Identification of the unculturable bacteria Candidatus arthromitus in the intestinal content of trouts using Dot blot and Southern blot techniques

Cecchini, Francesca; Iacumin, Lucilla; Fontanot, Marco; Comi, Giuseppe; Manzano, Marisa

doi:10.1016/j.vetmic.2011.11.020

Cited by 18 publications

(9 citation statements)

References 13 publications

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“…DNA microarrays became a common tool in many areas of microbial research, including detection and identification of bacterial species, due to the high potential to discriminate subtle differences in the genome, the possibility to analyze multiple targets simultaneously, and the ability to detect noncultivable bacteria. In addition, a microarray experiment has the facility to be miniaturized and automated [1, 2, 18]. Thanks to the increase in the complete microbial genome sequencing and the advances in bioinformatics and biomolecular technologies, DNA microarrays are promising analytical tools that are expected to replace time-consuming and laborious conventional techniques for bacterial identification in near future.…”

Section: Discussionmentioning

confidence: 99%

Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes

Böhme

Cremonesi

Villa

et al. 2014

BioMed Research International

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Traditional culturing methods are still commonly applied for bacterial identification in the food control sector, despite being time and labor intensive. Microarray technologies represent an interesting alternative. However, they require higher costs and technical expertise, making them still inappropriate for microbial routine analysis. The present study describes the development of an efficient method for bacterial identification based on flow-through reverse dot-blot (FT-RDB) hybridization on membranes, coupled to the high specific ligation detection reaction (LDR). First, the methodology was optimized by testing different types of ligase enzymes, labeling, and membranes. Furthermore, specific oligonucleotide probes were designed based on the 16S rRNA gene, using the bioinformatic tool Oligonucleotide Retrieving for Molecular Applications (ORMA). Four probes were selected and synthesized, being specific for Aeromonas spp., Pseudomonas spp., Shewanella spp., and Morganella morganii, respectively. For the validation of the probes, 16 reference strains from type culture collections were tested by LDR and FT-RDB hybridization using universal arrays spotted onto membranes. In conclusion, the described methodology could be applied for the rapid, accurate, and cost-effective identification of bacterial species, exhibiting special relevance in food safety and quality.

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Section: Discussionmentioning

confidence: 99%

Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes

Böhme

Cremonesi

Villa

et al. 2014

BioMed Research International

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“…Moreover, specific microorganisms can be detected with probes that anneal to specific DNA sequences [16,10,6,17], and by immunohistochemistry [2].…”

Section: * Corresponding Author Email: Marisamanzano @ Uniudit Inmentioning

confidence: 99%

“…Amplicons obtained from the nested PCR using the RiFCfw and RiFC primers were purified using the QIAquick PCR Purification Kit (Qiagen, Milan, Italy), dried and sent to the MWG sequencing centre (Eurofins MWG GmbH, Martinsried, Germany) for sequencing as reported by Cecchini et al [16].…”

Section: Sequencingmentioning

confidence: 99%

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Utilization of molecular methods to relate Red Mark Syndrome affecting Oncorhynchus mykiss to an unculturable Rickettsiales organism

2017

J App Biol Biotech

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Red Mark Syndrome (RMS) is an inflammatory skin condition related to the presence of an unculturable Rickettsiales organism that affects farmed rainbow trout, Oncorhynchus mykiss. This disease is responsible of economic losses for fish farmers in Europe. The aim of this work was to confirm the presence of Rickettsiales organisms in affected fish and to develop a specific and sensitive system that can detect the unculturable Rickettsiales organisms in farmed trout. Trout gathered from affected and unaffected farms were initially submitted to necropsy and subsequently tissue samples were collected from each fish. Specific primers for a nested PCR (RiFCfw and RiFC), and a DNA probe for dot blot assay were designed using the 16S rRNA sequence. The use of the DNA probe to target amplicons as the template increased the sensitivity to 0.5 pg/µl DNA. Fifteen trout were analysed, of which nine showed visible skin lesions. Eight trout out of the nine with skin lesions were positive for the presence of the unculturable Rickettsiales organism DNA sequence. The tests developed can help rainbow trout producers to quickly treat fish in order to reduce economic losses. Up to 30% of fish with skin lesions are downgraded and rejected at the market by consumers.

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“…However, some microorganisms contained in the water can oxidize and decompose odorous substances, and can effectively remove odor substances. Therefore, it is urgent to find out the sources and causes of odorous substances in water bodies and find a quick and effective odor removal method [3].…”

Section: Introductionmentioning

confidence: 99%

Study on Odor Detection and Microbial Identification Method in Closed Water Area of Jiangxi Province

Yang¹,

Dong²,

Guo³

et al. 2019

I2M

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The problem of odor in water has become one of the most concerned issues of the public. The odor emitted from water will not only lead to deterioration of water quality, but also increase the cost of water treatment, which will have an adverse impact on the ecological environment. This paper takes the Doushui Lake water area as an example to study the odor detection and microbial identification methods by combining Sensory-GC with GC/MS, the research results show that: the water body of the Doushui Lake has a variety of odors, mainly grassy smell, musty smell, earthy smell, pungent smell, liquorice smell and burnt smell. There are four main substances, TCA, Geosmin, β-Cyclocitral and α-Cedrol, and their highest concentrations reach 124 ng/L, 97 ng/L, 43 ng/L and 48 ng/L, respectively. The decomposition of algae by microorganisms produces volatile gas compounds, mainly are dimethyl disulfide and dimethyl tetra sulfide. In the decomposition process, the contents of these two compounds both increase first and decrease later.

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Identification of the unculturable bacteria Candidatus arthromitus in the intestinal content of trouts using Dot blot and Southern blot techniques

Cited by 18 publications

References 13 publications

Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes

Detection of Food Spoilage and Pathogenic Bacteria Based on Ligation Detection Reaction Coupled to Flow-Through Hybridization on Membranes

Utilization of molecular methods to relate Red Mark Syndrome affecting Oncorhynchus mykiss to an unculturable Rickettsiales organism

Study on Odor Detection and Microbial Identification Method in Closed Water Area of Jiangxi Province

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