

Eficacia de la limpieza corporal con clorhexidina en prevenir la sepsis neonatal en cuidado intensivo revisión sistemática

ANTECEDENTES. La mortalidad neonatal se debe principalmente a procesos infecciosos y a prematurez. Se ha sugerido que el lavado corporal total con clorhexidina podría reducir la mortalidad neonatal relacionada con infección. No existen revisiones sistemáticas que exploren la eficacia de esta intervención. Objetivo. Evaluar la eficacia y seguridad de la limpieza corporal total con clorhexidina en la prevención de las infecciones asociadas al cuidado de la salud en neonatos de alto riesgo hospitalizados en cuidado intensivo neonatal. Metodología. Se realizó una revisión sistemática de la literatura. La búsqueda se hizo a través de las bases de datos Medline, Embase, LilaCS, Cochrane library y el registro de ensayos clínicos del Instituto Nacional de Salud de Estados Unidos. Se incluyeron ensayos clínicos publicados en los últimos 15 años hasta el 30 de enero del 2015. Las variables cualitativas se estimaron mediante OR o RR con sus IC95%. Las variables cuantitativas mediante diferencias de promedios o diferencias estandarizadas de promedios con sus IC95%. Resultados: Se incluyeron 3 estudios en el análisis cualitativo y cuantitativo. No se encontró evidencia concluyente que permita recomendar el uso de la limpieza corporal total con clorhexidina en los recién nacidos hospitalizados en cuidado intensivo neonatal. Conclusión: No existe evidencia que permita concluir que la limpieza corporal total con clorhexidina al 0.25% es mejor respecto a otras intervenciones en la prevención de sepsis neonatal asociada al cuidado de la salud . Es una intervención segura sin efectos adversos significativos.

Molecular characterisation of the gut microflora of healthy and inflammatory bowel disease cats using fluorescence in situ hybridisation with special reference to Desulfovibrio spp

Inflammatory bowel disease (IBD) is a common cause of chronic large bowel diarrhoea in cats. Although the aetiology of IBD is unknown, an immune-mediated response to a luminal antigen is thought to be involved. As knowledge concerning the colonic microflora of cats is limited and requires further investigation, the purpose of this study was to determine the presence of specific bacterial groups in normal and IBD cats, and the potential role they play in the health of the host. Total bacterial populations, Bacteroides spp., Bifidobacterium spp., Clostridium histolyticum subgp., Lactobacillus-Enterococcus subgp. and Desulfovibrio spp. were enumerated in 34 healthy cats and 11 IBD cats using fluorescence in situ hybridisation. The study is one of the first to show the presence of Desulfovibrio in cats. Total bacteria, Bifidobacterium spp. and Bacteroides spp. counts were all significantly higher in healthy cats when compared with IBD cats, whereas Desulfovibrio spp. (producers of toxic sulphides) numbers were found to be significantly higher in colitic cats. The information obtained from this study suggests that modulation of bacterial flora by increasing bifidobacteria and decreasing Desulfovibrio spp. may be beneficial to cats with IBD. Dietary intervention may be an important aspect of their treatment.

The effect of dietary nitrate on salivary, plasma, and urinary nitrate metabolism in humans

Dietary nitrate is metabolized to nitrite by bacterial flora on the posterior surface of the tongue leading to increased salivary nitrite concentrations. In the acidic environment of the stomach, nitrite forms nitrous acid, a potent nitrating/nitrosating agent. The aim of this study was to examine the pharmacokinetics of dietary nitrate in relation to the formation of salivary, plasma, and urinary nitrite and nitrate in healthy subjects. A secondary aim was to determine whether dietary nitrate increases the formation of protein-bound 3-nitrotyrosine in plasma, and if dietary nitrate improves platelet function. The pharmacokinetic profile of urinary nitrate excretion indicates total clearance of consumed nitrate in a 24 h period. While urinary, salivary, and plasma nitrate concentrations increased between 4- and 7-fold, a significant increase in nitrite was only detected in saliva (7-fold). High dietary nitrate consumption does not cause a significant acute change in plasma concentrations of 3-nitrotyrosine or in platelet function.

Prebiotics and lipid metabolism

Prebiotics are defined as nondigestible food ingredients that beneficially affect the host by selectively stimulating the growth or the activity of one or a limited number of bacteria (bifidobacteria, lactobacilli) in the colon. Dietary fructans are nutritionally interesting oligosaccharides that strictly conform to the definition of prebiotics and (in view of experimental studies in animals and of less numerous studies in humans) exhibit interesting serum or hepatic lipid lowering properties. Other nondigestible/fermentable nutrients, which also modulate intestinal flora activity, exhibit cholesterol or triglyceride lowering effects. Are changes in intestinal bacterial flora composition or fermentation activity responsible for those effects? What is the future of prebiotics in the nutritional control of lipidaemia and cardiovascular disease risk in humans? Those questions only receive partial response in the present review because studies of the systemic effects of prebiotics are still in their infancy, and require fundamental research devoted to elucidating the biochemical and physiological events that allow prebiotics to exert systemic effects on lipid metabolism.

The occurrence of treponemes in contagious ovine digital dermatitis and the characterisation of associated Dichelobacter nodosus

Contagious ovine digital dermatitis (CODD) is a recently recorded, apparently new infection of the ovine hoof, which differs clinically from footrot caused by Dichelobacter nodosus and which fails to respond well to accepted treatment practices for footrot. Despite the welfare implications of such an infection, very little research has been performed on CODD to date and the aetiology remains confused. Suggestions have been made that there is a potential role for treponemes in the pathogenesis of CODD but that D. nodosus is apparently not involved. Six farms were therefore targeted in this study to provide a more in-depth investigation into the bacterial flora of CODD lesions. Dark ground microscopy, culture and PCR techniques were used, concentrating on the presence of D. nodosus and spirochaetes, particularly those of the genus Treponema. The results demonstrated that isolates of D. nodosus were indeed present in a high percentage (74%) of CODD lesions compared with 31% of apparently healthy feet. The isolates were shown to be of similar virulence type to those reported previously in cases of footrot, and the range of serogroups was also found to be similar to footrot, with serogroup H being prevalent. Treponemes were present in 70% of CODD lesions and 38% of apparently healthy feet, supporting a possible association between CODD and treponemes. However, any further progress on the aetiology of CODD and the potential for novel, effective treatment will depend on an improved ability to culture these organisms routinely in the laboratory thereby enabling their complete characterisation. (c) 2005 Elsevier B.V. All rights reserved.