Bacterial diseases in marine bivalves

Bivalve aquaculture is seriously affected by many bacterial pathogens that cause high losses in hatcheries as well as in natural beds. A number of Vibrio species, but also members of the genera Nocardia and Roseovarius, are considered important pathogens in aquaculture. The present work provides an updated overview of main diseases and implicated bacterial species affecting bivalves. This review focuses on aetiological agents, their diversity and virulence factors, the diagnostic methods available as well as information on the dynamics of the host-parasite relationship.

Combining ability of Phaseolus vulgaris L. for resistance to common bacterial blight

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Specific detection of the cleavage activity of mycobacterial enzymes using a quantum dot based DNA nanosensor

We present a quantum dot based DNA nanosensor specifically targeting the cleavage step in the reaction cycle of the essential DNA-modifying enzyme, mycobacterial topoisomerase I. The design takes advantages of the unique photophysical properties of quantum dots to generate visible fluorescence recovery upon specific cleavage by mycobacterial topoisomerase I. This report, for the first time, demonstrates the possibility to quantify the cleavage activity of the mycobacterial enzyme without the pre-processing sample purification or post-processing signal amplification. The cleavage induced signal response has also proven reliable in biological matrices, such as whole cell extracts prepared from Escherichia coli and human Caco-2 cells. It is expected that the assay may contribute to the clinical diagnostics of bacterial diseases, as well as the evaluation of treatment outcomes.

Health status of a snakehead (Channa punctatus) of two fish markets in Mymensingh, Bangladesh

An experiment was carried out for a period of six months during October 2008 to March 2009 to investigate the health status of a snakehead, Channa punctatus through clinical and histopathological technique. Fish were collected from two fish markets of Mymensingh district. Clinically and histopathologically, it was observed that fishes from both the markets were healthy in October and March but moderately affected in November and February. In the months of December and January, 7.5- 8% of the fishes were affected clinically and showing various clinical signs like, discolouration, deep ulcer, ill health, scale loss and rough skin. Histopathologically, in the month of December and January, major observed pathologies of skin and muscle were necrosis, vacuums, fungal granuloma and loss of dermis. Gills were affected having parasitic cysts, monogenetic trematode, clubbing, loss of primary and secondary gill lamellae, hemorrhage, necrosis and hypertrophy. Vacuoles, pyknosis, hepatic necrosis, hemorrhages and fungal granuloma were observed in liver. Renal pathology included necrosis and pyknosis of kidney tubules, hemorrhages, presence of bacterial colony and vacuoles. From present findings, it was found that, fishes from urban market were more affected with diseases than pre-urban market especially in the months of December and January when compared with other months. From overall observation, C. punctatus were severely affected by epizootic ulcerative syndrome (EUS), dactylogyrosis, protozoan and bacterial diseases during colder months of the year.

Diversity among bacteria causing blotch disease on the commercial mushroom, agaricus bisporus /

A total of 251 bacterial isolates were isolated from blotched mushroom samples obtained from various mushroom farms in Canada. Out of 251 stored isolates, 170 isolates were tested for pathogenicity on Agaricus bisporus through mushroom rapid pitting test with three distinct pathotypes observed: dark brown, brovm and yellow/yellow-brown blotch. Phenotypic analysis of 83 isolates showed two distinct proteinase K resistant peptide profiles. Profile group A isolates exhibited peptides with masses of 45, 18, 16 and 14 kDa and fiirther biochemical tests identified them as Pseudomonasfluorescens III and V. Profile group B isolates lacked the 16-kDa peptide and the blotch causing bacterial isolates of this group was identified as Serratia liquefaciens and Cedecea davisae. Comparative genetic analysis using Amplified Fragment Length Polymorphism (AFLP) on 50 Pseudomonas sp. isolates (Group A) showed that various blotch symptoms were caused by isolates distributed throughout the Pseudomonas sp. clusters with the exception of the Pseudomonas tolaasii group and one non-pathogenic Pseudomonas fluorescens cluster. These results show that seven distinct Pseudomonas sp. genotypes (genetic clusters) have the ability to cause various symptoms of blotch and that AFLP can discriminate blotch causing from non-blotch causing Pseudomonasfluorescens. Therefore, a complex of diverse bacterial organisms causes bacterial blotch disease

Cell wall degrading enzymes and interaction between Trichoderma Aggressivum and Agaricus Bisporus

Agaricus bisporus is the most commonly cultivated mushroom in North America and has a great economic value. Green mould is a serious disease of A. bisporus and causes major reductions in mushroom crop production. The causative agent of green mould disease in North America was identified as Trichoderma aggressivum f. aggressivum. Variations in the disease resistance have been shown in the different commercial mushroom strains. The purpose of this study is to continue investigations of the interactions between T. aggressivum and A. bisporus during the development of green mould disease. The main focus of the research was to study the roles of cell wall degrading enzymes in green mould disease resistance and pathogenesis. First, we tried to isolate and sequence the N-acetylglucosaminidase from A. bisporus to understand the defensive mechanism of mushroom against the disease. However, the lack of genomic and proteomic information of A. bisporus limited our efforts. Next, T. aggressivum cell wall degrading enzymes that are thought to attack Agaricus and mediate the disease development were examined. The three cell wall degrading enzymes genes, encoding endochitinase (ech42), glucanase (fJ-1,3 glucanase) and protease (prb 1), were isolated and sequenced from T. aggressivum f. aggressivum. The sequence data showed significant homology with the corresponding genes from other fungi including Trichoderma species. The transcription levels of the three T. aggressivum cell wall degrading enzymes were studied during the in vitro co-cultivation with A. bisporus using R T -qPCR. The transcription levels of the three genes were significantly upregulated compared to the solitary culture levels but were upregulated to a lesser extent in co-cultivation with a resistant strain of A. bisporus than with a sensitive strain. An Agrobacterium tumefaciens transformation system was developed for T. aggressivum and was used to transform three silencing plasmids to construct three new T. aggressivum phenotypes, each with a silenced cell wall degrading enzyme. The silencing efficiency was determined by RT-qPCR during the individual in vitro cocultivation of each of the new phenotypes with A. bisporus. The results showed that the expression of the three enzymes was significantly decreased during the in vitro cocultivation when compared with the wild type. The phenotypes were co-cultivated with A. bisporus on compost with monitoring the green mould disease progression. The data indicated that prbi and ech42 genes is more important in disease progression than the p- 1,3 glucanase gene. Finally, the present study emphasises the role of the three cell wall degrading enzymes in green mould disease infection and may provide a promising tool for disease management.

Protein subcellular targeting inTrichoderma aggressivum f. aggressivum

Trichoderma aggressivum f. aggressivum is a filamentous soil fungus. Green mold disease of commercial mushrooms caused by this species in North America has resulted in millions of dollars in lost revenue within the mushroom growing industry. Research on the molecular level of T aggressivum have jus t begun with the goal of understanding the functions of each gene and protein, and their expression control. Protein targeting has not been well studied in this species yet. Therefore, the intent of this study was to test the protein localization and production levels in T aggressivum with green fluorescent protein (GFP) with an intron and tagged with either nuclear localization signal (NLS) or an endoplasmic reticulum retention signal (KDEL). Two GFP constructs (with and without the intron) were used as controls in this study. All four constructs were successfully transferred into T aggressivum and all modified strains showed similar growth characteristics as the wild type non-transformed isolate. GFP expression was detected from all modified T aggressivum with confocal microscopy and the expression was similar in all four strains. The intron tested in this study had no or very minor effects as GFP expression was similar with or without it. The GFP signal increased over a 5 day period for all transformants, while the GFP to total protein ratio decreased over the same period for all transformants. The GFP-KDEL transformant showed similar protein expression level and localization as did the control transformant lacking the KDEL retention signal. The GFP-NLS transformant similarly failed to localize GFP into nucleus as fluorescence with this strain was virtually identical to the GFP transformant lacking the NLS. Thus, future research is required to find effective localization signals for T aggressivum.

Inhibition du pathogène des salmonidés Saprolegnia parasitica par des bactéries aquatiques

Les maladies constituent présentement la cause la plus importante de perte économique en aquaculture moderne. Chez certaines espèces, notamment les salmonidés (Oncorhynchus sp. et Salmo sp.), on rapporte des pertes annuelles atteignant cinquante pour cent de la production. À l’heure actuelle, les infections fongiques occupent le second rang derrière les maladies bactériennes en fonction de leur importance économique. Ces poissons sont particulièrement vulnérables à une infection fongique causée par Saprolegnia sp. qui infecte habituellement les oeufs morts. Le saprophyte ubiquitaire se propage ensuite aux oeufs sains et aux individus matures. Malheureusement, le traitement efficace de cette infection, souvent primaire et parfois secondaire, est de plus en plus difficile en raison de nouvelles réglementations restrictives entourant le vert de malachite. Jadis, ce colorant constituait le fongicide le plus efficace dans la lutte contre la saprolégniose, mais son potentiel cancérigène en limite maintenant l’utilisation. Jusqu'à présent, aucun traitement disponible n’est aussi efficace que le vert de malachite pour le contrôle de la saprolégniose. Récemment, nous sommes parvenus à isoler trois bactéries capables d’inhiber la croissance de Saprolegnia sp. in vitro. Ces trois Pseudomonas fluorescens proviennent d’une pisciculture dans laquelle survenaient des cas d’infections à Saprolegnia parasitica. En poussant la caractérisation de l’activité grâce à des analyses de chromatographie liquide haute performance et de spectrométrie de masse, nous avons réussi à isoler et à identifier la molécule responsable. L’acide phénazine-1-carboxylique (PCA), sécrété par deux de nos trois souches, cause l’inhibition de la croissance de Saprolegnia.

Two isoforms of anti-lipopolysaccharide factors identified and characterized from the hemocytes of portunid crabs, Portunus pelagicus and Scylla tranquebarica

Anti-lipopolysaccharide factors (ALFs), a type of cationic antimicrobial peptides (AMPs), and their derivatives are becoming predominant candidates for potential drugs in viral and bacterial diseases. This study reports the first ALF from the mud crab Scylla tranquebarica (StALF, JQ899453) and the second ALF isoform from the blue swimmer crab Portunus pelagicus (PpALF2, JQ899452). Both sequences encoded for precursor molecules, starting with a signal peptide containing 26 amino acid residues, followed by a highly cationic mature peptide, containing two conserved cysteine residues flanking a putative lipopolysaccharide (LPS)-binding domain. BLAST analysis revealed that both PpALF2 and StALF exhibited significant similarity with crustacean ALF sequences. The predicted molecular mass of the mature ALFs was 11.2 kDa with an estimated pI of 10.0. PpALF2 and StALF also showed the typical pattern of alternating hydrophobic and hydrophilic residues in their putative disulphide loop, suggesting that they comprise the same functional domain. Phylogenetic analysis showed that PpALF2 and StALF have similar evolutionary status and they were phylogenetically ancient immune effector molecules which may play an essential role in the host defense mechanism. The spatial structures of PpALF2 and StALF possessed four beta-strands and two alpha-helices. The results indicated that there were more than one ALF involved in crab immunity against various pathogens. ALFs would provide candidate promising therapeutic or prophylactic agents in health management and diseases control in crustacean aquaculture

Fotoperíodo, temperatura e reguladores vegetais na germinação de sementes de Passiflora cincinnata Mast.

A espécie Passiflora cincinnata Mast., pertencente à família Passifloraceae, é silvestre e popularmente conhecida como maracujá-do-mato, sendo considerada importante na produção de porta-enxertos, uma vez que é tolerante à seca, a doenças causadas por bactérias e a nematóides, além de poder ser utilizada em programas de melhoramento genético. O trabalho teve como objetivos estudar o efeito da luz e da temperatura e a interação entre temperatura e reguladores vegetais na germinação de sementes de Passiflora cincinnata Mast. Foi constituído de três experimentos: no primeiro, estudou-se o efeito da luz e da temperatura na germinação de sementes; no segundo, o efeito de diferentes concentrações dos reguladores vegetais GA4+7 + N-(fenilmetil)-aminopurina na germinação das sementes e, no terceiro, a interação entre temperatura e reguladores vegetais na germinação das sementes. O delineamento experimental foi inteiramente casualizado para todos os experimentos e os dados foram submetidos à análise de variância e comparação das médias pelo teste Tukey, a 5% de probabilidade. É possível observar que a luz exerce efeito inibitório sobre a germinação das sementes, e que os reguladores vegetais, GA4+7 + N-(fenilmetil)-aminopurina, são eficientes na superação da dormência, além de ampliarem os limites de temperatura da germinação. A temperatura alternada 20-30ºC mostra-se a mais adequada para a germinação de sementes dessa espécie.

Efectos del parasitismo sobre el tejido branquial en peces cultivados: estudio parasitologico e histopatologico

One hundred four out of 225 diagnosticated cases were myxosporidian, monogenean, Ichthyophthirius multifiliis Fouquet, 1876 and bacterial diseases in Piaractus mesopotamicus Holmberg, 1887 (pacu), Colossoma macropomum Cuvier, 1818 (tambaqui) and tambacu, at Aquaculture Center, Universidade Estadual Paulista (UNESP), Jaboticabal, São Paulo, Brazil, between 1992 and 1995. The gills were fixed in 10% buffered formalin solution to posterior histologic routine. It was observed Henneguya sp. cysts into the capillaries of the secondary lamellae, encapsulated by the respiratory epithelium cells. It provoked adherence of the adjacent lamellae, hyperplasia, congestion, oedema and epithelium displacement. Monogeneans infestations by Anacanthorus penilabiatus Boeger, Husak & Martins, 1995 and Ancyrocephalinae sub-family caused light inflammatory reaction and hyperplasia. In severe infestations was observed hyperplasia of primary lamellae, necrosis, oedema, respiratory epithelium displacement, ruptured pillar cells and telangiectasis. Response to I. multifiliis was limited to surrounding epithelial cells in young fishes and hyperplasia, necrosis, inflammatory infiltrate and oedema in old fishes. Such lesions iniciate hyperplasic and oedematous process that with inflammation of the parasitic sites, provoked alterations over gases and ions interchange surface and consequently fish metabolism.

Lipossomas: Estrategia biotecnologica para liberacao controlada e direcionamento intracelular de farmacos com efeito antimicobacteriano

This text highlights the state of research related with the application of liposomes in the control of drug delivery and drug target to intracellular bacterial diseases, such as the tuberculosis. Liposome have several pharmaceutical applications and this article is primarily focused on the potential of this agregate on drug encapsalation especially antimycobacterial compounds. Case studies in which liposomes have successfully been used to improve pharmacological drug effect are presented. Mechanisms involved in intracellular drug delivery, possibilities of application, research and development efforts to address these objectives are discussed.

Ecotoxicidade e eficácia da oxitetraciclina e do florfenicol contra infecção experimental por Aeromonas hydrophila e aspectos histopatológicos em pacu (Piaractus mesopotamicus)

Pós-graduação em Aquicultura - FCAV

Curtobacterium flaccumfaciens PV. flaccumfaciens: sobrevivência, gama de hospedeiras e efeito do pré-plantio de aveia e trigo na ocorrência da doença

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)