A Sco homologue plays a role in defence against oxidative stress in pathogenic Neisseria

Sco proteins are found in mitochondria and in a variety of oxidase positive bacteria. Although Sco is required for the formation of the Cu-A centre in a cytochrome oxidase of the aa(3) type, it was observed that oxidases with a Cu-A centre are not present in many bacteria that contain a Sco homologue. Two bacteria of this type are the pathogens Neisseria meningitidis and Neisseria gonorrhoeae. The sco genes of N. gonorrhoeae strain 1291 and N. meningitidis strain MC58 were cloned, inactivated by inserting a kanamycin resistance cassette and used to make knockout mutants by allelic exchange. Both N. gonorrhoeae and N. meningitidis sco mutants were highly sensitive to oxidative killing by paraquat, indicating that Sco is involved in protection against oxidative stress in these bacteria. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

The genetics of glycosylation in Gram-negative bacteria

In recent years there has been a dramatic increase in reports of glycosylation of proteins in various Gram-negative systems including Neisseria meningitidis, Neisseria gonorrhoeae, Campylobacter jejuni, Pseudomonas aeruginosa, Escherichia coli, Caulobacter crescentus, Aeromonas caviae and Helicobacter pylori. Although this growing list contains many important pathogens (reviewed by Benz and Schmidt [Mol. Microbiol. 45 (2002) 267-276]) and the glycosylations are found on proteins important in pathogenesis such as pili, adhesins and flagella the precise role(s) of the glycosylation of these proteins remains to be determined. Furthermore, the details of the glycosylation biosynthetic process have not been determined in any of these systems. The definition of the precise role of glycosylation and the mechanism of biosynthesis will be facilitated by a detailed understanding of the genes involved. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Ácidos húmicos extraídos do lodo de esgoto sanitário e seus efeitos em plantas

O lodo de esgoto possui alto teor de matéria orgânica porém, também estão presentes diferentes poluentes e patógenos. Desta forma, neste trabalho foi extraído ácido húmico (AH) o qual é um composto resultante do fracionamento de substâncias húmicas que compreendem um grupo de compostos de carbono gerados na decomposição de resíduos orgânicos que sofrem ressíntese formando o húmus. O ácido húmico promove diversos benefícios nos vegetais, como crescimento, elongação celular, tolerância a estresses e aumento da permeabilidade da membrana plasmática. Com base nestas características, o presente trabalho teve como objetivo avaliar os efeitos citogenéticos (ensaio Allium cepa), fisiológicos, anatômicos e bioquímicos do ácido húmico do lodo de esgoto sanitário. Foi realizada caracterização elementar do material para a definição das doses. Após 20 dias de tratamento, foram realizadas coletas do material e, posteriormente, analisadas. A caracterização química do AH indicou-o como bom condicionador para culturas apresentando elevadas taxas de C, H e N. Não foi observado efeito de toxicidade, citotoxicidade, genotoxicidade e mutagenicidade do AH. Foi verificado aumento expressivo de todos os pigmentos fotossintéticos vegetais nas concentrações mais altas (2 mM C L-1 e 4 mM C L-1). Houve aumento da expressão da ATPAse em todos os tratamentos e das enzimas do estresse oxidativo (CAT, SOD, APX, GST) em diferentes concentrações. A integração destas análises permitiu concluir que o ácido húmico do lodo de esgoto pode ser utilizado como adubo orgânico, pois foi observado benefícios no vegetal tais como maior crescimento vegetal e aumento da atividade nas enzimas do estresse oxidativo.

Análise transcriptômica da interação mamoeiro-Papaya Meleira Virus

O mamoeiro (Carica papaya L.) é uma das fruteiras mais cultivadas nas regiões tropicais e subtropicais do mundo. O Brasil faz parte do grupo dos países que mais produzem e exportam mamão no mundo. O Espírito Santo e a Bahia são responsáveis por mais de 70% da área brasileira produtora deste fruto. Porém, doenças causadas por microrganismos infecciosos afetam de modo considerável sua produção. Entre as principais doenças, destaca-se a meleira do mamoeiro, causada pelo Papaya meleira virus (PMeV), que ainda não possui uma cultivar resistente. Interessantemente os sintomas somente são desencadeados após a frutificação. Os mecanismos moleculares envolvidos no desenvolvimento dos sintomas e na resposta de defesa da planta ao PMeV ainda não foram esclarecidos. Para entender os pontos chaves desta interação, que permitam o desenvolvimento de metodologias de melhoramento genético, um estudo transcriptômico foi abordado. A tecnologia RNA-seq foi usada para o sequenciamento do transcriptoma a partir de plantas com 3, 6 e 8 meses de idade após plantio, inoculadas e não inoculadas com o PMeV. Os genes diferencialmente expressos nos 3 tempos e nas duas condições foram preditos e analisados. Estas análises revelaram um padrão de expressão geral dos genes envolvidos nesta interação. Foram encontrados 21 genes com o perfil de expressão alterado nas plantas inoculadas exclusivamente nos seis meses de idade. Destes, 8 genes envolvidos em processos de respostas de defesa e morte celular, resposta ao estresse e resposta ao estímulo biótico e abiótico foram reprimidos; enquanto os demais (13 genes), envolvidos principalmente em processos metabólicos primários, biogêneses, diferenciação e ciclo celular, comunicação e crescimento celular, bem como processos envolvidos em reprodução, e desenvolvimento da floração, foram superexpressos. Estes resultados sugerem que, aos seis meses de idade, a planta é obrigada a alterar seu programa de expressão gênica, direcionando a resposta para os processos próprios do desenvolvimento, requeridos nesse estádio fisiológico, que primam sob a resposta ao estresse, fato que finalmente leva ao desenvolvimento dos sintomas.

Uniformity of liquid distribution in the canopy of the bean plant, using the spectrophotometric analysis

Fungal diseases are important factors limiting common bean yield. White mold is one of the main diseases caused by soil pathogens. The objective of this study was to quantify the distribution of a fungicide solution sprayed into the canopy of bean plants by spectrophotometry, using a boom sprayer with and without air assistance. The experiment was arranged in a 2 x 2 x 2 factorial (two types of nozzles, two application rates, and air assistance on and off) randomized block design with four replications. Air assistance influenced the deposition of solution on the bean plant and yield increased significantly with the increased rate of application and air assistance in the boom sprayer.

Rootstocks resistant to Meloidogyne incognita and compatibility of grafting in net melon

Due to the few studies about grafting in net melon, in order to obtain better control of soil pathogens, the aim of the present study was to evaluate 16 genotypes of Cucurbitaceae: Benincasa hispida, Luffa cylindrica, pumpkin 'Jacarezinho', pumpkin 'Menina Brasileira', squash 'Exposição', squash 'Coroa', pumpkin 'Canhão Seca', pumpkin 'Squash', pumpkin 'Enrrugado Verde', pumpkin 'Mini Paulista', pumpkin 'Goianinha', watermelon 'Charleston Gray', melon 'Rendondo Gaucho', melon 'Redondo Amarelo', cucumber 'Caipira HS' and cucumber 'Caipira Rubi', regarding to compatibility of grafting in net melon and resistance to Meloidogyne incognita, based on the reproduction factor (RF), according to Oostenbrink (1966). To assess resistance, the seedlings were transplanted to ceramic pots and inoculated with 300/mL eggs and/or second stage juveniles of M. incognita. At 50 days after transplanting, the plants were removed from the pots and the resistance was evaluated. The compatibility between resistant rootstock and grafts of net melon was determined by performing simple cleft grafting, in a commercial net melon hybrid of great market acceptance and susceptible to M. incognita (Bonus no. 2). The genotypes Luffa cylindrica, pumpkin 'Goianinha', pumpkin 'Mini-Paulista', melon 'Redondo Amarelo', watermelon 'Charleston Gray' are resistant to the nematode M. incognita. The better compatibilities occurred with the rootstocks melon 'Amarelo', which presented 100% of success, followed by pumpkin 'Mini-Paulista' with 94%. On the other hand, Sponge gourd, watermelon 'Charleston Gray' and pumpkin 'Goianinha' showed low graft take percentages of 66%, 62% and 50%, respectively.

Heat shock and salicylic acid on postharvest preservation of organic strawberries

Heat shock and salicylic acid have been studied on shelf-life extension of fruits. The benefits of these techniques have been related to their effect on inducing physiological defense responses against the oxidative stress and pathogen development. The objective of this study was to evaluate the effect of heat shock and salicylic acid on the postharvest preservation and contents of total phenolics, anthocyanins, ascorbic acid, fresh weight loss and microbiological quality of organic strawberries cv. Dover. Strawberries produced organically and stored at 5 ºC were subjected to heat shock (45 ºC ± 3 ºC for 3 h), application of salicylic acid (soaking in 2.0 mmol L-1 solution), heat shock in combination with salicylic acid and control. After treatment, the fruits were packed and stored in a climatic chamber at 5 ºC ± 2 ºC. At 1, 7 and 14 days, the experimental units were removed from refrigeration and kept at room temperature of approximately 20 ºC for two days. There was no effect of treatments on fresh weight loss, incidence of pathogens or chemical variations in strawberry fruits during the storage period. In natural conditions, organically grown strawberries remained in good condition for sale up to seven days of storage in all treatments.

Application of GIS-based multi-criteria analysis for site selection of aquifer recharge with reclaimed water

Reclaimed water from small wastewater treatment facilities in the rural areas of the Beira Interior region (Portugal) may constitute an alternative water source for aquifer recharge. A 21-month monitoring period in a constructed wetland treatment system has shown that 21,500 m(3) year(-1) of treated wastewater (reclaimed water) could be used for aquifer recharge. A GIS-based multi-criteria analysis was performed, combining ten thematic maps and economic, environmental and technical criteria, in order to produce a suitability map for the location of sites for reclaimed water infiltration. The areas chosen for aquifer recharge with infiltration basins are mainly composed of anthrosol with more than 1 m deep and fine sand texture, which allows an average infiltration velocity of up to 1 m d(-1). These characteristics will provide a final polishing treatment of the reclaimed water after infiltration (soil aquifer treatment (SAT)), suitable for the removal of the residual load (trace organics, nutrients, heavy metals and pathogens). The risk of groundwater contamination is low since the water table in the anthrosol areas ranges from 10 m to 50 m. Oil the other hand, these depths allow a guaranteed unsaturated area suitable for SAT. An area of 13,944 ha was selected for study, but only 1607 ha are suitable for reclaimed water infiltration. Approximately 1280 m(2) were considered enough to set up 4 infiltration basins to work in flooding and drying cycles.

Environmental impact on the bacteriological quality of domestic water supplies in Lagos, Nigeria

OBJECTIVE: To assess the impact of town planning, infrastructure, sanitation and rainfall on the bacteriological quality of domestic water supplies. METHODS: Water samples obtained from deep and shallow wells, boreholes and public taps were cultured to determine the most probable number of Escherichia coli and total coliform using the multiple tube technique. Presence of enteric pathogens was detected using selective and differential media. Samples were collected during both periods of heavy and low rainfall and from municipalities that are unique with respect to infrastructure planning, town planning and sanitation. RESULTS: Contamination of treated and pipe distributed water was related with distance of the collection point from a utility station. Faults in pipelines increased the rate of contamination (p<0.5) and this occurred mostly in densely populated areas with dilapidated infrastructure. Wastewater from drains was the main source of contamination of pipe-borne water. Shallow wells were more contaminated than deep wells and boreholes and contamination was higher during period of heavy rainfall (p<0.05). E. coli and enteric pathogens were isolated from contaminated supplies. CONCLUSIONS: Poor town planning, dilapidated infrastructure and indiscriminate siting of wells and boreholes contributed to the low bacteriological quality of domestic water supplies. Rainfall accentuated the impact.

Diversidade e prospecção de fungos endofíticos de Begonia spp. encontradas na Mata Atlântica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Role on serine protease inhibitors in the pathogenisis of Steinernema carpocapsae

Tese de Doutoramento, Biologia (Biologia Celular e Molecular), 18 de Novembro de 2013, Universidade dos Açores.

Routine screening of harmful microorganisms in beach sands: implications to public health

Beaches worldwide provide recreational opportunities to hundreds of millions of people and serve as important components of coastal economies. Beach water is often monitored for microbiological quality to detect the presence of indicators of human sewage contamination so as to prevent public health outbreaks associated with water contact. However, growing evidence suggests that beach sand can harbor microbes harmful to human health, often in concentrations greater than the beach water. Currently, there are no standards for monitoring, sampling, analyzing, or managing beach sand quality. In addition to indicator microbes, growing evidence has identified pathogenic bacteria, viruses, and fungi in a variety of beach sands worldwide. The public health threat associated with these populations through direct and indirect contact is unknown because so little research has been conducted relating to health outcomes associated with sand quality. In this manuscript, we present the consensus findings of a workshop of experts convened in Lisbon, Portugal to discuss the current state of knowledge on beach sand microbiological quality and to develop suggestions for standardizing the evaluation of sand at coastal beaches. The expert group at the "Microareias 2012" workshop recommends that 1) beach sand should be screened for a variety of pathogens harmful to human health, and sand monitoring should then be initiated alongside regular water monitoring; 2) sampling and analysis protocols should be standardized to allow proper comparisons among beach locations; and 3) further studies are needed to estimate human health risk with exposure to contaminated beach sand. Much of the manuscript is focused on research specific to Portugal, but similar results have been found elsewhere, and the findings have worldwide implications.

Diversity and characterisation of coagulasenegative staphylococci isolated from healthy individuals in Portugal

In the past few years the interest in coagulase-negative staphylococci (CoNS) has significantly increased in human medicine. CoNS are common commensal colonisers of the human skin, although now also recognised as major nosocomial pathogens. Over the last decades, several studies have been carried out in order to understand the pathogenicity mechanisms of CoNS. The well known determinants in the pathogenesis of CoNS infections are their ability to form biofilms and an exceptional resistance to several antibiotics. Nevertheless, there is a lack of studies regarding the commensal lifestyle of these microorganisms. Additionally, it is now hypothesised that commensal bacteria might be a reservoir of pathogenic determinants. Therefore, the work described throughout this thesis was aimed to perform a phenotypic and genotypic characterisation of different CoNS species isolated from healthy Portuguese individuals. A total of 61 CoNS isolates, comprising 7 different species, were obtained and characterised at the level of biofilm formation and antibiotic susceptibility profiles. According to the results, biofilm formation ability and presence of biofilm-associated genes were commonly found features, highlighting their pivotal role in the colonising lifestyle of CoNS. This study also addressed the correlation between phenotypic and genotypic characteristics of biofilm formation, corroborating and raising questions about the importance of some genes in this process. Moreover, it was observed a great proportion of isolates with decreased susceptibility and multiple resistances to some important antibiotics. A significant association between antibiotic resistance and biofilm formation was also demonstrated, and some hypotheses about the nature of such association were provided. Lastly, the expression patterns of two biofilm-associated genes at two distinct biofilm developmental stages were determined, confirming their importance in the accumulative stage of biofilm formation. Overall, the results presented in this thesis indicate that staphylococcal skin flora might be an important reservoir of potentially pathogenic bacteria and, simultaneously, bring to light new perceptions about the molecular basis of staphylococcal biofilm formation, and the nature of the association between antibiotic resistance and biofilm formation.

Immune responses in Bathymodiolus azoricus upon Vibrio challenges : approaches to characterize mussel survival strategies and physiological adaptations in deep sea extreme environments

Tese de Doutoramento, Ciências do Mar, especialidade de Biologia Marinha, 18 de Dezembro de 2015, Universidade dos Açores.

Beach sand and the potential for infectious disease transmission: observations and recommendations

Recent studies suggest that sand can serve as a vehicle for exposure of humans to pathogens at beach sites, resulting in increased health risks. Sampling for microorganisms in sand should therefore be considered for inclusion in regulatory programmes aimed at protecting recreational beach users from infectious disease. Here, we review the literature on pathogen levels in beach sand, and their potential for affecting human health. In an effort to provide specific recommendations for sand sampling programmes, we outline published guidelines for beach monitoring programmes, which are currently focused exclusively on measuring microbial levels in water. We also provide background on spatial distribution and temporal characteristics of microbes in sand, as these factors influence sampling programmes. First steps toward establishing a sand sampling programme include identifying appropriate beach sites and use of initial sanitary assessments to refine site selection. A tiered approach is recommended for monitoring. This approach would include the analysis of samples from many sites for faecal indicator organisms and other conventional analytes, while testing for specific pathogens and unconventional indicators is reserved for high-risk sites. Given the diversity of microbes found in sand, studies are urgently needed to identify the most significant aetiological agent of disease and to relate microbial measurements in sand to human health risk.